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Volume 20,Issue 2,2013 Table of Contents
2013, 20(2):129-137.
DOI: 10.3872/j.issn.1007-385X.2013.02.001
Abstract:
In recent years, the clinical trials on using cytokine induced killer (CIK) cells for solid tumor therapy have been launched worldwide. CIK cell immunotherapy has showed great advantages in clinical treatment compared with other adoptive immunity therapy, which is therefore more and more widely used in adoptive immunotherapy of tumors. CIK cells can be used alone or in combination with surgery, radiotherapy and chemotherapy for comprehensive treatment. In this article, we keep a track of research progress both home and abroad, briefly introduce the source, phenotype, activation and amplification of the CIK cells, and summarize their anti-tumor mechanisms, new techniques and methods, and current status of international clinical trials from several aspects. Some open questions about clinical treatment, standard management, quality monitoring, therapeutic evaluation and future directions are also explored.
In recent years, the clinical trials on using cytokine induced killer (CIK) cells for solid tumor therapy have been launched worldwide. CIK cell immunotherapy has showed great advantages in clinical treatment compared with other adoptive immunity therapy, which is therefore more and more widely used in adoptive immunotherapy of tumors. CIK cells can be used alone or in combination with surgery, radiotherapy and chemotherapy for comprehensive treatment. In this article, we keep a track of research progress both home and abroad, briefly introduce the source, phenotype, activation and amplification of the CIK cells, and summarize their anti-tumor mechanisms, new techniques and methods, and current status of international clinical trials from several aspects. Some open questions about clinical treatment, standard management, quality monitoring, therapeutic evaluation and future directions are also explored.
2013, 20(2):138-144.
DOI: 10.3872/j.issn.1007-385X.2013.02.002
Abstract:
Objective:To investigate the feasibility of targeted fluorescent imaging to MGC-803 gastric cancer cells and the potential of CT imaging of gastric cancer using nanocomposite probes of silica-coated gold nanocluster (AuNC@SiO2). Methods: The AuNC was coated by multilayer silica shells to obtain AuNC@SiO2. AuNC@SiO2 was conjugated with cancer cell targeted folic acid (FA), and AuNC@SiO2-FA was then used to investigate the feasibility of targeted fluorescent imaging to gastric cancer cells. MG-803 cells were injected subcutaneously into BABL/c nude mice to establish a cancer xenograft model. CT imaging was performed to collect CT signals before and after subcutaneously injection of AuNC@SiO2, thus exploring the feasibility of AuNC@SiO2 nanoparticles in CT imaging of tumor mouse models. Results: AuNC@SiO2 was successfully constructed with a uniform shape, good dispersbility and fluorescence optical properties. AuNC@SiO2 and AuNC@SiO2-FA showed a low cytotoxicity on gastric cancer MGC-803 cells and normal gastric mucosal GES-1 cells at a concentration of 500 mg/ml. The AuNC@SiO2-FA showed an effectively targeted ability to gastric cancer MGC-803 cells rather than normal GES-1 cells, thus presenting a clearly fluorescence imaging. CT signals were significantly enhanced in tumors after AuNC@SiO2 injection, with HU value increasing from 129.16 before injection to 383.32. Conclusion: AuNC@SiO2 can be used for fluorescence imaging and CT dual mode imaging. Moreover, AuNC@SiO2 conjugated with FA can realize gastric cancer cells targeted fluorescence imaging.
Objective:To investigate the feasibility of targeted fluorescent imaging to MGC-803 gastric cancer cells and the potential of CT imaging of gastric cancer using nanocomposite probes of silica-coated gold nanocluster (AuNC@SiO2). Methods: The AuNC was coated by multilayer silica shells to obtain AuNC@SiO2. AuNC@SiO2 was conjugated with cancer cell targeted folic acid (FA), and AuNC@SiO2-FA was then used to investigate the feasibility of targeted fluorescent imaging to gastric cancer cells. MG-803 cells were injected subcutaneously into BABL/c nude mice to establish a cancer xenograft model. CT imaging was performed to collect CT signals before and after subcutaneously injection of AuNC@SiO2, thus exploring the feasibility of AuNC@SiO2 nanoparticles in CT imaging of tumor mouse models. Results: AuNC@SiO2 was successfully constructed with a uniform shape, good dispersbility and fluorescence optical properties. AuNC@SiO2 and AuNC@SiO2-FA showed a low cytotoxicity on gastric cancer MGC-803 cells and normal gastric mucosal GES-1 cells at a concentration of 500 mg/ml. The AuNC@SiO2-FA showed an effectively targeted ability to gastric cancer MGC-803 cells rather than normal GES-1 cells, thus presenting a clearly fluorescence imaging. CT signals were significantly enhanced in tumors after AuNC@SiO2 injection, with HU value increasing from 129.16 before injection to 383.32. Conclusion: AuNC@SiO2 can be used for fluorescence imaging and CT dual mode imaging. Moreover, AuNC@SiO2 conjugated with FA can realize gastric cancer cells targeted fluorescence imaging.
2013, 20(2):145-152.
DOI: 10.3872/j.issn.1007-385X.2013.02.003
Abstract:
Objective: To investigate the targeted migration and killing effect of mesenchymal stem cells (MSCs) infected with Lenti-TK (thymidine kinase) vector on CD133+ cancer stem cells in nasopharyngeal carcinoma. Methods: The recombinant lentiviral expression vector containing TK gene (Lenti-TK) was constructed and transduced into MSC (Lenti-TK-MSC). Fusion tag-TK (HA-TK) expression was verified by RT-PCR and Western blotting. CD133+ cells were sorted from nasopharyngeal carcinoma 5-8F cells by immunomagnetic beads. The chemotactic ability of Lenti-TK-MSC to CD133+5-8F cells was analyzed by Transwell assay. The CD133+ 5-8F cells were co-cultured with Lenti-TK-MSC and GCV to detect its killing effect on cells by CCK-8 Kit. Results: The recombinant lentivirus vector Lenti-TK was successfully constructed with titer being 1×108 UT/ml. The transduction efficiency of Lenti-TK to MSC was (95.1±0.1)%, 72 h after transduction at an MOI of 50. The migration number of Lenti-TK-MSC to CD133+5-8F cells was more than that to CD133-5-8F cells and 5-8F cells (\[83.0±8.7\] vs \[29.6±5.3\] vs \[38.3±5.2\]；P=0.000). The Lenti-TK-MSC/GCV treatment significantly inhibited the growth of the CD133+ 5-8F cells compared with the GCV group and the Lenti-TK-MSC/GCV condition medium group (the culture supernatant of Lenti-TK-MSC treated with 1 mg/L GCV for 48 h) (\[37.2±2.3\]% vs \[98.5±3.1\]% vs \[83.8±3.4\]%, P=0.000), with the cell survival being (68.2±2.3)% when the proportion of Lenti-TK-MSC was 20%, which showed a significant bystander killing effect. Conclusion: Lenti-TK infected MSC exert targeted migration and killing effects on CD133+ 5-8F cells from nasopharyngeal carcinoma.
Objective: To investigate the targeted migration and killing effect of mesenchymal stem cells (MSCs) infected with Lenti-TK (thymidine kinase) vector on CD133+ cancer stem cells in nasopharyngeal carcinoma. Methods: The recombinant lentiviral expression vector containing TK gene (Lenti-TK) was constructed and transduced into MSC (Lenti-TK-MSC). Fusion tag-TK (HA-TK) expression was verified by RT-PCR and Western blotting. CD133+ cells were sorted from nasopharyngeal carcinoma 5-8F cells by immunomagnetic beads. The chemotactic ability of Lenti-TK-MSC to CD133+5-8F cells was analyzed by Transwell assay. The CD133+ 5-8F cells were co-cultured with Lenti-TK-MSC and GCV to detect its killing effect on cells by CCK-8 Kit. Results: The recombinant lentivirus vector Lenti-TK was successfully constructed with titer being 1×108 UT/ml. The transduction efficiency of Lenti-TK to MSC was (95.1±0.1)%, 72 h after transduction at an MOI of 50. The migration number of Lenti-TK-MSC to CD133+5-8F cells was more than that to CD133-5-8F cells and 5-8F cells (\[83.0±8.7\] vs \[29.6±5.3\] vs \[38.3±5.2\]；P=0.000). The Lenti-TK-MSC/GCV treatment significantly inhibited the growth of the CD133+ 5-8F cells compared with the GCV group and the Lenti-TK-MSC/GCV condition medium group (the culture supernatant of Lenti-TK-MSC treated with 1 mg/L GCV for 48 h) (\[37.2±2.3\]% vs \[98.5±3.1\]% vs \[83.8±3.4\]%, P=0.000), with the cell survival being (68.2±2.3)% when the proportion of Lenti-TK-MSC was 20%, which showed a significant bystander killing effect. Conclusion: Lenti-TK infected MSC exert targeted migration and killing effects on CD133+ 5-8F cells from nasopharyngeal carcinoma.
2013, 20(2):153-158.
DOI: 10.3872/j.issn.1007-385X.2013.02.004
Abstract:
Objective: To investigate the association between the expression of the ten-eleven translocation 2 (TET2) protein and the cytotoxin associated gene A (CagA) of Helicobacter pylori (Hp)，and to explore the possible mechanisms of CagA in the process of gastric carcinogenesis. Methods: Real-time PCR was used to detect TET2 mRNA level in human gastric epithelial GES-1 cells and gastric cancer MGC-803 cells. Immuncytochemistry was used to detect the TET2 protein localization and expression in GES-1 and MGC-803 cells. GES-1 cells were transfected with pEGFP-CagA and a cell oxidative stress model was constructed with 200 μmol/L H2O2. Flow cytometry was used to analyze cell cycle and ROS in GES-1 cells. Results: The mean expression level of TET2 mRNA in GES-1 cells was lower than that in MGC-803 cells (1.00±0.08 vs 1.68±0.07, P<0.05). TET2 protein expression in GES-1 cells was lower than that in MGC-803 cells (8.09±3.57 vs 14.60±2.31，P<0.05). Compared with the negative control pEGFP-N1 group, the mean expression level of TET2 mRNA in pEGFP-CagA-transfected GES-1 cells was increased (1.00±0.04 vs 0.06±000，P<005), and TET2 protein expression in pEGFP-CagA-transfected GES-1 cells was also increased (10.82±3.39 vs 16.45±4.40，P<0.05), and a higher level of ROS production was observed in pEGFP-CagA-transfected GES-1 cells (69±90 vs 91±16.8，P<0.05), which significantly displayed the cell apoptosis in cycle analysis. In the cell oxidative stress model, TET2 mRNA level in H2O2 treated cells was higher than that in normal GES-1 cells (1.44±0.02 vs 1.00±0.04，P<005) and TET2 protein expression level was higher than that in normal GES-1 cells (11.74±4.34 vs 15.72±452，P<0.05). Conclusion: CagA factor can induce ROS accumulation and cell cycle disruption of GES-1, indicating that TET2 can be upregulated by oxidative stress and may be involved in the progress of CagA-induced carcinogenesis.
Objective: To investigate the association between the expression of the ten-eleven translocation 2 (TET2) protein and the cytotoxin associated gene A (CagA) of Helicobacter pylori (Hp)，and to explore the possible mechanisms of CagA in the process of gastric carcinogenesis. Methods: Real-time PCR was used to detect TET2 mRNA level in human gastric epithelial GES-1 cells and gastric cancer MGC-803 cells. Immuncytochemistry was used to detect the TET2 protein localization and expression in GES-1 and MGC-803 cells. GES-1 cells were transfected with pEGFP-CagA and a cell oxidative stress model was constructed with 200 μmol/L H2O2. Flow cytometry was used to analyze cell cycle and ROS in GES-1 cells. Results: The mean expression level of TET2 mRNA in GES-1 cells was lower than that in MGC-803 cells (1.00±0.08 vs 1.68±0.07, P<0.05). TET2 protein expression in GES-1 cells was lower than that in MGC-803 cells (8.09±3.57 vs 14.60±2.31，P<0.05). Compared with the negative control pEGFP-N1 group, the mean expression level of TET2 mRNA in pEGFP-CagA-transfected GES-1 cells was increased (1.00±0.04 vs 0.06±000，P<005), and TET2 protein expression in pEGFP-CagA-transfected GES-1 cells was also increased (10.82±3.39 vs 16.45±4.40，P<0.05), and a higher level of ROS production was observed in pEGFP-CagA-transfected GES-1 cells (69±90 vs 91±16.8，P<0.05), which significantly displayed the cell apoptosis in cycle analysis. In the cell oxidative stress model, TET2 mRNA level in H2O2 treated cells was higher than that in normal GES-1 cells (1.44±0.02 vs 1.00±0.04，P<005) and TET2 protein expression level was higher than that in normal GES-1 cells (11.74±4.34 vs 15.72±452，P<0.05). Conclusion: CagA factor can induce ROS accumulation and cell cycle disruption of GES-1, indicating that TET2 can be upregulated by oxidative stress and may be involved in the progress of CagA-induced carcinogenesis.
Wang Ke
,
Wang Yi
,
Liu Guijun
,
Han Gencheng
,
Wang Renxi
,
Xiao He
,
Hou Chunmei
,
Pen Hui
,
Shen Beifen
,
Li Yan
,
Chen Guojiang
2013, 20(2):159-165.
DOI: 10.3872/j.issn.1007-385X.2013.02.005
Abstract:
Objective: To investigate the role and mechanism of vascular endothelial growth factor (VEGF) signaling in the development of ulcerative colitis (UC)-related cancer. Methods: Colitis-associated colorectal cancer (CAC) model was established in Balb/c mice. The proportion of myeloid-derived suppressor cells (MDSCs) in the peripheral blood, spleen, bone marrow and tumor tissues of the model mice and lesions was examined by flow cytometry. Arginase-1 (Arg-1) mRNA and inducible nitric oxide synthase (iNOS) mRNA expressions in MDSCs were detected by qPCR. The VEGF expression in the supernatants of colonic tissues was determined by ELISA. Sorafenib or neutralizing anti-VEGF antibody was used to block VEGF signaling and the proportion of MDSC in colonic lesions and the histopathology of CAC were detected. Results: Murine CAC model was established successfully. 1 month and 3 months after the beginning of CAC were verified as the early and late stages of CAC respectively, according to several parameters. The increased number of Gr-1+CD11b+MDSC was observed in the progression of CAC and was more significant in lesions: control (0.30±018)%, early stage of CAC (1.32±0.04)%, late stage of CAC (3.08±0.29)% (P<0.05). These MDSCs expressed a high level of Arg-1 and iNOS (P<0.05). But the levels of L-arginine in colonic lesions of CAC mice was much lower than controls (early stage of CAC \[4.22±0.17\] μg/ml, late stage of CAC \[2.95±1.08\] μg/ml, control \[4.41±0.16\] μg/ml, P<0.05). Furthermore, VEGF expression in the lesions of CAC mice was elevated significantly (early stage of CAC \[1170±94.43\] pg/ml, late stage of \[1117±71.92\] pg/ml, control \[877.6±31.67\] pg/ml, P<0.05). The treatment of sorafenib or anti-VEGF dramatically reduced accumulation of MDSC in the lesions. Conclusion: VEGF plays a pro-tumor role in CAC formation, which may be related with the induced accumulation of MDSC in colonic tissues.
Objective: To investigate the role and mechanism of vascular endothelial growth factor (VEGF) signaling in the development of ulcerative colitis (UC)-related cancer. Methods: Colitis-associated colorectal cancer (CAC) model was established in Balb/c mice. The proportion of myeloid-derived suppressor cells (MDSCs) in the peripheral blood, spleen, bone marrow and tumor tissues of the model mice and lesions was examined by flow cytometry. Arginase-1 (Arg-1) mRNA and inducible nitric oxide synthase (iNOS) mRNA expressions in MDSCs were detected by qPCR. The VEGF expression in the supernatants of colonic tissues was determined by ELISA. Sorafenib or neutralizing anti-VEGF antibody was used to block VEGF signaling and the proportion of MDSC in colonic lesions and the histopathology of CAC were detected. Results: Murine CAC model was established successfully. 1 month and 3 months after the beginning of CAC were verified as the early and late stages of CAC respectively, according to several parameters. The increased number of Gr-1+CD11b+MDSC was observed in the progression of CAC and was more significant in lesions: control (0.30±018)%, early stage of CAC (1.32±0.04)%, late stage of CAC (3.08±0.29)% (P<0.05). These MDSCs expressed a high level of Arg-1 and iNOS (P<0.05). But the levels of L-arginine in colonic lesions of CAC mice was much lower than controls (early stage of CAC \[4.22±0.17\] μg/ml, late stage of CAC \[2.95±1.08\] μg/ml, control \[4.41±0.16\] μg/ml, P<0.05). Furthermore, VEGF expression in the lesions of CAC mice was elevated significantly (early stage of CAC \[1170±94.43\] pg/ml, late stage of \[1117±71.92\] pg/ml, control \[877.6±31.67\] pg/ml, P<0.05). The treatment of sorafenib or anti-VEGF dramatically reduced accumulation of MDSC in the lesions. Conclusion: VEGF plays a pro-tumor role in CAC formation, which may be related with the induced accumulation of MDSC in colonic tissues.
6 Ad-ING4 combined with radiotherapy suppresses growth of SPC-A1 cell-transplanted lung adenocarcinoma
2013, 20(2):166-171.
DOI: 10.3872/j.issn.1007-385X.2013.02.006
Abstract:
Objective: To study the anticancer effect of adenovirus-mediated inhibitor of growth family 4 (Ad-ING4) combined with radiotherapy on SPC-A1 transplanted lung adenocarcinoma. Methods: SPC-A1 tumor-bearing mice were established and anticancer experiment was conducted on these mice after being randomly divided into 5 groups: PBS group, Ad group, Ad-ING4 group, radiotherapy group and Ad-ING4 combined with radiotherapy group. Tumor growth was recorded by volume. All tumors were taken out and weighed at 15 days after treatment. The morphology of lung adenocarcinoma SPC-A1 cells was observed by H-E staining. The expressions of Bax, caspase-3, Bcl-2, and VEGF were detected by immunohistochemisty. Results: The tumor volume of the Ad-ING4 combined with radiotherapy group was much smaller than that of the Ad-ING4 group and the single radiotherapy group (\[743.84±109.06\]mm3 vs \[1136.03±151.58\]mm3, \[1035.67±86.27\]mm3, P<0.01). The anticancer rate of the Ad-ING4 combined with radiotherapy group was also much better than that of the Ad-ING4 group and the single radiotherapy group (69.62% vs 33.17% vs 35.41%, P<001), which showed an enhanced radiosensitivity and a synergetic effect (Q=1.22). Immunohistochemistry results showed that Ad-ING4 combined with radiotherapy could obviously up-regulate the expressions of Bax, caspase-3 and down-regulate the expressions of Bcl-2 and VEGF. And the effect of the combined group was better than the single Ad-ING4 or radiotherapy group (P<0.01). Conclusion: Ad-ING4 combined with radiotherapy can effectively inhibit the growth of SPC-A1 cell-transplanted lung adenocarcinoma.
Objective: To study the anticancer effect of adenovirus-mediated inhibitor of growth family 4 (Ad-ING4) combined with radiotherapy on SPC-A1 transplanted lung adenocarcinoma. Methods: SPC-A1 tumor-bearing mice were established and anticancer experiment was conducted on these mice after being randomly divided into 5 groups: PBS group, Ad group, Ad-ING4 group, radiotherapy group and Ad-ING4 combined with radiotherapy group. Tumor growth was recorded by volume. All tumors were taken out and weighed at 15 days after treatment. The morphology of lung adenocarcinoma SPC-A1 cells was observed by H-E staining. The expressions of Bax, caspase-3, Bcl-2, and VEGF were detected by immunohistochemisty. Results: The tumor volume of the Ad-ING4 combined with radiotherapy group was much smaller than that of the Ad-ING4 group and the single radiotherapy group (\[743.84±109.06\]mm3 vs \[1136.03±151.58\]mm3, \[1035.67±86.27\]mm3, P<0.01). The anticancer rate of the Ad-ING4 combined with radiotherapy group was also much better than that of the Ad-ING4 group and the single radiotherapy group (69.62% vs 33.17% vs 35.41%, P<001), which showed an enhanced radiosensitivity and a synergetic effect (Q=1.22). Immunohistochemistry results showed that Ad-ING4 combined with radiotherapy could obviously up-regulate the expressions of Bax, caspase-3 and down-regulate the expressions of Bcl-2 and VEGF. And the effect of the combined group was better than the single Ad-ING4 or radiotherapy group (P<0.01). Conclusion: Ad-ING4 combined with radiotherapy can effectively inhibit the growth of SPC-A1 cell-transplanted lung adenocarcinoma.
7 Cytotoxicity effect of parthenolide on cancer stem cells in mouse breast cancer stem cells in vivo
Wang Xinrong
,
Tian Lianfang
,
Sun Chunxiu
,
Li Yuehong
,
Zhang Haipu
,
Li Shujun
,
Liu Lihua
,
Shan Baoen
2013, 20(2):172-176.
DOI: 10.3872/j.issn.1007-385X.2013.02.007
Abstract:
Objective:To investigate the effects of parthenolide (PTL) on cancer stem cells (CSCs) of mouse breast cancer in vivo, providing an experimental basis for the clinical treatment of breast cancer with PTL. Methods: The mouse breast cancer model which enriching for 4T1 breast cancer CSCs was established by chemotherapeutic drug 5-fluorouracil (5-FU). Tumor-bearing mice were randomly divided into three groups, a control group, a 5-FU group and a PTL group. Four weeks later, the mice were sacrificed. The mouse tumor volume and weight were measured. Flow cytometry (FCM) was used to measure the proportion of CD44+CD24-/low cells in different tumor tissues; Hoechst 33342 staining was used to measure the proportion of SP (side population) cells; and the expressions of ALDH1 (aldehyde dehydrogenase1) and CD55 proteins were detected by immunohistochemistry. The formation of mammosphere was observed under an inverted microscope. Results: The mouse breast cancer transplanted model enriching for 4T1 breast cancer CSCs was successfully prepared. PTL could significantly reduce the proportion of CD44+CD24-/low cells in the mouse transplanted tumor tissues （\[42.5±3.7\]% vs \[687±3.2\]%，P<0.05), decrease the percentage of the SP cells in mouse transplanted tumor tissues （\[39.2±18\]% vs \[61.3±2.6\]%，P<0.05), and inhibit the expressions of CD55 and ALDH1 proteins in the mouse tumor tissues （\[18.9±1.5\]% vs \[30.1±1.3\]%，\[8.1±2.3\]% vs \[18.0±1.4\]%; all P<0.05). PTL inhibited the formation of mammosphere generated from the mouse tumor 4T1 cells culturing in serum-free medium and the growth of the mouse transplanted tumor (\[0.625±0.159\]cm3 vs \[1.715±0.184\] cm3, \[1.467±0.373\]g vs \[3.367±0.398\] g; P<0.05\]. Conclusion: PTL can significantly reduce the CSC contents in mouse tumor tissues in vivo, which suggests that PTL could be used to target the breast cancer CSC.
Objective:To investigate the effects of parthenolide (PTL) on cancer stem cells (CSCs) of mouse breast cancer in vivo, providing an experimental basis for the clinical treatment of breast cancer with PTL. Methods: The mouse breast cancer model which enriching for 4T1 breast cancer CSCs was established by chemotherapeutic drug 5-fluorouracil (5-FU). Tumor-bearing mice were randomly divided into three groups, a control group, a 5-FU group and a PTL group. Four weeks later, the mice were sacrificed. The mouse tumor volume and weight were measured. Flow cytometry (FCM) was used to measure the proportion of CD44+CD24-/low cells in different tumor tissues; Hoechst 33342 staining was used to measure the proportion of SP (side population) cells; and the expressions of ALDH1 (aldehyde dehydrogenase1) and CD55 proteins were detected by immunohistochemistry. The formation of mammosphere was observed under an inverted microscope. Results: The mouse breast cancer transplanted model enriching for 4T1 breast cancer CSCs was successfully prepared. PTL could significantly reduce the proportion of CD44+CD24-/low cells in the mouse transplanted tumor tissues （\[42.5±3.7\]% vs \[687±3.2\]%，P<0.05), decrease the percentage of the SP cells in mouse transplanted tumor tissues （\[39.2±18\]% vs \[61.3±2.6\]%，P<0.05), and inhibit the expressions of CD55 and ALDH1 proteins in the mouse tumor tissues （\[18.9±1.5\]% vs \[30.1±1.3\]%，\[8.1±2.3\]% vs \[18.0±1.4\]%; all P<0.05). PTL inhibited the formation of mammosphere generated from the mouse tumor 4T1 cells culturing in serum-free medium and the growth of the mouse transplanted tumor (\[0.625±0.159\]cm3 vs \[1.715±0.184\] cm3, \[1.467±0.373\]g vs \[3.367±0.398\] g; P<0.05\]. Conclusion: PTL can significantly reduce the CSC contents in mouse tumor tissues in vivo, which suggests that PTL could be used to target the breast cancer CSC.
2013, 20(2):177-181.
DOI: 10.3872/j.issn.1007-385X.2013.02.008
Abstract:
Objective: To explore the effect of the down-regulation of Med19 expression by RNAi on the proliferation and apoptosis of colon cancer Caco-2 cells. Methods: The pSilencer-Med19-siRNA interference plasmid targeting Med19 was constructed, which was transfected into Caco-2 cells. The level of Med19 mRNA in transfected Caco-2 cells was detected by RT-PCR and the level of Med19 protein was determined by Western blotting. Flow cytometry and MTT were performed to detect the proliferation and apoptosis of Caco-2 cells after pSilencer-Med19-siRNA transfection. Results: The results of RT-PCR and Western blotting showed that the mRNA and protein levels of Med19 declined markedly in Caco-2 cells that were transfected by pSilencer-Med19-siRNA vector (P<0.01). Compared with the control pSilencer group, flow cytometry and MTT assay demonstrated that pSilencer-Med19-siRNA significantly suppressed proliferation （7 d: \[086±0.09\]% vs \[1.38±1.10\]%, P<0.01) and induced the apoptosis of Caco-2 cells (\[22.72±2.85\]% vs \[723±1.29\]%, P<0.01) in the pSilencer-Med19-siRNA group. Conclusion: Silence Med19 expression by interference RNA can inhibit the proliferation and induce apoptosis of colon cancer Caco-2 cells, so Med19 may act as a potential therapeutic target in colon cancer.
Objective: To explore the effect of the down-regulation of Med19 expression by RNAi on the proliferation and apoptosis of colon cancer Caco-2 cells. Methods: The pSilencer-Med19-siRNA interference plasmid targeting Med19 was constructed, which was transfected into Caco-2 cells. The level of Med19 mRNA in transfected Caco-2 cells was detected by RT-PCR and the level of Med19 protein was determined by Western blotting. Flow cytometry and MTT were performed to detect the proliferation and apoptosis of Caco-2 cells after pSilencer-Med19-siRNA transfection. Results: The results of RT-PCR and Western blotting showed that the mRNA and protein levels of Med19 declined markedly in Caco-2 cells that were transfected by pSilencer-Med19-siRNA vector (P<0.01). Compared with the control pSilencer group, flow cytometry and MTT assay demonstrated that pSilencer-Med19-siRNA significantly suppressed proliferation （7 d: \[086±0.09\]% vs \[1.38±1.10\]%, P<0.01) and induced the apoptosis of Caco-2 cells (\[22.72±2.85\]% vs \[723±1.29\]%, P<0.01) in the pSilencer-Med19-siRNA group. Conclusion: Silence Med19 expression by interference RNA can inhibit the proliferation and induce apoptosis of colon cancer Caco-2 cells, so Med19 may act as a potential therapeutic target in colon cancer.
Chu Huili
,
Wang Jun
,
Zhu Zhongpeng
,
Guo Yan
,
Wang Baocheng
,
Bi Jingwang
,
Li Kainan
,
Liang Xiuju
2013, 20(2):182-186.
DOI: 10.3872/j.issn.1007-385X.2013.02.009
Abstract:
Objective: To investigate the effect of interference of human antigen R (HuR) expression on sensitivity of human multidrug-resistant human breast cancer MCF-7/Adr cell line to Doxorubicn. Methods: The shRNA expression vector targeting HuR gene (pGenesil-siHuR) has been constructed and stably transfected into human breast cancer MCF-7/Adr cell line. The expression level of MDR1 mRNA in MCF-7/Adr cells was assayed by real-time PCR. The P-gp protein (encoded by the MDR1 gene) expression were determined by Western blotting. The survival rate and IC50 of MCF-7/Adr cells to doxorubicin after pGenesil-siHuR transfection were evaluated by MTT method. The apoptosis rate of MCF-7/Adr cells was detected by flow cytometry. Results: Compared with untransfected MCF-7/Adr cells, the MDR1 mRNA（\[0.184±0.029\] vs \[1.203±0.026\]，P<0.01）and P-gp protein expressions（\[0.314±0.011\] vs \[0.796±0007\], P<0.01）were significantly reduced in pGenesil-siHuR transfected MCF-7/Adr cells (P<0.01). The IC50 of MCF-7/Adr cells to doxorubicin decreased from (148.2±2.3) nmol/L to (42.9±0.4) nmol/L after pGenesil-siHuR transfection. Compared with untransfected MCF-7/Adr cells, the ratio of cell apoptosis was significantly increased in pGenesil-siHuR transfected MCF-7/Adr cells （\[34.6±1.1\]% vs \[1.1±0.2\]%, P<0.01）after the treatment with doxorubicin. Conclusion: RNA interference of HuR can inhibit the expression of MDR1 gene and increase the sensitivity of multidrug-resistant breast cancer cells to doxorubicin.
Objective: To investigate the effect of interference of human antigen R (HuR) expression on sensitivity of human multidrug-resistant human breast cancer MCF-7/Adr cell line to Doxorubicn. Methods: The shRNA expression vector targeting HuR gene (pGenesil-siHuR) has been constructed and stably transfected into human breast cancer MCF-7/Adr cell line. The expression level of MDR1 mRNA in MCF-7/Adr cells was assayed by real-time PCR. The P-gp protein (encoded by the MDR1 gene) expression were determined by Western blotting. The survival rate and IC50 of MCF-7/Adr cells to doxorubicin after pGenesil-siHuR transfection were evaluated by MTT method. The apoptosis rate of MCF-7/Adr cells was detected by flow cytometry. Results: Compared with untransfected MCF-7/Adr cells, the MDR1 mRNA（\[0.184±0.029\] vs \[1.203±0.026\]，P<0.01）and P-gp protein expressions（\[0.314±0.011\] vs \[0.796±0007\], P<0.01）were significantly reduced in pGenesil-siHuR transfected MCF-7/Adr cells (P<0.01). The IC50 of MCF-7/Adr cells to doxorubicin decreased from (148.2±2.3) nmol/L to (42.9±0.4) nmol/L after pGenesil-siHuR transfection. Compared with untransfected MCF-7/Adr cells, the ratio of cell apoptosis was significantly increased in pGenesil-siHuR transfected MCF-7/Adr cells （\[34.6±1.1\]% vs \[1.1±0.2\]%, P<0.01）after the treatment with doxorubicin. Conclusion: RNA interference of HuR can inhibit the expression of MDR1 gene and increase the sensitivity of multidrug-resistant breast cancer cells to doxorubicin.
2013, 20(2):187-191.
DOI: 10.3872/j.issn.1007-385X.2013.02.010
Abstract:
Objective: To explore the effect of the hepatic stellate cell condition medium (HSC-CM) on chemo-resistance of human hepatocellular carcinoma PLC/PRF/5 cells and its possible mechanism. Methods: Hepatic stellate cell line LX-2 was incubated and activated in serum-free RPMI 1640 medium, and then this condition medium was collected, named HSC-CM. PLC/PRF/5 cells were incubated in HSC-CM for 24 h. After the treatment of cisplatin for 12 or 24 h, the apoptosis of PLC/PRF/5 cells was identified by flow cytometry, the cell proliferation of PLC/PRF/5 cells was detected by MTT assay, and the expression of epithelial mesenchymal transition (EMT) associated genes in PLC/PRF/5 cells was detected by real-time PCR. Results: The apoptosis rates of PLC/PRF/5 cells in the cisplatin group were (2234±112)% and (26.78±1.56)%; those in the HSC-CM+cisplatin group were (17.22±1.42)% and (21.52±176)% at 12 and 24 h time points, which showed that the apoptosis rates of the cisplatin group were higher than of the HSC-CM+cisplatin group (P<0.05). The proliferation of PLC/PRF/5 cells in the cisplatin group and HSC-CM+cisplatin group at these two time points were (68.65±2.56)% and （79.47±1.43)%, (46.54±3.65)% and (62.77±289) % respectively, showing a stronger proliferation activity of PLC/PRF/5 cells from the HSC-CM+cisplatin group. Real-time PCR results indicated that compared with the cisplatin group, the expression of epithelial marker E-cadherin in PLC/PRF/5 cells from the HSC-CM+cisplatin group was decreased (P<0.05), and the mesenchymal markers (N-cadherin, vimentin) and EMT-associated transcription factor (Snail, ZEB1) expressions were significantly up-regulated (P<0.01). Conclusion: HSC-CM may promote the rchemo-resistance of PLC/PRF/5 cells through inducing EMT of PLC/PRF/5 cells.
Objective: To explore the effect of the hepatic stellate cell condition medium (HSC-CM) on chemo-resistance of human hepatocellular carcinoma PLC/PRF/5 cells and its possible mechanism. Methods: Hepatic stellate cell line LX-2 was incubated and activated in serum-free RPMI 1640 medium, and then this condition medium was collected, named HSC-CM. PLC/PRF/5 cells were incubated in HSC-CM for 24 h. After the treatment of cisplatin for 12 or 24 h, the apoptosis of PLC/PRF/5 cells was identified by flow cytometry, the cell proliferation of PLC/PRF/5 cells was detected by MTT assay, and the expression of epithelial mesenchymal transition (EMT) associated genes in PLC/PRF/5 cells was detected by real-time PCR. Results: The apoptosis rates of PLC/PRF/5 cells in the cisplatin group were (2234±112)% and (26.78±1.56)%; those in the HSC-CM+cisplatin group were (17.22±1.42)% and (21.52±176)% at 12 and 24 h time points, which showed that the apoptosis rates of the cisplatin group were higher than of the HSC-CM+cisplatin group (P<0.05). The proliferation of PLC/PRF/5 cells in the cisplatin group and HSC-CM+cisplatin group at these two time points were (68.65±2.56)% and （79.47±1.43)%, (46.54±3.65)% and (62.77±289) % respectively, showing a stronger proliferation activity of PLC/PRF/5 cells from the HSC-CM+cisplatin group. Real-time PCR results indicated that compared with the cisplatin group, the expression of epithelial marker E-cadherin in PLC/PRF/5 cells from the HSC-CM+cisplatin group was decreased (P<0.05), and the mesenchymal markers (N-cadherin, vimentin) and EMT-associated transcription factor (Snail, ZEB1) expressions were significantly up-regulated (P<0.01). Conclusion: HSC-CM may promote the rchemo-resistance of PLC/PRF/5 cells through inducing EMT of PLC/PRF/5 cells.
2013, 20(2):192-196.
DOI: 10.3872/j.issn.1007-385X.2013.02.011
Abstract:
Objective: To evaluate the effect of recombinant human adenovirus- P53 (rAd-P53) combined with paclitaxel on proliferation, apoptosis and vascular endothelial growth factor (VEGF) expression of cervical cancer HeLa cells. Methods: The effects of paclitaxel, rAd-P53 alone or in combination on the proliferation of HeLa cells were evaluated by MTT. The effects of paclitaxel, rAd-P53 alone or in combination on apoptosis of HeLa cells at 48 h were evaluated by DAPI stain. The effects of paclitaxel, rAd-P53 alone or in combination on VEGF expression in HeLa cells were examined by Western blotting. Results: Paclitaxel, rAd-P53 alone or in combination inhibited HeLa cell proliferation significantly at 24-72 h in dose-dependent and time-dependent manners. Furthermore, the inhibitory effect was more significant with rAd-P53 combined with paclitaxel than with paclitaxel or rAd-P53 alone group (P<0.05). The coefficients of drug interaction (CDI) of various doses of rAd-P53 combined with paclitaxel were less than 1, which indicated that there was a synergism between them. At 48 h of treatment, rAd-P53 (5×107 VP/ml) combined with paclitaxel (3 μg/ml) showed stronger cell inhibition than the two drugs treated alone (\[54.0 ± 0.92\]% vs \[31.8 ± 0.58\]%, \[27.2±0.55\]%, P<0.05). In addition, the combined group demonstrated more powerful ability in apoptosis induction than paclitaxel and rAd-P53 alone（ \[83±0.07\]% vs \[36±0.04\]%,\[62±0.05\]%，P<0.05), but the expression of VEGF in HeLa cells in the combined treatment group decreased more significantly than did the single drug groups (\[81 ± 0.08\]% vs \[45±0.07\]%, \[60±0.06\]%, P<0.05). Conclusion: rAd-P53 combined with paclitaxel shows more significant proliferation inhibition and apoptosis induction effects than does paclitaxel or rAd-P53 alone. Its mechanism may be associated with down-regulation of VEGF expression.
Objective: To evaluate the effect of recombinant human adenovirus- P53 (rAd-P53) combined with paclitaxel on proliferation, apoptosis and vascular endothelial growth factor (VEGF) expression of cervical cancer HeLa cells. Methods: The effects of paclitaxel, rAd-P53 alone or in combination on the proliferation of HeLa cells were evaluated by MTT. The effects of paclitaxel, rAd-P53 alone or in combination on apoptosis of HeLa cells at 48 h were evaluated by DAPI stain. The effects of paclitaxel, rAd-P53 alone or in combination on VEGF expression in HeLa cells were examined by Western blotting. Results: Paclitaxel, rAd-P53 alone or in combination inhibited HeLa cell proliferation significantly at 24-72 h in dose-dependent and time-dependent manners. Furthermore, the inhibitory effect was more significant with rAd-P53 combined with paclitaxel than with paclitaxel or rAd-P53 alone group (P<0.05). The coefficients of drug interaction (CDI) of various doses of rAd-P53 combined with paclitaxel were less than 1, which indicated that there was a synergism between them. At 48 h of treatment, rAd-P53 (5×107 VP/ml) combined with paclitaxel (3 μg/ml) showed stronger cell inhibition than the two drugs treated alone (\[54.0 ± 0.92\]% vs \[31.8 ± 0.58\]%, \[27.2±0.55\]%, P<0.05). In addition, the combined group demonstrated more powerful ability in apoptosis induction than paclitaxel and rAd-P53 alone（ \[83±0.07\]% vs \[36±0.04\]%,\[62±0.05\]%，P<0.05), but the expression of VEGF in HeLa cells in the combined treatment group decreased more significantly than did the single drug groups (\[81 ± 0.08\]% vs \[45±0.07\]%, \[60±0.06\]%, P<0.05). Conclusion: rAd-P53 combined with paclitaxel shows more significant proliferation inhibition and apoptosis induction effects than does paclitaxel or rAd-P53 alone. Its mechanism may be associated with down-regulation of VEGF expression.
2013, 20(2):197-200.
DOI: 10.3872/j.issn.1007-385X.2013.02.012
Abstract:
Objective: To explore the effects of IL-12 on the phenotypes of cytokine-induced killer (CIK) cells and the cytolytic activity of CIK cells against human esophageal carcinoma EC9706 cells in vitro. Methods: Peripheral blood mononuclear cells were isolated from healthy donors. Two groups were designed: a control group (cells were cultured in the presence of IFN-γ, IL-2 and anti-CD3 antibody) and IL-12 group (cells were cultured in the presence of IFN-γ, anti-CD3 antibody , IL-2 and IL-12 ). After 14-day culture, the phenotypes of CIK cells in the control and IL-12 groups were analyzed by flow cytometry. The cytotoxic activity of CIK cells on EC9706 cells was measured by LDH releasing assay at effect-to-target (E ∶T) cell ratios of 20 ∶1, 30 ∶1. The anti-NKG2D monoclonal antibody was added to CIK cells to detect its effect on the cytotoxic activity of CIK cells at E ∶T ratio of 30 ∶1. Results: In comparison to the control group, the proportion of CD3+CD56+cells, NKG2D expressions on CD3+ and CD3+CD56+cells, and perforin expression in CD3+ cells were higher in the IL-12 group (\[28.23±1.71\]% vs \[16.34±0.59\]%; \[77.45±2.15\]% vs \[66.87±0.73\]%,\[92.94±0.77\]% vs \[82.18±0.66\]%; \[51.78±0.63\]% vs \[43.54±095\]%; all P<0.05). Whereas, no significant change was observed in the granzyme B expression in the CD3+ cells \[(26.90±0.67）% vs (26.76±033）%，P>0.05\]. The cytolytic activity of CIK cells against EC9706 cells was increased significantly in the IL-12 group (E ∶T ratio of 20 ∶1, \[43.92± 1.67\]% vs \[35.34±1.22\]% ; E ∶T ratio of 30 ∶1, \[55.95±0.88\]% vs \[43.91±1.10\]%, all P<0.05). The cytotoxicity of CIK cells in the IL-12 and the control groups were significantly inhibited by anti-NKG2D monoclonal antibody (\[19.72±0.56\]% vs \[55.95±0.88\]%, \[19.83±1.20\]% vs \[43.91±1.10\]%, all P<0.05). Conclusion: IL-12 up-regulates the NKG2D and perforin expressions on CIK cells, enhancing their cytotoxicity against EC9706 cells.
Objective: To explore the effects of IL-12 on the phenotypes of cytokine-induced killer (CIK) cells and the cytolytic activity of CIK cells against human esophageal carcinoma EC9706 cells in vitro. Methods: Peripheral blood mononuclear cells were isolated from healthy donors. Two groups were designed: a control group (cells were cultured in the presence of IFN-γ, IL-2 and anti-CD3 antibody) and IL-12 group (cells were cultured in the presence of IFN-γ, anti-CD3 antibody , IL-2 and IL-12 ). After 14-day culture, the phenotypes of CIK cells in the control and IL-12 groups were analyzed by flow cytometry. The cytotoxic activity of CIK cells on EC9706 cells was measured by LDH releasing assay at effect-to-target (E ∶T) cell ratios of 20 ∶1, 30 ∶1. The anti-NKG2D monoclonal antibody was added to CIK cells to detect its effect on the cytotoxic activity of CIK cells at E ∶T ratio of 30 ∶1. Results: In comparison to the control group, the proportion of CD3+CD56+cells, NKG2D expressions on CD3+ and CD3+CD56+cells, and perforin expression in CD3+ cells were higher in the IL-12 group (\[28.23±1.71\]% vs \[16.34±0.59\]%; \[77.45±2.15\]% vs \[66.87±0.73\]%,\[92.94±0.77\]% vs \[82.18±0.66\]%; \[51.78±0.63\]% vs \[43.54±095\]%; all P<0.05). Whereas, no significant change was observed in the granzyme B expression in the CD3+ cells \[(26.90±0.67）% vs (26.76±033）%，P>0.05\]. The cytolytic activity of CIK cells against EC9706 cells was increased significantly in the IL-12 group (E ∶T ratio of 20 ∶1, \[43.92± 1.67\]% vs \[35.34±1.22\]% ; E ∶T ratio of 30 ∶1, \[55.95±0.88\]% vs \[43.91±1.10\]%, all P<0.05). The cytotoxicity of CIK cells in the IL-12 and the control groups were significantly inhibited by anti-NKG2D monoclonal antibody (\[19.72±0.56\]% vs \[55.95±0.88\]%, \[19.83±1.20\]% vs \[43.91±1.10\]%, all P<0.05). Conclusion: IL-12 up-regulates the NKG2D and perforin expressions on CIK cells, enhancing their cytotoxicity against EC9706 cells.
2013, 20(2):201-206.
DOI: 10.3872/j.issn.1007-385X.2013.02.013
Abstract:
Objective: To construct a recombinant vector 3TAT-DRBD expressing purified fusion protein, and to preliminary validate its siRNA-binding activity and membrane-penetrating function. Methods: The target gene 3TAT-DRBD was obtained by gene synthesis and cloned to prokaryotic expression vector pET-44b. The expression of fusion protein was induced by IPTG. The fusion protein was purified by Ni-NTA agarose, and cut by thrombin and detected by Western blotting analysis. The binding activity of DRBD was tested by EMSA and the cytomembrane penetrating activity of TAT was observed by confocal laser scanning microscopy (CLSM). Results: Restriction enzyme digestion and gene sequencing showed that the recombinant plasmid pET-44b-3TAT-DRBD was successfully constructed. The fusion protein (containing Nus and S tags) induced by IPTG was efficiently expressed in E. coli, with the soluble parts accounting for around 80% of the total proteins. The tags were successfully cut off and the fusion protein without tags was purified with a molecular weight of 17 000 Da identified by Western blotting. EMSA identified that the fusion protein 3TAT-DRBD could effectively bind siRNA targeting survivin gene (survivin-siRNA). The efficiency of survivin-siRNA penetrating into prostate cancer PC3 cells mediated by TAT was significantly increased under an observation of CLSM. Conclusion: 3TAT-DRBD fusion protein with siRNA-binding activity and cell membrane-penetrating function is successfully expressed and purified, lying a good basis for further functional research and clinical application of 3TAT-DRBD.
Objective: To construct a recombinant vector 3TAT-DRBD expressing purified fusion protein, and to preliminary validate its siRNA-binding activity and membrane-penetrating function. Methods: The target gene 3TAT-DRBD was obtained by gene synthesis and cloned to prokaryotic expression vector pET-44b. The expression of fusion protein was induced by IPTG. The fusion protein was purified by Ni-NTA agarose, and cut by thrombin and detected by Western blotting analysis. The binding activity of DRBD was tested by EMSA and the cytomembrane penetrating activity of TAT was observed by confocal laser scanning microscopy (CLSM). Results: Restriction enzyme digestion and gene sequencing showed that the recombinant plasmid pET-44b-3TAT-DRBD was successfully constructed. The fusion protein (containing Nus and S tags) induced by IPTG was efficiently expressed in E. coli, with the soluble parts accounting for around 80% of the total proteins. The tags were successfully cut off and the fusion protein without tags was purified with a molecular weight of 17 000 Da identified by Western blotting. EMSA identified that the fusion protein 3TAT-DRBD could effectively bind siRNA targeting survivin gene (survivin-siRNA). The efficiency of survivin-siRNA penetrating into prostate cancer PC3 cells mediated by TAT was significantly increased under an observation of CLSM. Conclusion: 3TAT-DRBD fusion protein with siRNA-binding activity and cell membrane-penetrating function is successfully expressed and purified, lying a good basis for further functional research and clinical application of 3TAT-DRBD.
2013, 20(2):207-211.
DOI: 10.3872/j.issn.1007-385X.2013.02.014
Abstract:
Objective: Four siRNA fragments targeting kallikrein-related peptidase 7 ( KLK7 ) were synthesized in vitro. The most effective siRNA was selected, and the effect of silencing KLK7 expression on proliferation and apoptosis of gastric carcinoma AGS cells was observed. Methods: Four siRNA fragments targeting KLK7 (KLK7-siRNA-416, KLK7-siRNA-596, KLK7-siRNA-474 and KLK7-siRNA-795) were designed and transiently transfected into AGS cells. qRT-PCR was used to detect the expression of KLK7 mRNA in each interference group. Western blotting was used to detect the protein expression of HK7 (encoded by KLK7 gene). AGS cell proliferation, and the cells cycle and apoptosis after transfection were detected by MTT assay and flow cytometry, respectively. Results: Among four KLK7-siRNAs, KLK7-siRNA-416 showed the highest interference efficiency. The ratio of KLK7 mRNA expression in KLK7-siRNA-416 group was significantly lower than those in control group (\[0.32±0.049)% vs \[0.93±0.071\], P<0.01). The protein expression of HK7 in KLK7-siRNA-416 group after transfection for 48 h was significantly decreased (\[1.18±0.198\] vs \[0.52±0096\], P<0.01). The cell proliferation of KLK7-siRNA-416 group was significantly inhibited after transfection for 48 h, with inhibition rate of (37.70±0.12)%, P<0.05. Cell cycles were blocked in G0/G1 phase by transfection. However, no impact was found in AGS cell apoptosis. Conclusion: The silencing expression of KLK7 by KLK7-siRNA inhibited the AGS cell proliferation and block cell cycels in G0/G1 phase. However, no impact was found in AGS cell apoptosis.
Objective: Four siRNA fragments targeting kallikrein-related peptidase 7 ( KLK7 ) were synthesized in vitro. The most effective siRNA was selected, and the effect of silencing KLK7 expression on proliferation and apoptosis of gastric carcinoma AGS cells was observed. Methods: Four siRNA fragments targeting KLK7 (KLK7-siRNA-416, KLK7-siRNA-596, KLK7-siRNA-474 and KLK7-siRNA-795) were designed and transiently transfected into AGS cells. qRT-PCR was used to detect the expression of KLK7 mRNA in each interference group. Western blotting was used to detect the protein expression of HK7 (encoded by KLK7 gene). AGS cell proliferation, and the cells cycle and apoptosis after transfection were detected by MTT assay and flow cytometry, respectively. Results: Among four KLK7-siRNAs, KLK7-siRNA-416 showed the highest interference efficiency. The ratio of KLK7 mRNA expression in KLK7-siRNA-416 group was significantly lower than those in control group (\[0.32±0.049)% vs \[0.93±0.071\], P<0.01). The protein expression of HK7 in KLK7-siRNA-416 group after transfection for 48 h was significantly decreased (\[1.18±0.198\] vs \[0.52±0096\], P<0.01). The cell proliferation of KLK7-siRNA-416 group was significantly inhibited after transfection for 48 h, with inhibition rate of (37.70±0.12)%, P<0.05. Cell cycles were blocked in G0/G1 phase by transfection. However, no impact was found in AGS cell apoptosis. Conclusion: The silencing expression of KLK7 by KLK7-siRNA inhibited the AGS cell proliferation and block cell cycels in G0/G1 phase. However, no impact was found in AGS cell apoptosis.
2013, 20(2):212-216.
DOI: 10.3872/j.issn.1007-385X.2013.02.015
Abstract:
Objective:To explore the impact of Newcastle disease virus (lentogenic HBNU/LSRC/F3) strain on apoptosis of esophageal cancer ECA109 cells, and to compare with the other two strains (lentogenic LaSota strain and mesogenic Mukteswar strain) of their anti-tumor effects. Methods: ECA109 cells were cultured in vitro and infected by three NDV strains respectively. The proliferation inhibiting effects of NDVs on ECA109 cells were detected by CCK-8. The apoptosis of ECA109 cells after NDV infection was detected by laser confocal microscopy. Flow cytometry and agarose gel electrophoresis were used to detect the early and late apoptosis of ECA109 cells, respectively. Results: HBNU/LSRC/F3 NDV strain suppressed the proliferation of ECA109 cells, which was stronger than LaSota strain and weaker than Mukteswar strain (P<0.05). The proliferation inhibition rates of HBNU/LSRC/F3 strain, Mukteswar strain and LaSota strains on ECA109 cells were (39.87±1.99)%, (31.43±1.57)% and (19.89±0.99)%, respectively, infected by 5×10-4 Hu/ml NDV. Flow cytometry showed that the early apoptosis rate of ECA109 cells infected by HBNU/LSRC/F3 strain was (21.32±0.44)%, while Mukteswar strain was (22.27±0.23)% and LaSota strain was (14.32±0.61)%. Laser scanning confocal microscopy and DNA gel electrophoresis showed that Mukteswar strain primarily induced the late apoptosis. Whereas, HBNU/LSRC/F3 strain primarily induced the early apoptosis, which was more obvious than LaSota strain. Conclusion: Lentogenic HBNU/LSRC/F3 strain can effectively inhibit ECA109 cell proliferation and induce early apoptosis of ECA109 cells, which was lower than Mukteswar strain and significantly higher than LaSota strain. Therefore, HBNU/LSRC/F3 strain may have a high clinical value for anti-tumor therapy.
Objective:To explore the impact of Newcastle disease virus (lentogenic HBNU/LSRC/F3) strain on apoptosis of esophageal cancer ECA109 cells, and to compare with the other two strains (lentogenic LaSota strain and mesogenic Mukteswar strain) of their anti-tumor effects. Methods: ECA109 cells were cultured in vitro and infected by three NDV strains respectively. The proliferation inhibiting effects of NDVs on ECA109 cells were detected by CCK-8. The apoptosis of ECA109 cells after NDV infection was detected by laser confocal microscopy. Flow cytometry and agarose gel electrophoresis were used to detect the early and late apoptosis of ECA109 cells, respectively. Results: HBNU/LSRC/F3 NDV strain suppressed the proliferation of ECA109 cells, which was stronger than LaSota strain and weaker than Mukteswar strain (P<0.05). The proliferation inhibition rates of HBNU/LSRC/F3 strain, Mukteswar strain and LaSota strains on ECA109 cells were (39.87±1.99)%, (31.43±1.57)% and (19.89±0.99)%, respectively, infected by 5×10-4 Hu/ml NDV. Flow cytometry showed that the early apoptosis rate of ECA109 cells infected by HBNU/LSRC/F3 strain was (21.32±0.44)%, while Mukteswar strain was (22.27±0.23)% and LaSota strain was (14.32±0.61)%. Laser scanning confocal microscopy and DNA gel electrophoresis showed that Mukteswar strain primarily induced the late apoptosis. Whereas, HBNU/LSRC/F3 strain primarily induced the early apoptosis, which was more obvious than LaSota strain. Conclusion: Lentogenic HBNU/LSRC/F3 strain can effectively inhibit ECA109 cell proliferation and induce early apoptosis of ECA109 cells, which was lower than Mukteswar strain and significantly higher than LaSota strain. Therefore, HBNU/LSRC/F3 strain may have a high clinical value for anti-tumor therapy.
Yin Liangwei
,
Wang Suping
,
Zhang Li
,
Wang Ying
,
Ji Jun
,
Wang Heshuang
,
Guo Xu
,
Wang Xiaohong
,
Ma Shubei
,
Du Xiaohong
,
Ma Haiying
2013, 20(2):217-224.
DOI: 10.3872/j.issn.1007-385X.2013.02.016
Abstract:
Objective:To evaluate the therapeutic effect and safety of dendritic cells-cytokine induced killer cells (DC-CIK) combined with chemotherapy in the treatment of metastatic colorectal cancer (mCRC). Methods: 80 mCRC patients were selected from Dalian Centre Hospital during November 2010 to November 2011. 40 patients were treated by DC-CIK combined with chemotherapy (combined group), and another 40 patients were treated by chemotherapy alone (chemotherapy group). The immune function, therapeutic effect, toxicity and quality of life (QOL) were compared between the two groups after the treatment. Results: 160 cycles of DC-CIK were successfully treated. There were no obvious changes of T cell subsets in the peripheral blood in the combined group (P>0.05), while the ratios of CD3+, CD3+CD4+, CD3+ CD8+ and CD3-CD56+ cells were significantly decreased after the treatment in the chemotherapy group, and obviously lower than that in the combined group (P<0.05). After treatment for 3 cycles, the IFN-γ level of CD4+ T cells in the combined group was significantly increased (P<0.05), while the levels of IFN-γ, IL-2 and TNF-α in the chemotherapy group were significantly decreased after the treatment and were obviously lower than that in the combined group (P<005). No significant differences were found in the overall response rate (RR) between the combined group and the chemotherapy group (37.5% vs 22.5%,P>0.05). The disease control rate (DCR) of the combined group was significantly higher than that of the chemotherapy group (77.5% vs 50.0%, P<0.05). Ⅲ-Ⅳ grade leucopenia and tardily diarrhea in the combined group were obviously lower than those of the chemotherapy group (17.5% vs 42.5%, 50% vs 25.0%, P<0.05). Other side effects showed no significant differences between the two groups, which can be alleviated after symptomatic treatment (P>0.05). The median progression-free survival (PFS) of the combined group was longer than that of the chemotherapy group (6.5 months vs 4.5 months, P<0.05), while the median overall survival (OS) of the two groups had no significant difference (P>0.05). The physical and emotional functions of the combined group were better than those of the chemotherapy group (P<0.05). Conclusion: Treatment with DC-CIK adoptive immunotherapy combined with chemotherapy can effectively improve the immune function, improve the efficacy, reduce the side effects of chemotherapy, prolong the PFS, and improve QOL of mCRC patients.
Objective:To evaluate the therapeutic effect and safety of dendritic cells-cytokine induced killer cells (DC-CIK) combined with chemotherapy in the treatment of metastatic colorectal cancer (mCRC). Methods: 80 mCRC patients were selected from Dalian Centre Hospital during November 2010 to November 2011. 40 patients were treated by DC-CIK combined with chemotherapy (combined group), and another 40 patients were treated by chemotherapy alone (chemotherapy group). The immune function, therapeutic effect, toxicity and quality of life (QOL) were compared between the two groups after the treatment. Results: 160 cycles of DC-CIK were successfully treated. There were no obvious changes of T cell subsets in the peripheral blood in the combined group (P>0.05), while the ratios of CD3+, CD3+CD4+, CD3+ CD8+ and CD3-CD56+ cells were significantly decreased after the treatment in the chemotherapy group, and obviously lower than that in the combined group (P<0.05). After treatment for 3 cycles, the IFN-γ level of CD4+ T cells in the combined group was significantly increased (P<0.05), while the levels of IFN-γ, IL-2 and TNF-α in the chemotherapy group were significantly decreased after the treatment and were obviously lower than that in the combined group (P<005). No significant differences were found in the overall response rate (RR) between the combined group and the chemotherapy group (37.5% vs 22.5%,P>0.05). The disease control rate (DCR) of the combined group was significantly higher than that of the chemotherapy group (77.5% vs 50.0%, P<0.05). Ⅲ-Ⅳ grade leucopenia and tardily diarrhea in the combined group were obviously lower than those of the chemotherapy group (17.5% vs 42.5%, 50% vs 25.0%, P<0.05). Other side effects showed no significant differences between the two groups, which can be alleviated after symptomatic treatment (P>0.05). The median progression-free survival (PFS) of the combined group was longer than that of the chemotherapy group (6.5 months vs 4.5 months, P<0.05), while the median overall survival (OS) of the two groups had no significant difference (P>0.05). The physical and emotional functions of the combined group were better than those of the chemotherapy group (P<0.05). Conclusion: Treatment with DC-CIK adoptive immunotherapy combined with chemotherapy can effectively improve the immune function, improve the efficacy, reduce the side effects of chemotherapy, prolong the PFS, and improve QOL of mCRC patients.
17 Expression and clinical significance of fibroblast activation protein in non-small cell lung cancer
2013, 20(2):225-229.
DOI: 10.3872/j.issn.1007-385X.2013.02.017
Abstract:
Objective: To investigate the expression of fibroblast activation protein (FAP) mRNA in non-small cell lung cancer (NSCLC) tissue, and to evaluate its relationship with clinical characteristic and prognosis of NSCLC. Methods: Two hundred and forty-seven tumor specimens with NSCLC and forty-eight cases of corresponding adjacent tumor tissues selected from the Department of Biotherapy, Tumor Hospital Affiliated to Tianjing Medical University from June 2004 to December 2006. All cases were followed up until December 1, 2011. The FAP mRNA was detected by real-time PCR in NSCLC and normal tissues. Mann-Whitney-Wilcoxon U-Test was used to evaluate the association between clinicopathological parameters and FAP mRNA expression. Survival curves were plotted using Kaplan-Meier method, and survival difference was compared by the Log-rank, with Cox regression model to evaluate the independent prognostic factors. Results: FAP was over-expressed in NSCLC tissues. FAP mRNA expression level in NSCLC tissues were positively related to the Karnofsky Performance Score (KPS) (P<0.05), but not to the primary tumor location, size of tumor, lymph node metastasis, clinical stage and pathologic type of tumor (P>0.05). The median overall survival (OS) between patients with high FAP expression and low FAP expression showed no statistical significance (43 vs 39 months, P>0.05). Further histology hierarchical analysis showed that FAP mRNA expression in lung adenocarcinoma cancer patients was found to correlate inversely with clinical stages (P=0.031), and correlate positively with KPS (P=0.041). Lung adenocarcinoma lung cancer patients with high FAP expression had longer OS than did patients with low FAP expression (42 vs 26 months, P<0.05). Multivariate analyses showed that FAP expression was an independent prognostic predictor of lung adenocarcinoma cancer patients. Conclusion: FAP is over-expressed in NSCLC tissues. FAP is found to correlate inversely with clinical stage of lung adenocarcinoma cancer, and the expression of FAP is closely related to prognosis of lung adenocarcinoma cancer patients.
Objective: To investigate the expression of fibroblast activation protein (FAP) mRNA in non-small cell lung cancer (NSCLC) tissue, and to evaluate its relationship with clinical characteristic and prognosis of NSCLC. Methods: Two hundred and forty-seven tumor specimens with NSCLC and forty-eight cases of corresponding adjacent tumor tissues selected from the Department of Biotherapy, Tumor Hospital Affiliated to Tianjing Medical University from June 2004 to December 2006. All cases were followed up until December 1, 2011. The FAP mRNA was detected by real-time PCR in NSCLC and normal tissues. Mann-Whitney-Wilcoxon U-Test was used to evaluate the association between clinicopathological parameters and FAP mRNA expression. Survival curves were plotted using Kaplan-Meier method, and survival difference was compared by the Log-rank, with Cox regression model to evaluate the independent prognostic factors. Results: FAP was over-expressed in NSCLC tissues. FAP mRNA expression level in NSCLC tissues were positively related to the Karnofsky Performance Score (KPS) (P<0.05), but not to the primary tumor location, size of tumor, lymph node metastasis, clinical stage and pathologic type of tumor (P>0.05). The median overall survival (OS) between patients with high FAP expression and low FAP expression showed no statistical significance (43 vs 39 months, P>0.05). Further histology hierarchical analysis showed that FAP mRNA expression in lung adenocarcinoma cancer patients was found to correlate inversely with clinical stages (P=0.031), and correlate positively with KPS (P=0.041). Lung adenocarcinoma lung cancer patients with high FAP expression had longer OS than did patients with low FAP expression (42 vs 26 months, P<0.05). Multivariate analyses showed that FAP expression was an independent prognostic predictor of lung adenocarcinoma cancer patients. Conclusion: FAP is over-expressed in NSCLC tissues. FAP is found to correlate inversely with clinical stage of lung adenocarcinoma cancer, and the expression of FAP is closely related to prognosis of lung adenocarcinoma cancer patients.
2013, 20(2):230-236.
DOI: 10.3872/j.issn.1007-385X.2013.02.018
Abstract:
Objective:To isolate and identify the cancer stem cells from gastric cancer cell line SNU-5, and investigate the influence of CD90+ cancer stem cells on the metastasis and prognosis of gastric cancer. Methods: Non-adherent culture and PKH26 staining was used to determine whether there were cancer stem cells in human gastric cancer cell line SNU-5; Flow cytometry analysis was performed for identification of positive cells with stem cell markers within parent SNU-5 cells and sphere cells; CD90+ SNU-5 cells were sorted to identify its biological characteristics in vivo and tumorigenicity in vitro. Ninety-five cases with gastric cancer selected from cancer Hospital, immunohistochemistry was applied to detect CD90 expression in human gastric cancer. Results: The single PKH26 positive cell was observed in spheroids after SNU-5 cells were cultured for 11 days and CD90 could be co-stained with PKH26 in the spheroid cells, a dye indicating the characterization of stem cells. Serum sphere culture can enrich CD90+ SNU-5 cells by 6.1 times. The sorted CD90+ SNU-5 cells presented higher self-renewal ability (sphere formation rate: \[7.7±1.1\]% vs \[1.3±0.4\]%, \[18±03\]%，P<0.01), higher invasion (the number of invasive cells \[283.3±30.2\] vs \[48.0±7.5\], \[156.7±7.2\], P<0.01)than CD90- SUN-5 cells and parent SNU-5 cells. Only 2×102 CD90+ SNU-5 cells can form tumors at 6 weeks (4/6) in severe combined immune deficency (SCID) mice, whereas at least 2×104 CD90- SNU-5 cells were needed to form tumors at 10 weeks (1/6). Immunohistochemical results showed that CD90 expression was significantly associated with distant metastasis of gastric cancer (P<0.01). The survival rate of CD90 positive gastric cancer patients is obviously lower than that of CD90 negative patients (P<0.01). Conclusion: CD90+ cancer stem cells in huamn gastric cancer cell line SNU-5 show increased self-renewal and invasion abilitities, which are related to metastasis and prognosis of gastric cancer.
Objective:To isolate and identify the cancer stem cells from gastric cancer cell line SNU-5, and investigate the influence of CD90+ cancer stem cells on the metastasis and prognosis of gastric cancer. Methods: Non-adherent culture and PKH26 staining was used to determine whether there were cancer stem cells in human gastric cancer cell line SNU-5; Flow cytometry analysis was performed for identification of positive cells with stem cell markers within parent SNU-5 cells and sphere cells; CD90+ SNU-5 cells were sorted to identify its biological characteristics in vivo and tumorigenicity in vitro. Ninety-five cases with gastric cancer selected from cancer Hospital, immunohistochemistry was applied to detect CD90 expression in human gastric cancer. Results: The single PKH26 positive cell was observed in spheroids after SNU-5 cells were cultured for 11 days and CD90 could be co-stained with PKH26 in the spheroid cells, a dye indicating the characterization of stem cells. Serum sphere culture can enrich CD90+ SNU-5 cells by 6.1 times. The sorted CD90+ SNU-5 cells presented higher self-renewal ability (sphere formation rate: \[7.7±1.1\]% vs \[1.3±0.4\]%, \[18±03\]%，P<0.01), higher invasion (the number of invasive cells \[283.3±30.2\] vs \[48.0±7.5\], \[156.7±7.2\], P<0.01)than CD90- SUN-5 cells and parent SNU-5 cells. Only 2×102 CD90+ SNU-5 cells can form tumors at 6 weeks (4/6) in severe combined immune deficency (SCID) mice, whereas at least 2×104 CD90- SNU-5 cells were needed to form tumors at 10 weeks (1/6). Immunohistochemical results showed that CD90 expression was significantly associated with distant metastasis of gastric cancer (P<0.01). The survival rate of CD90 positive gastric cancer patients is obviously lower than that of CD90 negative patients (P<0.01). Conclusion: CD90+ cancer stem cells in huamn gastric cancer cell line SNU-5 show increased self-renewal and invasion abilitities, which are related to metastasis and prognosis of gastric cancer.
2013, 20(2):237-241.
DOI: 10.3872/j.issn.1007-385X.2013.02.019
Abstract:
Objective: To explore the clinical significance of the expressions of XIAP（X-linked inhibitor of apoptosis protein) and caspase-3 in colorectal adenocarcinoma and adenoma. Methods: Sixty-seven cases with colorectal adenocarcinoma, 30 cases of colorectal adenoma cases selected from the Department of Pathology, First Affiliated Hospital of Liaoning Medical College from 2010 to 2012 with surgical resection, and 30 cases of corresponding adjacent mucosa (the distance from the edge of the cancerous tissue 5 cm) were used as a control. Immunohistochemistry was used to detect the expressions of XIAP and caspase-3 proteins in colorectal adenocarcinoma and adenoma tissues; Western blotting was used to detect the expression of XIAP in the colorectal adenocarcinoma and adenoma tissues; The relationship between the expression of XIAP and the clinical pathology parameters of colorectal adenocarcinoma was analyzed. Results: The positive rate of XIAP expression in the colorectal adenoma group (71.6%) was higher than that in colorectal adenocarcinoma (46.7%), and its expression rate was increasing with the decrease of the tissue differentiation degree (χ2=16.132, P<0.05); the positive rate of caspase-3 expression in the colorectal adenocarcinoma tissues (18.0%) was lower than that in the colorectal adenoma group (43.3%), and its expression rate was decreased with the decrease of the pathological differentiation degree (P<0.05). The expression of XIAP protein was in a negative correlation with that of caspase-3 (r=-0.396, P<0.05). Conclusion: The XIAP protein might play a significant role in promoting the progress from colorectal adenoma to colorectal adenocarcinoma by inhibiting caspase-3.
Objective: To explore the clinical significance of the expressions of XIAP（X-linked inhibitor of apoptosis protein) and caspase-3 in colorectal adenocarcinoma and adenoma. Methods: Sixty-seven cases with colorectal adenocarcinoma, 30 cases of colorectal adenoma cases selected from the Department of Pathology, First Affiliated Hospital of Liaoning Medical College from 2010 to 2012 with surgical resection, and 30 cases of corresponding adjacent mucosa (the distance from the edge of the cancerous tissue 5 cm) were used as a control. Immunohistochemistry was used to detect the expressions of XIAP and caspase-3 proteins in colorectal adenocarcinoma and adenoma tissues; Western blotting was used to detect the expression of XIAP in the colorectal adenocarcinoma and adenoma tissues; The relationship between the expression of XIAP and the clinical pathology parameters of colorectal adenocarcinoma was analyzed. Results: The positive rate of XIAP expression in the colorectal adenoma group (71.6%) was higher than that in colorectal adenocarcinoma (46.7%), and its expression rate was increasing with the decrease of the tissue differentiation degree (χ2=16.132, P<0.05); the positive rate of caspase-3 expression in the colorectal adenocarcinoma tissues (18.0%) was lower than that in the colorectal adenoma group (43.3%), and its expression rate was decreased with the decrease of the pathological differentiation degree (P<0.05). The expression of XIAP protein was in a negative correlation with that of caspase-3 (r=-0.396, P<0.05). Conclusion: The XIAP protein might play a significant role in promoting the progress from colorectal adenoma to colorectal adenocarcinoma by inhibiting caspase-3.
2013, 20(2):242-246.
DOI: 10.3872/j.issn.1007-385X.2013.02.020
Abstract:
黑素瘤相关抗原（melanoma-associated antigen，MAGE）-A基因亚家族属于癌睾丸抗原家族，其定位于X染色体，包括MAGE-A1～MAGE-A12共12个成员。对MAGE-A在基因和蛋白水平的研究发现，其在正常组织中几乎不表达，而在多种肿瘤组织中均有较高水平的表达，并且往往表达一种以上的MAGE-A亚型。正常情况下，由于CpG岛高度甲基化，MAGE-A基因沉默表达。受辐射等因素影响时，基因组易发生去甲基化，促使MAGE-A基因表达。组蛋白的乙酰化也与MAGE-A的表达有关。MAGE-A作为肿瘤相关抗原，与肿瘤的发生、发展、耐药及较差的预后密切相关。由于MAGE-A特异性的表达于多种肿瘤，对MAGE-A亚型的检测尤其是多种亚型的联合检测有助于MAGE-A阳性肿瘤的诊断。此外，MAGE-A基因编码的抗原肽由MHCⅠ分子提呈至细胞毒性T细胞，从而发挥抗肿瘤活性。应用MAGE-A抗原肽的肿瘤疫苗已经投入临床试验，并取得了良好的治疗效果。
黑素瘤相关抗原（melanoma-associated antigen，MAGE）-A基因亚家族属于癌睾丸抗原家族，其定位于X染色体，包括MAGE-A1～MAGE-A12共12个成员。对MAGE-A在基因和蛋白水平的研究发现，其在正常组织中几乎不表达，而在多种肿瘤组织中均有较高水平的表达，并且往往表达一种以上的MAGE-A亚型。正常情况下，由于CpG岛高度甲基化，MAGE-A基因沉默表达。受辐射等因素影响时，基因组易发生去甲基化，促使MAGE-A基因表达。组蛋白的乙酰化也与MAGE-A的表达有关。MAGE-A作为肿瘤相关抗原，与肿瘤的发生、发展、耐药及较差的预后密切相关。由于MAGE-A特异性的表达于多种肿瘤，对MAGE-A亚型的检测尤其是多种亚型的联合检测有助于MAGE-A阳性肿瘤的诊断。此外，MAGE-A基因编码的抗原肽由MHCⅠ分子提呈至细胞毒性T细胞，从而发挥抗肿瘤活性。应用MAGE-A抗原肽的肿瘤疫苗已经投入临床试验，并取得了良好的治疗效果。
2013, 20(2):247-250.
DOI: 10.3872/j.issn.1007-385X.2013.02.021
Abstract:
结直肠癌是人类最常见的恶性肿瘤之一，分子靶向治疗药物近年来在对晚期结直肠癌患者的综合治疗中发挥了重要作用，因此结直肠癌的靶向治疗已经成为临床研究的热点。目前晚期结直肠癌靶向治疗的代表药物主要有两大类，一类是以血管内皮生长因子为作用靶点的贝伐单抗(bevacizumab)等药物，通过抑制肿瘤血管的新生来抑制肿瘤的生长；另一类是以表皮生长因子受体为作用靶点的西妥昔单抗(cetuximab)及帕尼单抗(panitumumab)等药物，通过与肿瘤细胞表面的表皮生长因子受体特异性结合来竞争性阻断其与其他配体的结合来达到抑制肿瘤生长的目的。两类分子靶向治疗药物的作用靶点各不相同，因此在临床治疗过程中所产生的疗效也不同，且分子靶向治疗药物在临床上的应用尚存在许多问题有待进一步阐明。
结直肠癌是人类最常见的恶性肿瘤之一，分子靶向治疗药物近年来在对晚期结直肠癌患者的综合治疗中发挥了重要作用，因此结直肠癌的靶向治疗已经成为临床研究的热点。目前晚期结直肠癌靶向治疗的代表药物主要有两大类，一类是以血管内皮生长因子为作用靶点的贝伐单抗(bevacizumab)等药物，通过抑制肿瘤血管的新生来抑制肿瘤的生长；另一类是以表皮生长因子受体为作用靶点的西妥昔单抗(cetuximab)及帕尼单抗(panitumumab)等药物，通过与肿瘤细胞表面的表皮生长因子受体特异性结合来竞争性阻断其与其他配体的结合来达到抑制肿瘤生长的目的。两类分子靶向治疗药物的作用靶点各不相同，因此在临床治疗过程中所产生的疗效也不同，且分子靶向治疗药物在临床上的应用尚存在许多问题有待进一步阐明。
2013, 20(2):251-254.
DOI: 10.3872/j.issn.1007-385X.2013.02.022
Abstract:
紫杉醇（paclitaxel, PTX）是一种有丝分裂抑制剂，通过促进微管蛋白聚合、抑制解聚、保持微管蛋白稳定、抑制细胞有丝分裂和促进肿瘤细胞凋亡发挥抗肿瘤作用。通过研究紫杉醇的结构和功能，发现紫杉醇对免疫细胞包括效应性T细胞、树突状细胞（dendritic cell, DC）、自然杀伤细胞（Natural cell, NK）、调节性T细胞（regulatory T cell, Treg）和巨噬细胞（macrophage）等具有广泛调节作用，紫杉醇杀伤肿瘤细胞的同时逆转肿瘤的免疫逃逸。紫杉醇与过继细胞免疫治疗联合可减轻化疗的不良反应、增加化疗效果。紫杉醇在非小细胞肺癌化疗中抑制调节性T细胞的数量和功能，在乳腺癌化疗过程中增强NK细胞抗肿瘤活性，在卵巢癌化疗中增加肿瘤抗原的免疫原性及促进DC的抗原提呈作用。总之，紫杉醇的免疫调节功能在抗肿瘤领域有广泛的应用前景。
紫杉醇（paclitaxel, PTX）是一种有丝分裂抑制剂，通过促进微管蛋白聚合、抑制解聚、保持微管蛋白稳定、抑制细胞有丝分裂和促进肿瘤细胞凋亡发挥抗肿瘤作用。通过研究紫杉醇的结构和功能，发现紫杉醇对免疫细胞包括效应性T细胞、树突状细胞（dendritic cell, DC）、自然杀伤细胞（Natural cell, NK）、调节性T细胞（regulatory T cell, Treg）和巨噬细胞（macrophage）等具有广泛调节作用，紫杉醇杀伤肿瘤细胞的同时逆转肿瘤的免疫逃逸。紫杉醇与过继细胞免疫治疗联合可减轻化疗的不良反应、增加化疗效果。紫杉醇在非小细胞肺癌化疗中抑制调节性T细胞的数量和功能，在乳腺癌化疗过程中增强NK细胞抗肿瘤活性，在卵巢癌化疗中增加肿瘤抗原的免疫原性及促进DC的抗原提呈作用。总之，紫杉醇的免疫调节功能在抗肿瘤领域有广泛的应用前景。
2013, 20(2):255-258.
DOI: 10.3872/j.issn.1007-385X.2013.02.023
Abstract:
MicroRNA(miRNA)是一种内源性非编码小RNA，在转录后水平负调节靶基因的表达，并参与多种细胞的生命活动，特别是在肿瘤的发生、发展和迁移过程中起重要作用。miRNA通过调控相关靶基因，影响黑素瘤相关蛋白的转录翻译，从而促进或抑制黑素瘤的发生、发展和转移。miR-137和miR-182可以负向调节小眼球相关转录因子（microphthalmia-associated transcription factor,MITF)表达，抑制黑素瘤增殖和侵袭；MITF也可以反向调控miR-137和miR-182，增强黑素瘤的迁移能力。miR-221和miR-222通过下调p27Kipl/CDKN1B和c-kit受体两条信号通路促进黑素细胞恶化，从而在黑素瘤中发挥癌基因样效应；而在早幼粒白血病锌指蛋白（promyelocytic leukemia Zinc-Finger, PLZF)结合miR-221和miR-222的调节区后，可以抑制黑素瘤细胞转化和侵袭。Let-7家族成员主要抑制黑素瘤发生、发展和迁移，一旦缺失或表达下降则促进黑素瘤增殖发展。miR-34通过下调原癌基因和相关细胞周期蛋白表达，在黑素瘤中主要发挥抑癌基因样作用。通过检测正常个体和黑素瘤患者外周血miRNA表达谱的差异，有助于诊断黑素瘤；监测分析相关miRNA变化有助于判断患者的疗效和预后情况。因此，深入探索相关miRNA在黑素瘤中的靶基因及调控作用机制有望为黑素瘤的诊断、治疗开辟新途径。
MicroRNA(miRNA)是一种内源性非编码小RNA，在转录后水平负调节靶基因的表达，并参与多种细胞的生命活动，特别是在肿瘤的发生、发展和迁移过程中起重要作用。miRNA通过调控相关靶基因，影响黑素瘤相关蛋白的转录翻译，从而促进或抑制黑素瘤的发生、发展和转移。miR-137和miR-182可以负向调节小眼球相关转录因子（microphthalmia-associated transcription factor,MITF)表达，抑制黑素瘤增殖和侵袭；MITF也可以反向调控miR-137和miR-182，增强黑素瘤的迁移能力。miR-221和miR-222通过下调p27Kipl/CDKN1B和c-kit受体两条信号通路促进黑素细胞恶化，从而在黑素瘤中发挥癌基因样效应；而在早幼粒白血病锌指蛋白（promyelocytic leukemia Zinc-Finger, PLZF)结合miR-221和miR-222的调节区后，可以抑制黑素瘤细胞转化和侵袭。Let-7家族成员主要抑制黑素瘤发生、发展和迁移，一旦缺失或表达下降则促进黑素瘤增殖发展。miR-34通过下调原癌基因和相关细胞周期蛋白表达，在黑素瘤中主要发挥抑癌基因样作用。通过检测正常个体和黑素瘤患者外周血miRNA表达谱的差异，有助于诊断黑素瘤；监测分析相关miRNA变化有助于判断患者的疗效和预后情况。因此，深入探索相关miRNA在黑素瘤中的靶基因及调控作用机制有望为黑素瘤的诊断、治疗开辟新途径。
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- 《中国肿瘤生物治疗杂志》
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- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
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- 刊号：ISSN 1007-385X
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.