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Volume 17,Issue 2,2010 Table of Contents
2010, 17(2):115-120.
DOI: 10.3872/j.issn.1007-385X.2010.2.001
Abstract:
Abstract Immunotherapy as a promising therapy strategy is expected to bring new hope for breast cancer treatment. Recently several breast cancer vaccines have been developed. Rational and rapid application of these scientific discoveries in clinical therapies for breast cancer patients poses great challenges and opportunities. This article is aimed to elaborate on the design strategies, current clinical trials, and future progression and application of breast cancer therapeutic vaccines. With the deepening of our understanding on the tumor-specific immune response and tumor microenvironment, and continuous discovery of new tumor-specific antigens, the research level of breast cancer vaccines will be further improved and will benefit the clinical treatment of breast cancer.
Abstract Immunotherapy as a promising therapy strategy is expected to bring new hope for breast cancer treatment. Recently several breast cancer vaccines have been developed. Rational and rapid application of these scientific discoveries in clinical therapies for breast cancer patients poses great challenges and opportunities. This article is aimed to elaborate on the design strategies, current clinical trials, and future progression and application of breast cancer therapeutic vaccines. With the deepening of our understanding on the tumor-specific immune response and tumor microenvironment, and continuous discovery of new tumor-specific antigens, the research level of breast cancer vaccines will be further improved and will benefit the clinical treatment of breast cancer.
SUN Li-chao
,
ZHAO Xuan
,
YU Long
,
HAN Lu-lu
,
LIU Tong
,
HU Hai
,
SUN Li-xin
,
YANG Zhi-hua
,
RAN Yu-liang
2010, 17(2):121-127.
DOI: 10.3872/j.issn.1007-385X.2010.2.002
Abstract:
Abstract Objective:To prepare functional monoclonal antibodies(McAbs) against liver cancer stem cells, so as to provide candidate antibody drugs for stem cell-targeted therapy of liver cancer. Methods: Human liver cancer stem-like cells (hLCSLCs) were separated from human hepatocarcinoma tissues and were used to immunize BALB/c nude mice. Spleen cells from hLCSLCs-immunized mice were fused with SP2/0 cells to prepare large monoclonal antibody library. Hybridoma McAbs recognizing hLCSLCs were screened and identified by immunofluorescence, sphere formation culture and in vivo tumor formation assays. hLCSLCs side population cells (hLCSLCs-SP) were sorted by flow cytometry. The effects of hybridoma McAbs on self-renewal and proliferation of hLCSLCs-SP were identified by serum-free suspension culture and CCK-8 assay. Results: A total of 2 964 McAb clones were obtained by fusing immunized spleen cells with SP2/0 cells, and 237 McAbs could interact with hLCSLCs as detected by fixed-cell immunofluorescence; 116 of the 237 McAbs interacted with the membrane of hLCSLCs, and 33 McAbs specifically reacted with hLCSLCs-SP but not with non-hLCSLCs-SP, with positive rates being 2%-5%. Six of the 33 McAbs co-stained with CD133 on hLCSLCs-SP. Further investigation showed that the positive rates of these 6 McAbs were 3%-26% with sphere cells after serum-free suspension culture, which were significantly higher than those with hLCSLCs-SP. Tumor formation rate was 100% when 1×104 hybridoma clone 15D2-positive hLCSLCs were injected into nude mice. Functional study showed that 4 of these 6 McAbs significantly suppressed the proliferation and sphere formation ability of hLCSLCs-SP, with the inhibitory rates being 24%-42% and 13%-50%, respectively. Conclusion: We have successfully constructed the large McAb library against hLCSLCs, from which 4 hybridoma McAbs can specifically react with hLCSLCs-SP, laying a foundation for cancer stem cell-based antibody-targeted therapy for liver cancer.
Abstract Objective:To prepare functional monoclonal antibodies(McAbs) against liver cancer stem cells, so as to provide candidate antibody drugs for stem cell-targeted therapy of liver cancer. Methods: Human liver cancer stem-like cells (hLCSLCs) were separated from human hepatocarcinoma tissues and were used to immunize BALB/c nude mice. Spleen cells from hLCSLCs-immunized mice were fused with SP2/0 cells to prepare large monoclonal antibody library. Hybridoma McAbs recognizing hLCSLCs were screened and identified by immunofluorescence, sphere formation culture and in vivo tumor formation assays. hLCSLCs side population cells (hLCSLCs-SP) were sorted by flow cytometry. The effects of hybridoma McAbs on self-renewal and proliferation of hLCSLCs-SP were identified by serum-free suspension culture and CCK-8 assay. Results: A total of 2 964 McAb clones were obtained by fusing immunized spleen cells with SP2/0 cells, and 237 McAbs could interact with hLCSLCs as detected by fixed-cell immunofluorescence; 116 of the 237 McAbs interacted with the membrane of hLCSLCs, and 33 McAbs specifically reacted with hLCSLCs-SP but not with non-hLCSLCs-SP, with positive rates being 2%-5%. Six of the 33 McAbs co-stained with CD133 on hLCSLCs-SP. Further investigation showed that the positive rates of these 6 McAbs were 3%-26% with sphere cells after serum-free suspension culture, which were significantly higher than those with hLCSLCs-SP. Tumor formation rate was 100% when 1×104 hybridoma clone 15D2-positive hLCSLCs were injected into nude mice. Functional study showed that 4 of these 6 McAbs significantly suppressed the proliferation and sphere formation ability of hLCSLCs-SP, with the inhibitory rates being 24%-42% and 13%-50%, respectively. Conclusion: We have successfully constructed the large McAb library against hLCSLCs, from which 4 hybridoma McAbs can specifically react with hLCSLCs-SP, laying a foundation for cancer stem cell-based antibody-targeted therapy for liver cancer.
CHONG Jing-hui
,
TIAN Chen
,
SHI Ying-xu
,
WANG Jin-hong
,
LIN Yong-min
,
XU Jing
,
WU Ke-fu
,
ZHENG Guo-guang
2010, 17(2):128-133.
DOI: 10.3872/j.issn.1007-385X.2010.2.003
Abstract:
Abstract Objective:To investigate the expression features of P2X family receptors during the development and progression of murine acute T lymphoblastic leukemia. Methods: A Notch1 over-expressing murine T cell induced-acute lymphoblastic leukemia model was prepared. CD45.2+GFP+ leukemia cells were sorted by flow cytometry. The expressions of P2X family receptors were determined by real-time PCR. P2X7 receptor-mediated intracellular Ca 2+ change was measured by fluorescent spectrophotometer. Results: Mouse bone marrow cells (BMNCs) of both control and leukemia mice expressed all P2X receptors except for P2X5. P2X7 receptor expression increased gradually during the induction of T lymphoblastic leukemia; P2X1 and P2X3 receptors decreased; and P2X2, P2X4 and P2X6 receptors had no detectable changes. Similar expression patterns were observed in sorted CD45.2+GFP+ leukemia cells. P2X7 receptor-mediated calcium response was detected in BMNCs from both leukemia and control mice; and the response could be blocked by its specific antagonist KN62. However, the calcium response showed different patterns: it sustained an increase in leukemia group, whereas gradually decreased after reaching peak in the control group. Conclusion: P2X1, P2X3 and P2X7 receptor expressions are related to the development and progression of murine acute T lymphoblastic leukemia, suggesting that intercellular communications mediated by these receptors may play important roles in leukemia.
Abstract Objective:To investigate the expression features of P2X family receptors during the development and progression of murine acute T lymphoblastic leukemia. Methods: A Notch1 over-expressing murine T cell induced-acute lymphoblastic leukemia model was prepared. CD45.2+GFP+ leukemia cells were sorted by flow cytometry. The expressions of P2X family receptors were determined by real-time PCR. P2X7 receptor-mediated intracellular Ca 2+ change was measured by fluorescent spectrophotometer. Results: Mouse bone marrow cells (BMNCs) of both control and leukemia mice expressed all P2X receptors except for P2X5. P2X7 receptor expression increased gradually during the induction of T lymphoblastic leukemia; P2X1 and P2X3 receptors decreased; and P2X2, P2X4 and P2X6 receptors had no detectable changes. Similar expression patterns were observed in sorted CD45.2+GFP+ leukemia cells. P2X7 receptor-mediated calcium response was detected in BMNCs from both leukemia and control mice; and the response could be blocked by its specific antagonist KN62. However, the calcium response showed different patterns: it sustained an increase in leukemia group, whereas gradually decreased after reaching peak in the control group. Conclusion: P2X1, P2X3 and P2X7 receptor expressions are related to the development and progression of murine acute T lymphoblastic leukemia, suggesting that intercellular communications mediated by these receptors may play important roles in leukemia.
2010, 17(2):134-138.
DOI: 10.3872/j.issn.1007-385X.2010.2.004
Abstract:
Abstract Objective:To investigate the apoptosis induction effect of oridonin (Ori) on human multiple myeloma cell line ARH-77 and the possible mechanism. Methods: ARH-77 cells were treated with different concentrations of oridonin (2.5, 5 and 10 μmol/L), and then the proliferation of ARH-77 cells was detected by MTT assay. The morphological change of apoptotic ARH-77 cells was observed under phase contrast microscope and Hoechst 33258 staining. The apoptosis and the change of mitochondrial membrane potential (Δψm) of ARH-77 cells were examined by flow cytometry. The caspase 8 activity was measured by caspase-8 apoptosis kit. Results: Ori significantly inhibited the growth of ARH-77 cells in a time- and dose-dependent manner. After treatment with 10 μmol/L Ori for 24 h, ARH-77 cells became shrunk, with cytoplasm vacuoles and apoptotic bodies. Flow cytometry results revealed that the apoptotic rates of ARH-77 cells after Ori treatments (2.5, 5 and 10 μmol/L) were significantly higher than that of control group, with apoptotic rates being (15.07±0.78)%, (21.00±1.49)%, (27.45±2.47)% vs (5.27±1.46)% (P<0.01). Green fluorescence percentages of Δψm of ARH-77 cells were significantly increased in Ori-treated groups (2.5, 5 and 10 μmol/L) compared with that in control group, with percentages being (26.80±0.75)%, (36.81±2.27)%, (49.48±1.10)% vs (16.96±0.50)% (P<0.01). Meanwhile, caspase 8 activity of ARH-77 cells was significantly up-regulated by Ori treatment (P< 0.01). Conclusion: Oridonin can markedly induce apoptosis of human multiple myeloma ARH-77 cells, which might be related to extrinsic and intrinsic pathways.
Abstract Objective:To investigate the apoptosis induction effect of oridonin (Ori) on human multiple myeloma cell line ARH-77 and the possible mechanism. Methods: ARH-77 cells were treated with different concentrations of oridonin (2.5, 5 and 10 μmol/L), and then the proliferation of ARH-77 cells was detected by MTT assay. The morphological change of apoptotic ARH-77 cells was observed under phase contrast microscope and Hoechst 33258 staining. The apoptosis and the change of mitochondrial membrane potential (Δψm) of ARH-77 cells were examined by flow cytometry. The caspase 8 activity was measured by caspase-8 apoptosis kit. Results: Ori significantly inhibited the growth of ARH-77 cells in a time- and dose-dependent manner. After treatment with 10 μmol/L Ori for 24 h, ARH-77 cells became shrunk, with cytoplasm vacuoles and apoptotic bodies. Flow cytometry results revealed that the apoptotic rates of ARH-77 cells after Ori treatments (2.5, 5 and 10 μmol/L) were significantly higher than that of control group, with apoptotic rates being (15.07±0.78)%, (21.00±1.49)%, (27.45±2.47)% vs (5.27±1.46)% (P<0.01). Green fluorescence percentages of Δψm of ARH-77 cells were significantly increased in Ori-treated groups (2.5, 5 and 10 μmol/L) compared with that in control group, with percentages being (26.80±0.75)%, (36.81±2.27)%, (49.48±1.10)% vs (16.96±0.50)% (P<0.01). Meanwhile, caspase 8 activity of ARH-77 cells was significantly up-regulated by Ori treatment (P< 0.01). Conclusion: Oridonin can markedly induce apoptosis of human multiple myeloma ARH-77 cells, which might be related to extrinsic and intrinsic pathways.
MA Jun-fen
,
LIU Kang-dong
,
LIU Xia
,
YANG Hong-yan
,
HUANG You-tian
,
ZHAO Ming-yao
,
DONG Zi-ming
2010, 17(2):139-143.
DOI: 10.3872/j.issn.1007-385X.2010.2.005
Abstract:
Abstract Objective:To observe the effect of trichostatin A (TSA) on Coxsachievirus and adenovirus receptor (CAR) expression in membrane of human esophageal cancer EC1 cells, and to discuss the role of MAPK/ERK signal pathway in the up-regulation of CAR expression triggered by TSA. Methods: EC1 cells were treated with TSA (0.3, 0.5, 1.0 μmol/L), and CAR expressions on EC1 cells were examined by immunofluorescence staining, RT-PCR and Western blotting analysis. EC1 cells were also treated with 1.0 μmol/L TSA for 0, 1, 6, 12, 24, and 48 h, and then the CAR expression and phosphorylation of ERK were detected by Western blotting analysis. The correlation between ERK phosphorylation level and the CAR expression was analyzed. Results: CAR protein and mRNA expressions in EC1 cells were significantly increased after treatment with 0.3, 0.5, and 1.0 μmol/L TSA (P<0.05), and the increase was in a dose-dependent manner. EC1 cells treated with 1.0 μmol/L TSA for different time periods also showed significantly increased CAR expression (P<0.05), while p-ERK expression levels in EC1 cells were remarkably decreased. The expression of p-ERK in EC1 cells treated with TSA was negatively correlated with that of CAR (r=-0.886, P<0.01). Conclusion: TSA can increase the expression of CAR in human EC1 cells, and the possible mechanisms may be related to the inhibition of ERK/MAPK pathway in EC1 cells.
Abstract Objective:To observe the effect of trichostatin A (TSA) on Coxsachievirus and adenovirus receptor (CAR) expression in membrane of human esophageal cancer EC1 cells, and to discuss the role of MAPK/ERK signal pathway in the up-regulation of CAR expression triggered by TSA. Methods: EC1 cells were treated with TSA (0.3, 0.5, 1.0 μmol/L), and CAR expressions on EC1 cells were examined by immunofluorescence staining, RT-PCR and Western blotting analysis. EC1 cells were also treated with 1.0 μmol/L TSA for 0, 1, 6, 12, 24, and 48 h, and then the CAR expression and phosphorylation of ERK were detected by Western blotting analysis. The correlation between ERK phosphorylation level and the CAR expression was analyzed. Results: CAR protein and mRNA expressions in EC1 cells were significantly increased after treatment with 0.3, 0.5, and 1.0 μmol/L TSA (P<0.05), and the increase was in a dose-dependent manner. EC1 cells treated with 1.0 μmol/L TSA for different time periods also showed significantly increased CAR expression (P<0.05), while p-ERK expression levels in EC1 cells were remarkably decreased. The expression of p-ERK in EC1 cells treated with TSA was negatively correlated with that of CAR (r=-0.886, P<0.01). Conclusion: TSA can increase the expression of CAR in human EC1 cells, and the possible mechanisms may be related to the inhibition of ERK/MAPK pathway in EC1 cells.
2010, 17(2):144-148.
DOI: 10.3872/j.issn.1007-385X.2010.2.006
Abstract:
Abstract Objective:To investigate the relationship between epithelial-mesenchymal transition (EMT) and P-glucoprotein (P-gp)-induced multidrug resistance (MDR) in breast cancer cells and the corresponding mechanisms. Methods: Eukaryotic expression vector pcDNA-Snail was constructed and then transfected into human breast cancer cell line MCF-7. Multidrug resistance was induced by doxorubicin (DOX) in different groups. Expressions of epithelial marker E-cadherin, interstitial marker vimentin, and P-glucoprotein (P-gp) were detected by immunofluorescence. MTT assay was used to measure the proliferation of drug resistant MCF-7 cells. Expressions of Snail, MDR1, and p38-MAPK mRNA were evaluated by RT-PCR. Results: Immunofluorescence showed that MCF-7 cells had EMT after transfection with pcDNA-Snail vector. The expression of E-cadherin was downregulated, and expressions of vimentin and P-gp were upregulated in EMT-like MCF-7 cells. Drug resistance of MCF-7/Snail cells was significantly enhanced compared with MCF-7 cells after induction by DOX (P<0.05). The expressions of MDR1 and p38-MAPK mRNA in EMT-like cells were also significantly increased compared with those in parental MCF-7 cells (P<0.05). Conclusion: EMT may trigger DOX-induced and P-gp-mediated MDR via p38-MAPK in MCF-7 cells.
Abstract Objective:To investigate the relationship between epithelial-mesenchymal transition (EMT) and P-glucoprotein (P-gp)-induced multidrug resistance (MDR) in breast cancer cells and the corresponding mechanisms. Methods: Eukaryotic expression vector pcDNA-Snail was constructed and then transfected into human breast cancer cell line MCF-7. Multidrug resistance was induced by doxorubicin (DOX) in different groups. Expressions of epithelial marker E-cadherin, interstitial marker vimentin, and P-glucoprotein (P-gp) were detected by immunofluorescence. MTT assay was used to measure the proliferation of drug resistant MCF-7 cells. Expressions of Snail, MDR1, and p38-MAPK mRNA were evaluated by RT-PCR. Results: Immunofluorescence showed that MCF-7 cells had EMT after transfection with pcDNA-Snail vector. The expression of E-cadherin was downregulated, and expressions of vimentin and P-gp were upregulated in EMT-like MCF-7 cells. Drug resistance of MCF-7/Snail cells was significantly enhanced compared with MCF-7 cells after induction by DOX (P<0.05). The expressions of MDR1 and p38-MAPK mRNA in EMT-like cells were also significantly increased compared with those in parental MCF-7 cells (P<0.05). Conclusion: EMT may trigger DOX-induced and P-gp-mediated MDR via p38-MAPK in MCF-7 cells.
2010, 17(2):149-154.
DOI: 10.3872/j.issn.1007-385X.2010.2.007
Abstract:
Abstract Objective:To isolate, culture and identify the colon cancer stem-like cells with CD44+/EPCAM high phenotype from LoVo cell line, and to observe their biological behaviors and verify the existence of tumor stem cells in LoVo cell line. Methods: CD44+/EPCAM high cells were sorted from LoVo cells cultured in common-serum medium by flow cytometry, and the resultants were further cultured in serum-free medium (SFM) supplemented with growth factors. The proliferation and differentiation of CD44+/EPCAM high cells were observed. The proliferations and cell cycle distributions of CD44+/EPCAM high, EPCAM low, and un-sorted LoVo cells were estimated by MTT and flow cytometry, respectively. Nude mice were implanted with the above 3 different cells, and tumor formation rates of different groups were analyzed. Expressions of CD44 and EPCAM in the second passage of CD44+/EPCAM high cells were determined by immunofluorescence staining. Results: We found that 17.4% of LoVo cells were CD44+/EPCAM high cells, which could steadily proliferate and assemble into tumor cell spheres in SFM supplemented with growth factors. CD44+/EPCAM high cells could differentiate into adherent cells by serum, similar to LoVo cells. The proliferation capacity of CD44+/EPCAM high cells was higher than those of EPCAM low and LoVo cells, and the cell cycle of CD44+/EPCAM high cells was mostly in G0/G1 phase. Tumor formation rate of 500 CD44+/EPCAM high cells was 90% (9/10) in nude mice, with 1×104 EPCAM low cells being 0% (0/10). Moreover, expressions of CD44 and EPCAM in the second passage of CD44+/EPCAM high cells could still be detected. Conclusion: Colon cancer cell line LoVo contains CD44+/EPCAM high stem-like cells, and CD44+/EPCAM high cells can be used in further research of colon cancer stem cells.
Abstract Objective:To isolate, culture and identify the colon cancer stem-like cells with CD44+/EPCAM high phenotype from LoVo cell line, and to observe their biological behaviors and verify the existence of tumor stem cells in LoVo cell line. Methods: CD44+/EPCAM high cells were sorted from LoVo cells cultured in common-serum medium by flow cytometry, and the resultants were further cultured in serum-free medium (SFM) supplemented with growth factors. The proliferation and differentiation of CD44+/EPCAM high cells were observed. The proliferations and cell cycle distributions of CD44+/EPCAM high, EPCAM low, and un-sorted LoVo cells were estimated by MTT and flow cytometry, respectively. Nude mice were implanted with the above 3 different cells, and tumor formation rates of different groups were analyzed. Expressions of CD44 and EPCAM in the second passage of CD44+/EPCAM high cells were determined by immunofluorescence staining. Results: We found that 17.4% of LoVo cells were CD44+/EPCAM high cells, which could steadily proliferate and assemble into tumor cell spheres in SFM supplemented with growth factors. CD44+/EPCAM high cells could differentiate into adherent cells by serum, similar to LoVo cells. The proliferation capacity of CD44+/EPCAM high cells was higher than those of EPCAM low and LoVo cells, and the cell cycle of CD44+/EPCAM high cells was mostly in G0/G1 phase. Tumor formation rate of 500 CD44+/EPCAM high cells was 90% (9/10) in nude mice, with 1×104 EPCAM low cells being 0% (0/10). Moreover, expressions of CD44 and EPCAM in the second passage of CD44+/EPCAM high cells could still be detected. Conclusion: Colon cancer cell line LoVo contains CD44+/EPCAM high stem-like cells, and CD44+/EPCAM high cells can be used in further research of colon cancer stem cells.
2010, 17(2):155-160.
DOI: 10.3872/j.issn.1007-385X.2010.2.008
Abstract:
Abstract Objective:To explore the effect of Raf kinase inhibitor protein (RKIP) on the chemosensitivity of ovarian cancer SKOV-3 cells. Methods: Eukaryotic expression plasmid pcDNA3.1-ssRKIP containing full-length human RKIP cDNA was transfected into ovarian cancer cell line SKOV-3 by lipofect assay. Expression of RKIP in SKOV-3 cells was determined by Western blotting analysis. pcDNA3.1-ssRKIP-transfected SKOV-3 cells were treated with different concentrations of cisplatin, and the effect of RKIP on the proliferation of SKOV-3 cells treated with cisplatin was measured by MTS assay. Flow cytometry was used to detect the effect of RKIP on changes of apoptosis and cell cycle of SKOV-3 cells after cisplatin treatment. Results: The expression of RKIP in SKOV-3 cells was significantly increased after transfection with pcDNA3.1-ssRKIP. The growth inhibitory rate of SKOV-3 cells in pcDNA3.1-ssRKIP transfection group was significantly higher than that in the control group after treatment with different concentrations of cisplatin for 24 h, 48h or 72 h (P<0.05). After treatment with cisplatin at 2.5 μg/ml for 24 hours, pcDNA3.1-ssRKIP-transfected SKOV-3 cells showed a significantly higher percentage of apoptosis (10.86±0.73)% than non-transfected cells (4.27±0.67)% and empty vector-transfected cells (4.02±0.43)%. Without cisplatin treatment, the percentage of apoptosis for SKOV-3 cells transfected with pcDNA3.1-ssRKIP was (3.11±0.78)%, which was significantly higher than those of the non-transfected cells (1.51±0.13)% and empty vector-transfected cells (1.97±0.46)%. After cisplatin treatment, there were fewer cells in G0/G1 phase and more cells in S phase in pcDNA3.1-ssRKIP-transfected cells compared with the control cells, suggesting that cisplatin caused more S phase arrest in transfected cells. Conclusions: Over-expression of RKIP gene can increase chemosensitivity of ovarian cancer SKOV-3 cells to cisplatin.
Abstract Objective:To explore the effect of Raf kinase inhibitor protein (RKIP) on the chemosensitivity of ovarian cancer SKOV-3 cells. Methods: Eukaryotic expression plasmid pcDNA3.1-ssRKIP containing full-length human RKIP cDNA was transfected into ovarian cancer cell line SKOV-3 by lipofect assay. Expression of RKIP in SKOV-3 cells was determined by Western blotting analysis. pcDNA3.1-ssRKIP-transfected SKOV-3 cells were treated with different concentrations of cisplatin, and the effect of RKIP on the proliferation of SKOV-3 cells treated with cisplatin was measured by MTS assay. Flow cytometry was used to detect the effect of RKIP on changes of apoptosis and cell cycle of SKOV-3 cells after cisplatin treatment. Results: The expression of RKIP in SKOV-3 cells was significantly increased after transfection with pcDNA3.1-ssRKIP. The growth inhibitory rate of SKOV-3 cells in pcDNA3.1-ssRKIP transfection group was significantly higher than that in the control group after treatment with different concentrations of cisplatin for 24 h, 48h or 72 h (P<0.05). After treatment with cisplatin at 2.5 μg/ml for 24 hours, pcDNA3.1-ssRKIP-transfected SKOV-3 cells showed a significantly higher percentage of apoptosis (10.86±0.73)% than non-transfected cells (4.27±0.67)% and empty vector-transfected cells (4.02±0.43)%. Without cisplatin treatment, the percentage of apoptosis for SKOV-3 cells transfected with pcDNA3.1-ssRKIP was (3.11±0.78)%, which was significantly higher than those of the non-transfected cells (1.51±0.13)% and empty vector-transfected cells (1.97±0.46)%. After cisplatin treatment, there were fewer cells in G0/G1 phase and more cells in S phase in pcDNA3.1-ssRKIP-transfected cells compared with the control cells, suggesting that cisplatin caused more S phase arrest in transfected cells. Conclusions: Over-expression of RKIP gene can increase chemosensitivity of ovarian cancer SKOV-3 cells to cisplatin.
2010, 17(2):161-167.
DOI: 10.3872/j.issn.1007-385X.2010.2.009
Abstract:
Abstract Objective:To investigate the effect of antisense cathepsin B(CatB) RNA on the invasion and migration of human breast carcinoma cells. Methods: PBudCE4.1-antiCatB recombinant plasmid carrying antisense cathepsin B(CatB) gene was constructed and transfected into breast carcinoma cell line MDA-MB-231 by lipofection system. The expression of CatB protein in MDA-MB-231 cells was detected by Western blotting analysis; the proliferation of MDA-MB-231 cells was determined by MTT assay. Cell-matrix adhesion assay was used to examine the effect of anti-CatB on adhesion ability of MDA-MB-231 cells. The effects of anti-CatB on the invasion and migration abilities of MDA-MB-231 cells were measured by invasion and migration transwell system. Results: The recombinant plasmid PBudCE4.1-antiCatB was successfully constructed. Expression of CatB protein in MDA-MB-231 cells was decreased after PBudCE4.1-antiCatB transfection compared with those in untransfected and mock-vehicle transfected cells([0.96±0.02] vs [1.98±0.23], [1.84±0.08], P<0.05); the proliferation of MDA-MB-231 cells was also inhibited in PBudCE4.1-antiCatB transfected group([0.255±0.017] vs [0.458±0.033], [0.421±0.022], P<0.01); and the adhesion abilities(binding to matrix or fibronectin) were decreased([0.054±0.017] vs [0.111±0.018], [0.107±0.017], P<0.01; [0.052±0.008] vs [0.120±0.014], [0.113±0.009], P<0.01). Transwell asaay showed that the invasion and migration abilities were inhibited in PBudCE4.1-antiCatB transfected group compared with those in the non-transfection and mock-vehicle transfected groups([52.80±7.76] vs [124.00±44.54], [116.80±32.87], P<0.01; [60.25±8.73] vs [132.50±12.15], [119.20±25.13], P<0.01). Conclusion: The expression of antisense CatB RNA can inhibit the growth, adhesion, invasion and migration abilities of MDA-MB-231 cell in vitro.
Abstract Objective:To investigate the effect of antisense cathepsin B(CatB) RNA on the invasion and migration of human breast carcinoma cells. Methods: PBudCE4.1-antiCatB recombinant plasmid carrying antisense cathepsin B(CatB) gene was constructed and transfected into breast carcinoma cell line MDA-MB-231 by lipofection system. The expression of CatB protein in MDA-MB-231 cells was detected by Western blotting analysis; the proliferation of MDA-MB-231 cells was determined by MTT assay. Cell-matrix adhesion assay was used to examine the effect of anti-CatB on adhesion ability of MDA-MB-231 cells. The effects of anti-CatB on the invasion and migration abilities of MDA-MB-231 cells were measured by invasion and migration transwell system. Results: The recombinant plasmid PBudCE4.1-antiCatB was successfully constructed. Expression of CatB protein in MDA-MB-231 cells was decreased after PBudCE4.1-antiCatB transfection compared with those in untransfected and mock-vehicle transfected cells([0.96±0.02] vs [1.98±0.23], [1.84±0.08], P<0.05); the proliferation of MDA-MB-231 cells was also inhibited in PBudCE4.1-antiCatB transfected group([0.255±0.017] vs [0.458±0.033], [0.421±0.022], P<0.01); and the adhesion abilities(binding to matrix or fibronectin) were decreased([0.054±0.017] vs [0.111±0.018], [0.107±0.017], P<0.01; [0.052±0.008] vs [0.120±0.014], [0.113±0.009], P<0.01). Transwell asaay showed that the invasion and migration abilities were inhibited in PBudCE4.1-antiCatB transfected group compared with those in the non-transfection and mock-vehicle transfected groups([52.80±7.76] vs [124.00±44.54], [116.80±32.87], P<0.01; [60.25±8.73] vs [132.50±12.15], [119.20±25.13], P<0.01). Conclusion: The expression of antisense CatB RNA can inhibit the growth, adhesion, invasion and migration abilities of MDA-MB-231 cell in vitro.
2010, 17(2):168-173.
DOI: 10.3872/j.issn.1007-385X.2010.2.010
Abstract:
Abstract Objective:To investigate the effects of tumor metastasis-suppressor gene KiSS-1 on the invasion and migration abilities of osteosarcoma MG63 cells. Methods: KiSS-1 expression plasmid pSNAV2.0-KiSS-1 was constructed and transfected into MG63 cells. MG63 cells stably transfected with pSNAV2.0-KiSS-1 (named MG63-KiSS-1) were selected by G418. KiSS-1 mRNA and protein expression in MG63-KiSS-1 cells was examined by real-time PCR and Western blotting analysis, respectively. The invasion ability of MG63 cells was detected by transwell assay. The effects of KiSS-1 on the invasion and migration abilities of MG63 cells were measured by millicell assay and cell scratch healing assay. Results: MG63 cells stably transfected with pSNAV2.0-KiSS-1 were successfully established. MG63-KiSS-1 cells highly expressed KiSS-1 protein. The invasion ability of MG63 cells was significantly decreased after pSNAV2.0-KiSS-1 transfection (P<0.05). The migration ability of MG63 cells was also significantly inhibited after pSNAV2.0-KiSS-1 transfection as examined by millicell assay and cell scratch healing assay (P<0.05). Conclusion: KiSS-1 gene can significantly inhibit the invasion and migration abilities of osteosarcoma MG63 cells, which may play a key role in metastasis of osteosarcoma.
Abstract Objective:To investigate the effects of tumor metastasis-suppressor gene KiSS-1 on the invasion and migration abilities of osteosarcoma MG63 cells. Methods: KiSS-1 expression plasmid pSNAV2.0-KiSS-1 was constructed and transfected into MG63 cells. MG63 cells stably transfected with pSNAV2.0-KiSS-1 (named MG63-KiSS-1) were selected by G418. KiSS-1 mRNA and protein expression in MG63-KiSS-1 cells was examined by real-time PCR and Western blotting analysis, respectively. The invasion ability of MG63 cells was detected by transwell assay. The effects of KiSS-1 on the invasion and migration abilities of MG63 cells were measured by millicell assay and cell scratch healing assay. Results: MG63 cells stably transfected with pSNAV2.0-KiSS-1 were successfully established. MG63-KiSS-1 cells highly expressed KiSS-1 protein. The invasion ability of MG63 cells was significantly decreased after pSNAV2.0-KiSS-1 transfection (P<0.05). The migration ability of MG63 cells was also significantly inhibited after pSNAV2.0-KiSS-1 transfection as examined by millicell assay and cell scratch healing assay (P<0.05). Conclusion: KiSS-1 gene can significantly inhibit the invasion and migration abilities of osteosarcoma MG63 cells, which may play a key role in metastasis of osteosarcoma.
2010, 17(2):174-178.
DOI: 10.3872/j.issn.1007-385X.2010.2.011
Abstract:
Abstract Objective:To construct a recombinant eukaryotic expression vector mIL-21-pcDNA3.1 and transfect it into hepatoma cell line H22, so as to assess the biological activity of mIL-21. Methods: The gene fragment encoding mouse IL-21 was amplified by RT-PCR, and was then cloned into eukaryotic expression plasmid pcDNA3.1 to form recombinant plasmid mIL-21-pcDNA3.1. The recombinant plasmid is verified by DNA sequencing. mIL-21-pcDNA3.1 was transfected into H22 cells with lipofect regent, and its expression was detected by RT-PCR and Western blotting analysis. The effects of mIL-21-pcDNA3.1 on proliferation of T cells and cytotoxicity of NK cells were studied. Results: The recombinant plasmid mIL-21-pcDNA3.1 was confirmed by DNA sequencing. The expression of mIL-21 in H22 cells was confirmed by RT-PCR and Western blotting analysis. MTT results showed that stimulation index (SI) of T cells stimulated with mIL-2-H22 cell supernatant was 3.412±0.312, and the SI of ConA combination stimulating group was 4.673±0.450; both were significantly higher than those in the mock vector transfected (1.465±0.103) and untransfected groups (1.447±0.245, P<0.01). The cytotoxicity of NK cells in mIL-21-H22 cell supernatant group was (81.66±4.26)%, significantly higher than those in the mock vector transfected ([34.74±5.52]%) and untransfected groups ([33.61±1.42]%). Conclusion: The expression of mIL-21-pcDNA3.1 plasmid in H22 cells can enhance the proliferation of T cells and the cytotoxicity of NK cells, which lays a foundation for its role in the research of anti-hepatoma.
Abstract Objective:To construct a recombinant eukaryotic expression vector mIL-21-pcDNA3.1 and transfect it into hepatoma cell line H22, so as to assess the biological activity of mIL-21. Methods: The gene fragment encoding mouse IL-21 was amplified by RT-PCR, and was then cloned into eukaryotic expression plasmid pcDNA3.1 to form recombinant plasmid mIL-21-pcDNA3.1. The recombinant plasmid is verified by DNA sequencing. mIL-21-pcDNA3.1 was transfected into H22 cells with lipofect regent, and its expression was detected by RT-PCR and Western blotting analysis. The effects of mIL-21-pcDNA3.1 on proliferation of T cells and cytotoxicity of NK cells were studied. Results: The recombinant plasmid mIL-21-pcDNA3.1 was confirmed by DNA sequencing. The expression of mIL-21 in H22 cells was confirmed by RT-PCR and Western blotting analysis. MTT results showed that stimulation index (SI) of T cells stimulated with mIL-2-H22 cell supernatant was 3.412±0.312, and the SI of ConA combination stimulating group was 4.673±0.450; both were significantly higher than those in the mock vector transfected (1.465±0.103) and untransfected groups (1.447±0.245, P<0.01). The cytotoxicity of NK cells in mIL-21-H22 cell supernatant group was (81.66±4.26)%, significantly higher than those in the mock vector transfected ([34.74±5.52]%) and untransfected groups ([33.61±1.42]%). Conclusion: The expression of mIL-21-pcDNA3.1 plasmid in H22 cells can enhance the proliferation of T cells and the cytotoxicity of NK cells, which lays a foundation for its role in the research of anti-hepatoma.
2010, 17(2):179-182.
DOI: 10.3872/j.issn.1007-385X.2010.2.012
Abstract:
Abstract Objective: To investigate Foxp3 expression in neuroblastoma cell line SK-N-SH and its chemosensitivity to cyclophosvnamide (CTX) and pirarubicin (THP). Methods: SK-N-SH cells were cultured in vitro, and Foxp3 expression in SK-N-SH cells was examined by flow cytometry (FCM). The sensitive dosages of CTX and THP on SK-N-SH cells were determined by MTT assay. The effects of CTX or THP on Foxp3 expressions in SK-N-SH cells were examined by FCM and real-time PCR. Results: FCM results showed that SH-N-SK cells expressed high level of Foxp3. The sensitive dosage of CTX on SK-N-SH cells was 6 mmol/L, and that of THP was 80 ng/ml. CTX (6 mmol/L), THP (80 ng/ml) alone or in combination could not inhibit the expression of Foxp3 in SK-N-SH cells (P>0.05). Real-time PCR data also showed that CTX, THP alone or in combination could not down-regulate the expression of Foxp3 in SK-N-SH cells at mRNA level (P>0.05). Conclusion: Neuroblastoma SK-N-SH cells can express high level of Foxp3, but Foxp3 shows no chemosensitivity to CTX and THP.
Abstract Objective: To investigate Foxp3 expression in neuroblastoma cell line SK-N-SH and its chemosensitivity to cyclophosvnamide (CTX) and pirarubicin (THP). Methods: SK-N-SH cells were cultured in vitro, and Foxp3 expression in SK-N-SH cells was examined by flow cytometry (FCM). The sensitive dosages of CTX and THP on SK-N-SH cells were determined by MTT assay. The effects of CTX or THP on Foxp3 expressions in SK-N-SH cells were examined by FCM and real-time PCR. Results: FCM results showed that SH-N-SK cells expressed high level of Foxp3. The sensitive dosage of CTX on SK-N-SH cells was 6 mmol/L, and that of THP was 80 ng/ml. CTX (6 mmol/L), THP (80 ng/ml) alone or in combination could not inhibit the expression of Foxp3 in SK-N-SH cells (P>0.05). Real-time PCR data also showed that CTX, THP alone or in combination could not down-regulate the expression of Foxp3 in SK-N-SH cells at mRNA level (P>0.05). Conclusion: Neuroblastoma SK-N-SH cells can express high level of Foxp3, but Foxp3 shows no chemosensitivity to CTX and THP.
2010, 17(2):183-189.
DOI: 10.3872/j.issn.1007-385X.2010.2.013
Abstract:
Abstract Objective:To examine the expression of high mobility group A2 (HMGA2) mRNA in the peripheral blood of chronic myeloid leukemia (CML) patients and the related clinical characteristics, and to explore its role and clinical significance in CML progress. Methods: Peripheral blood samples of 24 CML patients and 5 volunteers, who had been diagnosed in our hospital from Jan. 2006 to Feb. 2008, were collected in the present study (all the patients and volunteers signed paper of informed consent and the study was approved by ethics committee). BCR/ABL fusion gene expression in CML bone samples was detected by fluorescence in situ hybridization (FISH). HMGA2 mRNA expression was examined by real-time fluorescence quantitative PCR (RTQ-PCR). Rank sum test was used to assess the HMGA2 gene transcription differences between CML-CP and CML-AP/BP patients. Correlation analyses were done using Spearman’s correlation test to explore the correlation between HMGA2 expression, BCR/ABL fusion gene expression, and hematological parameters in peripheral blood of CML patients. Results: We found that (58.08±39.21)% leukemia cells were BCR/ABL fusion gene-positive in 12 CML-CP patients, and the relative quantitative expression of HMGA2 gene was 2.39±1.86. Meanwhile, (87.50±16.21)% leukemia cells were BCR/ABL fusion gene-positive in 12 CML-AP/BP patients, and HMGA2 gene relative quantitative expression was 91.78±14.07. HMGA2 gene transcriptions between CML-CP and CML-AP/BP patients were significantly different (Z=-4.157, P<0.01). HMGA2 gene transcription in CML-AP/BP patients was positively correlated with the numbers of blast cells in the peripheral blood (r=0.636, P=0.017). Conclusion: HMGA2 gene transcription level in CML-AP/BP patients is higher than that in CML-CP patients, indicating HMGA2 may be a reliable indicator in estimating the development, prognosis and clinical treatment outcome of CML patients.
Abstract Objective:To examine the expression of high mobility group A2 (HMGA2) mRNA in the peripheral blood of chronic myeloid leukemia (CML) patients and the related clinical characteristics, and to explore its role and clinical significance in CML progress. Methods: Peripheral blood samples of 24 CML patients and 5 volunteers, who had been diagnosed in our hospital from Jan. 2006 to Feb. 2008, were collected in the present study (all the patients and volunteers signed paper of informed consent and the study was approved by ethics committee). BCR/ABL fusion gene expression in CML bone samples was detected by fluorescence in situ hybridization (FISH). HMGA2 mRNA expression was examined by real-time fluorescence quantitative PCR (RTQ-PCR). Rank sum test was used to assess the HMGA2 gene transcription differences between CML-CP and CML-AP/BP patients. Correlation analyses were done using Spearman’s correlation test to explore the correlation between HMGA2 expression, BCR/ABL fusion gene expression, and hematological parameters in peripheral blood of CML patients. Results: We found that (58.08±39.21)% leukemia cells were BCR/ABL fusion gene-positive in 12 CML-CP patients, and the relative quantitative expression of HMGA2 gene was 2.39±1.86. Meanwhile, (87.50±16.21)% leukemia cells were BCR/ABL fusion gene-positive in 12 CML-AP/BP patients, and HMGA2 gene relative quantitative expression was 91.78±14.07. HMGA2 gene transcriptions between CML-CP and CML-AP/BP patients were significantly different (Z=-4.157, P<0.01). HMGA2 gene transcription in CML-AP/BP patients was positively correlated with the numbers of blast cells in the peripheral blood (r=0.636, P=0.017). Conclusion: HMGA2 gene transcription level in CML-AP/BP patients is higher than that in CML-CP patients, indicating HMGA2 may be a reliable indicator in estimating the development, prognosis and clinical treatment outcome of CML patients.
2010, 17(2):190-193.
DOI: 10.3872/j.issn.1007-385X.2010.2.014
Abstract:
Abstract Objective:To investigate the expressions of cyclooxygenase-2(COX-2) and vascular endothelial growth factor-C(VEGF-C) in esophageal squamous cell carcinoma tissues and their correlation with tumor lymphangiogenesis. Methods: Totally 66 esophageal squamous cell carcinoma specimens were obtained from First Affiliated Hospital of Liaoning Medical University during Jan. 2002 to Jan. 2007, and 10 adjacent tissues were used as control. COX-2 and VEGF-C expressions in esophageal squamous cell carcinoma tissues were detected by SP immunohistochemistry. The lymphangiogenesis of tumor tissue and lymphatic vessel density (LVD) were determined by VEGFR-3 and type Ⅳ collagen immunohistochemical staining, and their relationship with lymph node metastasis was analzyed. Results: The positive rates of COX-2 and VEGF-C in 66 esophageal squamous cell carcinoma tissues (69.70% and 56.06%, respectively) were significantly higher than those in the adjacent tissues (P<0.05). The expressions of COX-2 and VEGF-C were remarkably higher in lymph node metastasis positive patients than those in metastasis negative patients (P<0.01). Positive correlation was observed between the expression of COX-2 and VEGF-C proteins (r=0.479, P<0.05). Significantly higher LVD was observed in COX-2 and VEGF-C double positive patients than in double negative patients (P<0.01). Conclusion: COX-2 and VEGF-C are highly expressed in esophageal squamous cell carcinoma tissues, and COX-2 might induce tumor lymphangiogenesis and lymph node metastasis by up-regulating VEGF-C expression.
Abstract Objective:To investigate the expressions of cyclooxygenase-2(COX-2) and vascular endothelial growth factor-C(VEGF-C) in esophageal squamous cell carcinoma tissues and their correlation with tumor lymphangiogenesis. Methods: Totally 66 esophageal squamous cell carcinoma specimens were obtained from First Affiliated Hospital of Liaoning Medical University during Jan. 2002 to Jan. 2007, and 10 adjacent tissues were used as control. COX-2 and VEGF-C expressions in esophageal squamous cell carcinoma tissues were detected by SP immunohistochemistry. The lymphangiogenesis of tumor tissue and lymphatic vessel density (LVD) were determined by VEGFR-3 and type Ⅳ collagen immunohistochemical staining, and their relationship with lymph node metastasis was analzyed. Results: The positive rates of COX-2 and VEGF-C in 66 esophageal squamous cell carcinoma tissues (69.70% and 56.06%, respectively) were significantly higher than those in the adjacent tissues (P<0.05). The expressions of COX-2 and VEGF-C were remarkably higher in lymph node metastasis positive patients than those in metastasis negative patients (P<0.01). Positive correlation was observed between the expression of COX-2 and VEGF-C proteins (r=0.479, P<0.05). Significantly higher LVD was observed in COX-2 and VEGF-C double positive patients than in double negative patients (P<0.01). Conclusion: COX-2 and VEGF-C are highly expressed in esophageal squamous cell carcinoma tissues, and COX-2 might induce tumor lymphangiogenesis and lymph node metastasis by up-regulating VEGF-C expression.
2010, 17(2):194-199.
DOI: 10.3872/j.issn.1007-385X.2010.2.015
Abstract:
Abstract Objective:To investigate expressions of iASPP and ASPP2 proteins in primary non-small cell lung cancer (NSCLC), and their relationship with P53 expression and the clinical characteristics of NSCLC. Methods: Specimens of 54 non-small cell lung cancer (NSCLC) patients, who were treated by surgery from Jan. to Dec. 2005 in Hospital of Guiyang Medical College, were included in this study. The expressions of iASPP, ASPP2 and P53 proteins were detected by immunohistochemistry staining and Western blotting analysis. iASPP and ASPP2 differential expressions in NSCLC tissues and adjacent cancer tissues and their relationship with P53 expression and clinical characteristics of NSCLC were analyzed. Results: The positive expression rate of iASPP in P53-negative NSCLC tissues was 71%, which was significantly higher than that in the 4 adjacent cancer tissues (21%, P=0.001). ASPP2 protein expression in NSCLC tissues was not significantly different from that in the adjacent lung tissues. iASPP expression in NSCLC tissues was significantly higher than that in the adjacent cancer tissues ( [0.57±0.36] vs [0.28±0.24], P=0.001). ASPP2 expression in NSCLC tissues was not significantly different from that in the adjacent lung tissues. iASPP and ASPP2 protein expressions in P53 positive NSCLC tissues were similar to those in the adjacent cancer tissues (P>0.05). iASPP protein expression in P53-negative NSCLC tissues was significantly higher than that in P53-positive NSCLC tissues (P<0.05). ASPP2 protein expression in P53-negative NSCLC tissues had no significant difference with that in P53-positive NSCLC tissues (P<0.05). iASPP and ASPP2 protein expressions in NSCLC patients were similar in patients of different genders, ages, pathology types, pathology grades, and clincial stages. Conclusion: iASPP expression is correlated with P53 expression in NSCLC tissues, with iASPP highly expressed in P53-negative NSCLC tissues. Expressions of iASPP and ASPP2 proteins have no correlation with the clinical characteristics of NSCLC.
Abstract Objective:To investigate expressions of iASPP and ASPP2 proteins in primary non-small cell lung cancer (NSCLC), and their relationship with P53 expression and the clinical characteristics of NSCLC. Methods: Specimens of 54 non-small cell lung cancer (NSCLC) patients, who were treated by surgery from Jan. to Dec. 2005 in Hospital of Guiyang Medical College, were included in this study. The expressions of iASPP, ASPP2 and P53 proteins were detected by immunohistochemistry staining and Western blotting analysis. iASPP and ASPP2 differential expressions in NSCLC tissues and adjacent cancer tissues and their relationship with P53 expression and clinical characteristics of NSCLC were analyzed. Results: The positive expression rate of iASPP in P53-negative NSCLC tissues was 71%, which was significantly higher than that in the 4 adjacent cancer tissues (21%, P=0.001). ASPP2 protein expression in NSCLC tissues was not significantly different from that in the adjacent lung tissues. iASPP expression in NSCLC tissues was significantly higher than that in the adjacent cancer tissues ( [0.57±0.36] vs [0.28±0.24], P=0.001). ASPP2 expression in NSCLC tissues was not significantly different from that in the adjacent lung tissues. iASPP and ASPP2 protein expressions in P53 positive NSCLC tissues were similar to those in the adjacent cancer tissues (P>0.05). iASPP protein expression in P53-negative NSCLC tissues was significantly higher than that in P53-positive NSCLC tissues (P<0.05). ASPP2 protein expression in P53-negative NSCLC tissues had no significant difference with that in P53-positive NSCLC tissues (P<0.05). iASPP and ASPP2 protein expressions in NSCLC patients were similar in patients of different genders, ages, pathology types, pathology grades, and clincial stages. Conclusion: iASPP expression is correlated with P53 expression in NSCLC tissues, with iASPP highly expressed in P53-negative NSCLC tissues. Expressions of iASPP and ASPP2 proteins have no correlation with the clinical characteristics of NSCLC.
2010, 17(2):200-204.
DOI: 10.3872/j.issn.1007-385X.2010.2.016
Abstract:
Abstract Objective: To investigate the expression of Rho guanine nucleotide dissociation inhibitor(RhoGDI) protein in lung cancer and to evaluate its clinical significance. Methods: The expression of RhoGDI protein in 39 lung cancer specimens and 5 lung inflammatory pseudotumor tissues were detected by Western blotting analysis. The tissues were collected from patients treated in the two affiliated hospitals of Soochow University during 2007-2008. The relationship between RhoGDI expression and the clinical and pathological characteristics of lung cancer were analyzed. Results: RhoGDI protein level in small-cell lung cancer(SCLC) tissues was significantly higher than those in non-small cell lung cancer (NSCLC) and inflammatory pseudotumor tissues ([0.903±0.228] vs [0.522±0.152], [0.485±0.095], P<0.05). RhoGDI protein level was significantly higher in poorly-differentiated tumors than those in moderately-, highly-differentiated NSCLC and inflammatory pseudotumor tissues ([0.649±0.123] vs [0.458±0.138], [0.485±0.095], P<0.05). RhoGDI protein level in squamous carcinoma had no difference with that in adenocarcinoma. The level of RhoGDI protein in NSCLC tissues was increased gradually when the pathology of tumor stage increased (Ⅰ-Ⅲ), but with no significant difference was found among them (F=0.6, P>0.05). RhoGDI protein level in lung cancer was not related to the tumor size (t=1.15, P>0.05) or lymphatic metastasis (t=1.648, P>0.05). RhoGDI protein expression in NSCLC was positively correlated with the serum levels of lactate dehydrogenate (r=0.46, P＜0.01). Conclusion: RhoGDI protein expression is correlated with pathological characteristics and differentiation stages of lung cancer.
Abstract Objective: To investigate the expression of Rho guanine nucleotide dissociation inhibitor(RhoGDI) protein in lung cancer and to evaluate its clinical significance. Methods: The expression of RhoGDI protein in 39 lung cancer specimens and 5 lung inflammatory pseudotumor tissues were detected by Western blotting analysis. The tissues were collected from patients treated in the two affiliated hospitals of Soochow University during 2007-2008. The relationship between RhoGDI expression and the clinical and pathological characteristics of lung cancer were analyzed. Results: RhoGDI protein level in small-cell lung cancer(SCLC) tissues was significantly higher than those in non-small cell lung cancer (NSCLC) and inflammatory pseudotumor tissues ([0.903±0.228] vs [0.522±0.152], [0.485±0.095], P<0.05). RhoGDI protein level was significantly higher in poorly-differentiated tumors than those in moderately-, highly-differentiated NSCLC and inflammatory pseudotumor tissues ([0.649±0.123] vs [0.458±0.138], [0.485±0.095], P<0.05). RhoGDI protein level in squamous carcinoma had no difference with that in adenocarcinoma. The level of RhoGDI protein in NSCLC tissues was increased gradually when the pathology of tumor stage increased (Ⅰ-Ⅲ), but with no significant difference was found among them (F=0.6, P>0.05). RhoGDI protein level in lung cancer was not related to the tumor size (t=1.15, P>0.05) or lymphatic metastasis (t=1.648, P>0.05). RhoGDI protein expression in NSCLC was positively correlated with the serum levels of lactate dehydrogenate (r=0.46, P＜0.01). Conclusion: RhoGDI protein expression is correlated with pathological characteristics and differentiation stages of lung cancer.
17 Expressions of Ki-67 and LRP in different subtypes of breast cancer and their clinical significance
2010, 17(2):205-209.
DOI: 10.3872/j.issn.1007-385X.2010.2.017
Abstract:
Abstract Objective:To investigate the expressions of Ki-67 and LRP in different breast cancer subtypes and the relevant clinical significance. Methods: Breast cancer specimens were collected from 203 patients who underwent operation in the Affiliated Tumor Hospital of Xinjiang Medical University during Jan. to Oct. 2009. The expression of ER, PR, HER2, Ki-67 and LRP in the breast cancer tissues was determined by immunohistochemical staining. Ki-67 and LRP expression and its correlation with clinical and pathological features of different breast cancer subtypes was analyzed. Results: Four breast cancer subtypes were identified with different ER, PR and HER2 gene expression patterns. There were significant differences in tumor sizes, clinical stages, histological grades, lymph node metastases, and patient’ ages between the 4 breast cancer subtypes(P<0.05), but not in cancer type. Ki-67 and LRP proteins were highly expressed in Luminal B(ER/PR+, HER2+) breast cancer(93.2%, 86.2%, P<0.05), but the expression of the two was not correlated with each other(r=0.144, P>0.05). The chemotherapy efficiency for LRP-positive Luminal B patients was lower than that in LRP-negative Luminal B patients(39.4% vs 83.3%, P<0.05), while the chemotherapy efficiencies between Ki-67-positive and negative Luminal B patients were not significantly different(P>0.05). Conclusion: Expression of Ki-67 and LRP proteins varies in different breast cancer subtypes. Overexpression of LRP in Luminal B breast cancer might be correlated with the chemotherapy efficiency after surgery.
Abstract Objective:To investigate the expressions of Ki-67 and LRP in different breast cancer subtypes and the relevant clinical significance. Methods: Breast cancer specimens were collected from 203 patients who underwent operation in the Affiliated Tumor Hospital of Xinjiang Medical University during Jan. to Oct. 2009. The expression of ER, PR, HER2, Ki-67 and LRP in the breast cancer tissues was determined by immunohistochemical staining. Ki-67 and LRP expression and its correlation with clinical and pathological features of different breast cancer subtypes was analyzed. Results: Four breast cancer subtypes were identified with different ER, PR and HER2 gene expression patterns. There were significant differences in tumor sizes, clinical stages, histological grades, lymph node metastases, and patient’ ages between the 4 breast cancer subtypes(P<0.05), but not in cancer type. Ki-67 and LRP proteins were highly expressed in Luminal B(ER/PR+, HER2+) breast cancer(93.2%, 86.2%, P<0.05), but the expression of the two was not correlated with each other(r=0.144, P>0.05). The chemotherapy efficiency for LRP-positive Luminal B patients was lower than that in LRP-negative Luminal B patients(39.4% vs 83.3%, P<0.05), while the chemotherapy efficiencies between Ki-67-positive and negative Luminal B patients were not significantly different(P>0.05). Conclusion: Expression of Ki-67 and LRP proteins varies in different breast cancer subtypes. Overexpression of LRP in Luminal B breast cancer might be correlated with the chemotherapy efficiency after surgery.
18 Role of integrin α5β1-mediated PI3K-AKT signal pathway in pathologenesis of chronic myeloid leukemia
2010, 17(2):210-212.
DOI: 10.3872/j.issn.1007-385X.2010.2.018
Abstract:
摘 要 目的：研究整合素α5β1介导的PI3K-AKT信号转导通路在慢性髓细胞白血病（chronic myeloid leukemia，CML）发病机制中的作用。方法：流式细胞术检测不同剂量抗整合素α5β1单抗（Anti-α5β1）作用后白血病K562细胞的凋亡率， Western blotting检测健康志愿者、CML急变期患者、K562细胞和Anti-α5β1处理的K562细胞中PI3K和AKT蛋白的表达水平。结果：Anti-α5β1可诱导K562细胞凋亡，并呈剂量依赖性。与健康志愿者比较，CML急变期患者骨髓单个核细胞和K562细胞中PI3K和AKT蛋白的表达水平明显增高（P<0.05）；Anti-α5β1处理后，K562细胞内PI3K、AKT蛋白表达水平明显降低，其磷酸化水平也下降（P<0.05）。结论: 整合素α5β1可能通过影响PI3K-AKT信号转导通路中关键分子PI3K和AKT的表达水平诱导CML祖细胞凋亡耐受，参与了CML的发生、发展过程。
摘 要 目的：研究整合素α5β1介导的PI3K-AKT信号转导通路在慢性髓细胞白血病（chronic myeloid leukemia，CML）发病机制中的作用。方法：流式细胞术检测不同剂量抗整合素α5β1单抗（Anti-α5β1）作用后白血病K562细胞的凋亡率， Western blotting检测健康志愿者、CML急变期患者、K562细胞和Anti-α5β1处理的K562细胞中PI3K和AKT蛋白的表达水平。结果：Anti-α5β1可诱导K562细胞凋亡，并呈剂量依赖性。与健康志愿者比较，CML急变期患者骨髓单个核细胞和K562细胞中PI3K和AKT蛋白的表达水平明显增高（P<0.05）；Anti-α5β1处理后，K562细胞内PI3K、AKT蛋白表达水平明显降低，其磷酸化水平也下降（P<0.05）。结论: 整合素α5β1可能通过影响PI3K-AKT信号转导通路中关键分子PI3K和AKT的表达水平诱导CML祖细胞凋亡耐受，参与了CML的发生、发展过程。
2010, 17(2):213-220.
DOI: 10.3872/j.issn.1007-385X.2010.2.019
Abstract:
Abstract Tumor is a chronic disease caused by accumulation of gene mutation, but the development and progression of certain tumors is dependent on one or a few genes, which is called oncogene addiction or oncogene dependent phenomenon. Weinstein first described this phenomenon as ‘oncogene addiction’ early in 2002, and to date there have been 3 models to explain this phenomenon, namely, the ‘bizarre circuitry model’, ‘oncogenic shock model’, and ‘oncogenic selection model’. The complexity of cancers and heterogeneity of cancer patients set great barriers for identifying the ‘additive oncogene’. Here we introduce the high-throughput screening methods for identifying the state of oncogene addiction, including genomics, high-throughput proteomics, gene knockout system, and the emerging RNA interference. The ‘oncogene addiction’ theory can well explain the mechanism of target therapy and enforce the evidence for development of molecular target agents. ‘Oncogene addiction’ theory has been viewed as of great potential for cancer target therapy, although it faces some challenges and needs further research.
Abstract Tumor is a chronic disease caused by accumulation of gene mutation, but the development and progression of certain tumors is dependent on one or a few genes, which is called oncogene addiction or oncogene dependent phenomenon. Weinstein first described this phenomenon as ‘oncogene addiction’ early in 2002, and to date there have been 3 models to explain this phenomenon, namely, the ‘bizarre circuitry model’, ‘oncogenic shock model’, and ‘oncogenic selection model’. The complexity of cancers and heterogeneity of cancer patients set great barriers for identifying the ‘additive oncogene’. Here we introduce the high-throughput screening methods for identifying the state of oncogene addiction, including genomics, high-throughput proteomics, gene knockout system, and the emerging RNA interference. The ‘oncogene addiction’ theory can well explain the mechanism of target therapy and enforce the evidence for development of molecular target agents. ‘Oncogene addiction’ theory has been viewed as of great potential for cancer target therapy, although it faces some challenges and needs further research.
2010, 17(2):221-226.
DOI: 10.3872/j.issn.1007-385X.2010.2.020
Abstract:
Abstract Overexpression of HER-2/neu oncogene is a frequent molecular event in multiple human cancers, including in approximately 30% of all primary breast cancer case. The humanized anti-HER-2 monoclonal antibody, trastuzumab (also named as herceptin), has been proven to be effective in patients with HER-2-associated metastatic breast cancer since 1990s. Great progress has been made in developing various vaccines targeting HER-2, including peptide vaccines, protein vaccines, cell vaccines, dendritic cell-associated vaccines, and DNA vaccines. For example, peptide E75 (p369-399) combined with granulocyte-macrophage colony-stimulating factor (GM-CSF) has been shown effective for HLA-A2(+)/A3(+) stage Ⅱ breast cancer patients, and HER-2-targeted DNA vaccine has already entered clinical trials. They might be important ways for treatment of breast cancer. Anyhow, there are still a lot of problems need to be addressed before their application in clinical practice.
Abstract Overexpression of HER-2/neu oncogene is a frequent molecular event in multiple human cancers, including in approximately 30% of all primary breast cancer case. The humanized anti-HER-2 monoclonal antibody, trastuzumab (also named as herceptin), has been proven to be effective in patients with HER-2-associated metastatic breast cancer since 1990s. Great progress has been made in developing various vaccines targeting HER-2, including peptide vaccines, protein vaccines, cell vaccines, dendritic cell-associated vaccines, and DNA vaccines. For example, peptide E75 (p369-399) combined with granulocyte-macrophage colony-stimulating factor (GM-CSF) has been shown effective for HLA-A2(+)/A3(+) stage Ⅱ breast cancer patients, and HER-2-targeted DNA vaccine has already entered clinical trials. They might be important ways for treatment of breast cancer. Anyhow, there are still a lot of problems need to be addressed before their application in clinical practice.
2010, 17(2):227-231.
DOI: 10.3872/j.issn.1007-385X.2010.2.021
Abstract:
Abstract Breast cancer metastasis is a complicated process driven by multiple genes and follows multiple steps. The regulation of metastasis-associated genes is the molecular basis for metastasis. Metastasis-associated genes can promote or suppress the metastasis potential of tumor cells without affecting their growth and proliferation. Generally, metastasis-associated genes fall into two categories: metastasis-promoter genes and metastasis-suppressor genes, including oncogenes, tumor suppressor genes, signaling molecule genes, adhesion molecule genes, cytokine genes, matrix metalloproteinase genes, etc. The elucidation of the underlying mechanisms of breast cancer metastasis-associated genes and the related downstream signaling pathways can facilitate the molecular diagnosis and individualized therapy of patients with metastasic breast cancers.
Abstract Breast cancer metastasis is a complicated process driven by multiple genes and follows multiple steps. The regulation of metastasis-associated genes is the molecular basis for metastasis. Metastasis-associated genes can promote or suppress the metastasis potential of tumor cells without affecting their growth and proliferation. Generally, metastasis-associated genes fall into two categories: metastasis-promoter genes and metastasis-suppressor genes, including oncogenes, tumor suppressor genes, signaling molecule genes, adhesion molecule genes, cytokine genes, matrix metalloproteinase genes, etc. The elucidation of the underlying mechanisms of breast cancer metastasis-associated genes and the related downstream signaling pathways can facilitate the molecular diagnosis and individualized therapy of patients with metastasic breast cancers.
2010, 17(2):232-236.
DOI: 10.3872/j.issn.1007-385X.2010.2.022
Abstract:
Abstract Tumor cells make compensation for ischemia and hypoxia through glucose transport, enhancement of glycolysis, and formation of tumor neoangiogenesis system. And glucose uptake is an important way to increase energy intake. Glucose transporter 1 (GLUT-1) is an important carrier of glucose transmembrane transport for tissue cells, and it is lowly expressed in mammalian embryos and mature tissues, but highly expressed in malignant tumor cells in a hypoxia and ischemia microenvironment. Expression of GLUT-1 is associated with tumor progression and prognosis. The regulation of GLUT-1 can be divided into acute and chronic pathways in tumor cells in an in vitro culture system. Chronic pathway concerning hypoxia-inducible factor-1 mRNA and protein syntheses is the main regulation pathway. Detection GLUT-1 expression in tumor tissues by immunohistochemistry and RT-PCR assay may provide a new way for tumor diagnosis. Blocking the fundamental energy source of tumor by GLUT-1-targeted therapy may provide a new strategy for cancer treatment.
Abstract Tumor cells make compensation for ischemia and hypoxia through glucose transport, enhancement of glycolysis, and formation of tumor neoangiogenesis system. And glucose uptake is an important way to increase energy intake. Glucose transporter 1 (GLUT-1) is an important carrier of glucose transmembrane transport for tissue cells, and it is lowly expressed in mammalian embryos and mature tissues, but highly expressed in malignant tumor cells in a hypoxia and ischemia microenvironment. Expression of GLUT-1 is associated with tumor progression and prognosis. The regulation of GLUT-1 can be divided into acute and chronic pathways in tumor cells in an in vitro culture system. Chronic pathway concerning hypoxia-inducible factor-1 mRNA and protein syntheses is the main regulation pathway. Detection GLUT-1 expression in tumor tissues by immunohistochemistry and RT-PCR assay may provide a new way for tumor diagnosis. Blocking the fundamental energy source of tumor by GLUT-1-targeted therapy may provide a new strategy for cancer treatment.
2010, 17(2):237-241.
DOI: 10.3872/j.issn.1007-385X.2010.2.023
Abstract:
Abstract Estrogen receptors (ER), including estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ), are ligand-dependent trans-acting transcription factors of steroid hormone nuclear receptor family. They have three major functional domains: the N-terminal region, DNA-binding domain and the C-terminal region. ERα and ERβ are expressed in non-small cell lung cancer (NSCLC) tissues and normal lung tissues, and their expressions are correlated with histological types of lung cancer. ER regulates transcription through the estrogen signaling pathway, and regulates its active form in the nuclei of tumor cells through the growth factor receptor pathway and steroid signaling pathway, thereby affecting the biological behaviors of cells such as growth, division, and metabolism. The hypermethylation of ER is related to the occurrence of lung tumors. ERα does not affect the metabolism of tobacco carcinogens though the correlation of smoking, and lung cancer is more evident in women. Whether the expression of ERα or ERβ is a valid predictor of lung cancer needs to be studied further. In conclusion, ERα and ERβ are closely correlated with the development, progression，and prognosis of NSCLC. Hormonal therapy based on ERα and ERβ may become an important strategy for the treatment of lung cancer.
Abstract Estrogen receptors (ER), including estrogen receptor alpha (ERα) and estrogen receptor beta (ERβ), are ligand-dependent trans-acting transcription factors of steroid hormone nuclear receptor family. They have three major functional domains: the N-terminal region, DNA-binding domain and the C-terminal region. ERα and ERβ are expressed in non-small cell lung cancer (NSCLC) tissues and normal lung tissues, and their expressions are correlated with histological types of lung cancer. ER regulates transcription through the estrogen signaling pathway, and regulates its active form in the nuclei of tumor cells through the growth factor receptor pathway and steroid signaling pathway, thereby affecting the biological behaviors of cells such as growth, division, and metabolism. The hypermethylation of ER is related to the occurrence of lung tumors. ERα does not affect the metabolism of tobacco carcinogens though the correlation of smoking, and lung cancer is more evident in women. Whether the expression of ERα or ERβ is a valid predictor of lung cancer needs to be studied further. In conclusion, ERα and ERβ are closely correlated with the development, progression，and prognosis of NSCLC. Hormonal therapy based on ERα and ERβ may become an important strategy for the treatment of lung cancer.
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- 《中国肿瘤生物治疗杂志》
- 1994年创刊
- 主办单位：Chinese Society of Immunology, Chinese Anti-cancer Association
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- 刊号：ISSN 1007-385X
- CN 31-1725/R
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
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Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.