Mobile website
Volume 22,Issue 1,2015 Table of Contents
2015, 22(1):1-7.
DOI: 10.3872/j.issn.1007-385X.2015.1.001
Abstract:
MicroRNAs plays critical roles in diverse physiological and pathological processes. It has attracted worldwide attention and rapid progress has been achieved in the past two decades. Hepatocellular carcinoma (HCC), especially hepatitis B virus (HBV)-related HCC, is particularly prevalent in China. Recently, as the possible roles for microRNAs in the pathogenesis and progression of HCC is being gradually uncovered, significant advances have been made in microRNA-based diagnosis and therapy of HCC. Owing to the rapid progress of new techniques such as massive parallel signature sequencing, miRNomes of human normal liver and HCC have been revealed, then deepened our understanding of the important microRNAs in HCC development. The enriched microRNAs in the liver are reported to be deregulated during HCC development, and these microRNAs are suggested to bear considerable therapeutic potential in HCC gene therapy. Moreover, serum microRNAs in HCC patients are also determined to be biomarkers for HCC diagnosis and individual therapy. This review paper aims to summarize the recent literature on the regulation of microRNAs expression in HCC, downstream targets of microRNAs in HCC development, and outlook the potential of serum microRNAs as diagnostic/prognostic makers and therapeutic targets for HCC.
MicroRNAs plays critical roles in diverse physiological and pathological processes. It has attracted worldwide attention and rapid progress has been achieved in the past two decades. Hepatocellular carcinoma (HCC), especially hepatitis B virus (HBV)-related HCC, is particularly prevalent in China. Recently, as the possible roles for microRNAs in the pathogenesis and progression of HCC is being gradually uncovered, significant advances have been made in microRNA-based diagnosis and therapy of HCC. Owing to the rapid progress of new techniques such as massive parallel signature sequencing, miRNomes of human normal liver and HCC have been revealed, then deepened our understanding of the important microRNAs in HCC development. The enriched microRNAs in the liver are reported to be deregulated during HCC development, and these microRNAs are suggested to bear considerable therapeutic potential in HCC gene therapy. Moreover, serum microRNAs in HCC patients are also determined to be biomarkers for HCC diagnosis and individual therapy. This review paper aims to summarize the recent literature on the regulation of microRNAs expression in HCC, downstream targets of microRNAs in HCC development, and outlook the potential of serum microRNAs as diagnostic/prognostic makers and therapeutic targets for HCC.
2015, 22(1):8-15.
DOI: 10.3872/j.issn.1007-385X.2015.1.002
Abstract:
Among the various available cancer treatment modalities, anti-tumor immune escape is attracting tremendous attention worldwide and has become one of the fast-developing research fields, especially when the anti-prostate cancer vaccine was approved by the FDA and the monocloan antibodies, which targeted PD-1 and CTLA-4, were successfully launched into market. Since 2010, tremendous research progress has been made on cellular immunotherapy of cancer in China. Nevertheless, practically cellular immunotherapy of cancer is facing several challenging issues. Including: 1) how to choose the proper treatment modality and time duration; 2) how to manage the quality control; 3) how to evaluate the treatment outcome; and 4) how to obtain stronger empirical evidence to support the benefits of the treatment. With regard to quality management and outcome evaluation, there exist similarities and differences between cellular immunotherapy and traditional biological and chemical drugs for cancers. Similar to the quality control of biological and chemical treatments of cancer, the quality management of cellular immunotherapy of cancer involves such aspects as the structure of the clinic or center, personnel training and testing program. In contrast, the outcome evaluation of cellular immunotherapy of cancer cannot simply follow the traditional method for biological and chemical cancer therapies. This paper aims to review the recent development in the quality management and outcome evaluation of cellular immunotherapy of cancer, hoping to bring these issues to the attention of peers in China.
Among the various available cancer treatment modalities, anti-tumor immune escape is attracting tremendous attention worldwide and has become one of the fast-developing research fields, especially when the anti-prostate cancer vaccine was approved by the FDA and the monocloan antibodies, which targeted PD-1 and CTLA-4, were successfully launched into market. Since 2010, tremendous research progress has been made on cellular immunotherapy of cancer in China. Nevertheless, practically cellular immunotherapy of cancer is facing several challenging issues. Including: 1) how to choose the proper treatment modality and time duration; 2) how to manage the quality control; 3) how to evaluate the treatment outcome; and 4) how to obtain stronger empirical evidence to support the benefits of the treatment. With regard to quality management and outcome evaluation, there exist similarities and differences between cellular immunotherapy and traditional biological and chemical drugs for cancers. Similar to the quality control of biological and chemical treatments of cancer, the quality management of cellular immunotherapy of cancer involves such aspects as the structure of the clinic or center, personnel training and testing program. In contrast, the outcome evaluation of cellular immunotherapy of cancer cannot simply follow the traditional method for biological and chemical cancer therapies. This paper aims to review the recent development in the quality management and outcome evaluation of cellular immunotherapy of cancer, hoping to bring these issues to the attention of peers in China.
2015, 22(1):16-21.
DOI: 10.3872/j.issn.1007-385X.2015.1.003
Abstract:
Objective : To investigate the anti-tumor effects of a dual cancer-specific oncolytic adenovirus Ad-Apoptin-PEG3p-E1a on colorectal cancer in vitro and in vivo. Methods: A dual cancer-specific oncolytic adenovirus vector carried Apoptin and PEG3 promoter (Ad-Apoptin-PEG3p-E1a) was constructed. In experiments in vitro, colon cancer SW1116 cells and gastric epithelial GES cells were infected with the constructed adenoviral vector. At various time points after infection, cell viability was assessed by MTT assay, apoptosis, levels of reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and cytochrome C (Cyt C) by flow cytometry. In experiments in vivo, BALB/c mice bearing established subcutaneous transplantation tumors were treated with Ad-Apoptin-PEG3p-E1a and a control vector respectively. Tumor growth kinetics and mean survival time were assessed. Results: In vitro, Ad-Apoptin-PEG3p-E1a significantly suppressed proliferation, increased ROS production, reduced MMP levels and enhanced Cyt C release in a dose- and time-dependent manner in SW1116 cells, as compared with the control vector. Ad-Apoptin-PEG3p-E1a infection also resulted in increased apoptosis in SW1116 cells but had no significant effects on GES cells. In vivo, Ad-Apoptin-PEG3p-E1a inhibited the growth of subcutaneous transplantation tumors significantly and extended the lifespan of the animals (41.0±07 days), while Ad-PEG3p-E1a-, Ad-Apoptin- and Ad-mock-treated animals had shorter mean survival time (34.4±16 days, 33.2±1.2 and 28.4±1.4, respectively, P<0.05）. Conclusion: The dual cancer-specific oncolytic adenovirus vector carrying Apoptin and PEG3p may offer a promising gene therapy for colon cancer.
Objective : To investigate the anti-tumor effects of a dual cancer-specific oncolytic adenovirus Ad-Apoptin-PEG3p-E1a on colorectal cancer in vitro and in vivo. Methods: A dual cancer-specific oncolytic adenovirus vector carried Apoptin and PEG3 promoter (Ad-Apoptin-PEG3p-E1a) was constructed. In experiments in vitro, colon cancer SW1116 cells and gastric epithelial GES cells were infected with the constructed adenoviral vector. At various time points after infection, cell viability was assessed by MTT assay, apoptosis, levels of reactive oxygen species (ROS), mitochondrial membrane potential (MMP) and cytochrome C (Cyt C) by flow cytometry. In experiments in vivo, BALB/c mice bearing established subcutaneous transplantation tumors were treated with Ad-Apoptin-PEG3p-E1a and a control vector respectively. Tumor growth kinetics and mean survival time were assessed. Results: In vitro, Ad-Apoptin-PEG3p-E1a significantly suppressed proliferation, increased ROS production, reduced MMP levels and enhanced Cyt C release in a dose- and time-dependent manner in SW1116 cells, as compared with the control vector. Ad-Apoptin-PEG3p-E1a infection also resulted in increased apoptosis in SW1116 cells but had no significant effects on GES cells. In vivo, Ad-Apoptin-PEG3p-E1a inhibited the growth of subcutaneous transplantation tumors significantly and extended the lifespan of the animals (41.0±07 days), while Ad-PEG3p-E1a-, Ad-Apoptin- and Ad-mock-treated animals had shorter mean survival time (34.4±16 days, 33.2±1.2 and 28.4±1.4, respectively, P<0.05）. Conclusion: The dual cancer-specific oncolytic adenovirus vector carrying Apoptin and PEG3p may offer a promising gene therapy for colon cancer.
2015, 22(1):22-27.
DOI: 10.3872/j.issn.1007-385X.2015.1.004
Abstract:
Objectiv : To study the anti-tumor activity and mechanism of Bursal-derived pentapeptide-Ⅱ(BPP-Ⅱ). Methods: Tumor cells including murine B-cell lymphoma WEHI-231 cells, human nasopharyngeal carcinoma CNE cells and rat hepatoma RH-35 cells and normal cells including human embryonic kidney 293 cells, pig kidney PK15 cells and Chinese hamster ovary CHO cells were stimulated with BPP-Ⅱ at 0.02 μg/ml, 0.2 μg/ml, 2 μg/ml and 20 μg/ml for 48 h. Cell viability was then measured by MTT assays, p53 luciferase activity and the expression of P53 at the protein level were assessed, and the effect of BPP-Ⅱ binding peptides biopaned from a phage display 12-mer random peptide library on BPP-Ⅱ-mediated inhibition of WEHI-231 cell proliferation was determined by MTT assays. Results: BPP-Ⅱ inhibited the proliferation of tumor cells but not normal cells, activated p53 transcription, and increased P53 protein content. After four rounds of biopanning, three BPP-Ⅱ binding peptides were obtained: QSLPSPLWIQQS (P3-12), DRMPDSAWTTRK (P5-12) and ALWPPNLHAWVP (P6-12). P3-12 significantly inhibited the anti-proliferative activity of BPP-Ⅱ at both 2 μg/ml (97.5±3.4)% and 20 μg/ml (98.9±3.5)% as compared with the control (86.3±1.9)% in WEHI-231 cells (P<0.05). At 20 μg/ml, both P5-12 (96.7±3.1)% and P6-12 (95.4±3.8)% inhibited the anti-proliferative activity of BPP-Ⅱ as compared with the control (86.3±1.9%) in WEHI-231 cells (P<0.05). Conclusions: BPP-Ⅱ appears to have an antitumor activity, possibly attributable to p53 activation. Taken together, our findings suggest a significant clinical importance for BPP-Ⅱ from the perspective of cancer therapy.
Objectiv : To study the anti-tumor activity and mechanism of Bursal-derived pentapeptide-Ⅱ(BPP-Ⅱ). Methods: Tumor cells including murine B-cell lymphoma WEHI-231 cells, human nasopharyngeal carcinoma CNE cells and rat hepatoma RH-35 cells and normal cells including human embryonic kidney 293 cells, pig kidney PK15 cells and Chinese hamster ovary CHO cells were stimulated with BPP-Ⅱ at 0.02 μg/ml, 0.2 μg/ml, 2 μg/ml and 20 μg/ml for 48 h. Cell viability was then measured by MTT assays, p53 luciferase activity and the expression of P53 at the protein level were assessed, and the effect of BPP-Ⅱ binding peptides biopaned from a phage display 12-mer random peptide library on BPP-Ⅱ-mediated inhibition of WEHI-231 cell proliferation was determined by MTT assays. Results: BPP-Ⅱ inhibited the proliferation of tumor cells but not normal cells, activated p53 transcription, and increased P53 protein content. After four rounds of biopanning, three BPP-Ⅱ binding peptides were obtained: QSLPSPLWIQQS (P3-12), DRMPDSAWTTRK (P5-12) and ALWPPNLHAWVP (P6-12). P3-12 significantly inhibited the anti-proliferative activity of BPP-Ⅱ at both 2 μg/ml (97.5±3.4)% and 20 μg/ml (98.9±3.5)% as compared with the control (86.3±1.9)% in WEHI-231 cells (P<0.05). At 20 μg/ml, both P5-12 (96.7±3.1)% and P6-12 (95.4±3.8)% inhibited the anti-proliferative activity of BPP-Ⅱ as compared with the control (86.3±1.9%) in WEHI-231 cells (P<0.05). Conclusions: BPP-Ⅱ appears to have an antitumor activity, possibly attributable to p53 activation. Taken together, our findings suggest a significant clinical importance for BPP-Ⅱ from the perspective of cancer therapy.
2015, 22(1):28-33.
DOI: 10.3872/j.issn.1007-385X.2015.1.005
Abstract:
Objective : To determine the expression and possible roles of miR-29a and B7-H3 in glioma growth and invasion. Methods: Glioma tissue specimens were collected from 19 patients who underwent surgical glioma resection in the Department of Neurosurgery, Soochow University-Affiliated First Hospital between September, 2006 and December, 2010. Levels of miR-29a and B7-H3 mRNAs in the tissue specimens and human glioma U87 cells were determined by Real-time PCR. U87 cells were transfected with miR-29a mimics, and B7-H3 expression and invasive capacity in the transfectants were assessed by flow cytometry and transwell migration assay, respectively. The expression of invasion-related chemical chemokines was analyzed by flow cytometry, and the ability of miR-29a to bind to CXCR4 was predicted by the miRtarbase software. Results: In glioma tissue specimens, miR-29a and B7-H3 mRNA levels were inversely correlated. In vitro, miR-29a down-regulated B7-H3 mRNAs expression in U87 cells and miR-29a-mediated down-regulation of B7-H3 resulted in a significant decrease in the invasive ability but not proliferative activity in U87 cells. Parallel to down-regulation of B7-H3 expression, CXCR4 expression was also down-regulated in U87 cells transfected with miR-29a mimics. No miR-29a binding sites were detected in the CXCR4 gene. Conclusions: In human glioma, miR-29a can effectively down-regulate B7-H3 expression and inhibit invasive activity. It is likely that these effects of miR-29a were at least attributable dwon-regulated expression of CXCR4.
Objective : To determine the expression and possible roles of miR-29a and B7-H3 in glioma growth and invasion. Methods: Glioma tissue specimens were collected from 19 patients who underwent surgical glioma resection in the Department of Neurosurgery, Soochow University-Affiliated First Hospital between September, 2006 and December, 2010. Levels of miR-29a and B7-H3 mRNAs in the tissue specimens and human glioma U87 cells were determined by Real-time PCR. U87 cells were transfected with miR-29a mimics, and B7-H3 expression and invasive capacity in the transfectants were assessed by flow cytometry and transwell migration assay, respectively. The expression of invasion-related chemical chemokines was analyzed by flow cytometry, and the ability of miR-29a to bind to CXCR4 was predicted by the miRtarbase software. Results: In glioma tissue specimens, miR-29a and B7-H3 mRNA levels were inversely correlated. In vitro, miR-29a down-regulated B7-H3 mRNAs expression in U87 cells and miR-29a-mediated down-regulation of B7-H3 resulted in a significant decrease in the invasive ability but not proliferative activity in U87 cells. Parallel to down-regulation of B7-H3 expression, CXCR4 expression was also down-regulated in U87 cells transfected with miR-29a mimics. No miR-29a binding sites were detected in the CXCR4 gene. Conclusions: In human glioma, miR-29a can effectively down-regulate B7-H3 expression and inhibit invasive activity. It is likely that these effects of miR-29a were at least attributable dwon-regulated expression of CXCR4.
2015, 22(1):34-40.
DOI: 10.3872/j.issn.1007-385X.2015.1.006
Abstract:
Objective: To develop a methodology of preparing novel C-phycocyanin-carboxymethyl chitosan nanoparticles(C-PC/CMCNPs) and determine the effect of C-PC/CMCNPs on the growth of HeLa cells. Methods: An orthogonal experiment was designed with the particle diameter and entrapment efficiency as index and CMC: C-PC mass ratio, CMC concentration, and CaCl2 concentration as factors to determine the best preparing condition of C-PC/CMCNPs. The effects of the generated C-PC/CMCNPs on the growth and apoptosis of HeLa cells were assessed by CCK-8 assay and flow cytometry respectively. Caspase-3 protein expression in HeLa cells was quantified by Western blotting. Results: The optimal condition for C-PC/CMCNPs preparations were as follows: C-PC to CMC ratio of 1∶2, CMC concentration of 1 mg/ml and CaCl2 concentration of 1 mg/ml. The C-PC/CMCNPs prepared in these optimal conditions had a high entrapment efficiency with an average particle diameter of 118.4 nm. C-PC, CMC and C-PC/CMCNPs were all capable of inhibiting proliferation and inducing apoptosis in HeLa cells by up-regulating the expression of the caspase-3 protein, but the effect of C-PC/CMCNPs was significantly more pronounced (P<0.05). Conclusions: We have optimized the conditions of preparing C-PC/CMCNPs. The nanoparticles prepared under these conditions have an acceptable safety profile of sustained C-PC release and possess a caspase-3-dependent anti-tumor activity, suggesting potential clinical implications.
Objective: To develop a methodology of preparing novel C-phycocyanin-carboxymethyl chitosan nanoparticles(C-PC/CMCNPs) and determine the effect of C-PC/CMCNPs on the growth of HeLa cells. Methods: An orthogonal experiment was designed with the particle diameter and entrapment efficiency as index and CMC: C-PC mass ratio, CMC concentration, and CaCl2 concentration as factors to determine the best preparing condition of C-PC/CMCNPs. The effects of the generated C-PC/CMCNPs on the growth and apoptosis of HeLa cells were assessed by CCK-8 assay and flow cytometry respectively. Caspase-3 protein expression in HeLa cells was quantified by Western blotting. Results: The optimal condition for C-PC/CMCNPs preparations were as follows: C-PC to CMC ratio of 1∶2, CMC concentration of 1 mg/ml and CaCl2 concentration of 1 mg/ml. The C-PC/CMCNPs prepared in these optimal conditions had a high entrapment efficiency with an average particle diameter of 118.4 nm. C-PC, CMC and C-PC/CMCNPs were all capable of inhibiting proliferation and inducing apoptosis in HeLa cells by up-regulating the expression of the caspase-3 protein, but the effect of C-PC/CMCNPs was significantly more pronounced (P<0.05). Conclusions: We have optimized the conditions of preparing C-PC/CMCNPs. The nanoparticles prepared under these conditions have an acceptable safety profile of sustained C-PC release and possess a caspase-3-dependent anti-tumor activity, suggesting potential clinical implications.
2015, 22(1):41-46.
DOI: 10.3872/j.issn.1007-385X.2015.1.007
Abstract:
Objective : To investigate the effect of arsenic trioxide (As2O3) on proliferation, cycle progression, apoptosis and radio-sensitivity of esophageal carcinoma cells under hypoxia. Methods: Human esophageal carcinoma Eca109 cells were treated with different concentrations of As2O3 or doses of radiation under a hypoxic condition mimicked cobalt chloride (CoCl2). At different time points after treatment, cell viability was determined by MTT assay, cell cycle progression and apoptosis by flow cytometry (FCM), and expression of HIF-1α and p27 at the protein level by Western blotting. Results: In time- and dose-dependent manners, As2O3 inhibited Eca109 cell proliferation similarly under both normoxic and hypoxic conditions (P>0.05). However, radiation-mediated inhibition of Eca109 cell proliferation was significantly less strong under hypoxia than under normoxia (P<0.05). Compared with normoxia, hypoxia increased cell cycle arrest at the G0/G1 phase and decreased the proportion of cells at the G2/M phase (P<0.05). As2O3 induced cell cycle arrest at the G2/M phase and reduced the proportion of cells at the G0/G1 phase under hypoxia. The combination of As2O3 and irradiation resulted in more significant apoptosis in Eca109 cells as compared with the use of As2O3 and irradiation each alone (P<0.05). Under hypoxia, HIF-1α and p27 protein contents were significantly increased as compared with normoxia (P<0.05), but the increase was significantly attenuated by As2O3 (P<0.05). Conclusions: Under hypoxia, As2O3 may increase the sensitivity esophageal carcinoma cells to radiation, possibly through down-regulating the expression of HIF-1α and its down-stream target p27, thus releasing G0/G1 phase arrest and inducing G2/M phase arrest.
Objective : To investigate the effect of arsenic trioxide (As2O3) on proliferation, cycle progression, apoptosis and radio-sensitivity of esophageal carcinoma cells under hypoxia. Methods: Human esophageal carcinoma Eca109 cells were treated with different concentrations of As2O3 or doses of radiation under a hypoxic condition mimicked cobalt chloride (CoCl2). At different time points after treatment, cell viability was determined by MTT assay, cell cycle progression and apoptosis by flow cytometry (FCM), and expression of HIF-1α and p27 at the protein level by Western blotting. Results: In time- and dose-dependent manners, As2O3 inhibited Eca109 cell proliferation similarly under both normoxic and hypoxic conditions (P>0.05). However, radiation-mediated inhibition of Eca109 cell proliferation was significantly less strong under hypoxia than under normoxia (P<0.05). Compared with normoxia, hypoxia increased cell cycle arrest at the G0/G1 phase and decreased the proportion of cells at the G2/M phase (P<0.05). As2O3 induced cell cycle arrest at the G2/M phase and reduced the proportion of cells at the G0/G1 phase under hypoxia. The combination of As2O3 and irradiation resulted in more significant apoptosis in Eca109 cells as compared with the use of As2O3 and irradiation each alone (P<0.05). Under hypoxia, HIF-1α and p27 protein contents were significantly increased as compared with normoxia (P<0.05), but the increase was significantly attenuated by As2O3 (P<0.05). Conclusions: Under hypoxia, As2O3 may increase the sensitivity esophageal carcinoma cells to radiation, possibly through down-regulating the expression of HIF-1α and its down-stream target p27, thus releasing G0/G1 phase arrest and inducing G2/M phase arrest.
2015, 22(1):47-51.
DOI: 10.3872/j.issn.1007-385X.2015.1.008
Abstract:
Objective: To investigate the effect of γ-secretase inhibitor (GSI) on proliferation and apoptosis of B lymphoma in vitro and in vivo. Methods: In experiments in vitro, Burkitt lymphoma (BL) Namalwa cells were treated with GSI at different concentrations. At different time points after treatment, cell viability and apoptosis were assessed by MTT assay and flow cytometry, respectively. Apoptosis-related proteins caspase-3 and caspase-9 and Notch signaling pathway proteins were assessed by Western blotting. In experiments in vivo, nude mice were injected with Namalwa cells. Mice that developed tumor lesions after xenograft implantation were treated with GSI or PBS. Tumor development was comparatively assessed in the two treatment groups. Results: GSI inhibited Namalwa cell proliferation in time- and dose-dependent manners; at 48 h and 72 h after treatment, the IC50 value was 2.14 μmol/L and 0.61 μmol/L, respectively. GSI also induced Namalwa cell apoptosis in a time-dependent manner; (17.71±1.87)% of Namalwa cells treated with GSI at 125 μmol/L but only 3.42±0.95 % of cells treated with PBS underwent apoptosis at 24 h (P<0.01), and these numbers (43.68±0.53)% and (4.65±0.8)%, respectively, at 48 (P<0.01). Contents of caspase-3 and caspase-9 and intracellular Notch proteins were decreased but activated caspase-9 protein content was increased in a time-dependent manner in Namalwa cells after GSI treatment. In tumor-bearing nude mice, GSI treatment for 13 days resulted in a significant decrease in the tumor volume as compared with the PBS control (2.199±0.183% vs 1.15±0.399%, P＜0.01). Correspondingly, a significantly higher rate of apoptosis was detected in tumors from GSI-treated animals (5.3±0.48)% than in PBS-treated animals (2.1±0.26) %(P<0.01). Conclusions: GSI can efficiently inhibit the proliferation and promote apoptosis of B lymphoma cells, at least through regulating caspase-3 and caspase-9 and Notch signaling pathway.
Objective: To investigate the effect of γ-secretase inhibitor (GSI) on proliferation and apoptosis of B lymphoma in vitro and in vivo. Methods: In experiments in vitro, Burkitt lymphoma (BL) Namalwa cells were treated with GSI at different concentrations. At different time points after treatment, cell viability and apoptosis were assessed by MTT assay and flow cytometry, respectively. Apoptosis-related proteins caspase-3 and caspase-9 and Notch signaling pathway proteins were assessed by Western blotting. In experiments in vivo, nude mice were injected with Namalwa cells. Mice that developed tumor lesions after xenograft implantation were treated with GSI or PBS. Tumor development was comparatively assessed in the two treatment groups. Results: GSI inhibited Namalwa cell proliferation in time- and dose-dependent manners; at 48 h and 72 h after treatment, the IC50 value was 2.14 μmol/L and 0.61 μmol/L, respectively. GSI also induced Namalwa cell apoptosis in a time-dependent manner; (17.71±1.87)% of Namalwa cells treated with GSI at 125 μmol/L but only 3.42±0.95 % of cells treated with PBS underwent apoptosis at 24 h (P<0.01), and these numbers (43.68±0.53)% and (4.65±0.8)%, respectively, at 48 (P<0.01). Contents of caspase-3 and caspase-9 and intracellular Notch proteins were decreased but activated caspase-9 protein content was increased in a time-dependent manner in Namalwa cells after GSI treatment. In tumor-bearing nude mice, GSI treatment for 13 days resulted in a significant decrease in the tumor volume as compared with the PBS control (2.199±0.183% vs 1.15±0.399%, P＜0.01). Correspondingly, a significantly higher rate of apoptosis was detected in tumors from GSI-treated animals (5.3±0.48)% than in PBS-treated animals (2.1±0.26) %(P<0.01). Conclusions: GSI can efficiently inhibit the proliferation and promote apoptosis of B lymphoma cells, at least through regulating caspase-3 and caspase-9 and Notch signaling pathway.
2015, 22(1):52-56.
DOI: 10.3872/j.issn.1007-385X.2015.1.009
Abstract:
Objective: To investigate the relationship between the expression of estrogen receptorβand endocrine resistance to tamoxifen (TAM) in breast cancer. Methods: Human breast cancer MCF-7 cells transfected with ERα or ER β constructs, M/HK (negative control), M/siα (ERαhigh/ERβlow), and M/siβ (ERαhigh/ERβlow), respectively, were used. The resistance of these transfected cells to TAM was assessed by MTT assay, mRNA levels of the major drug-resistance related genes (MDR1, TOPOⅡ, LRP, and GST- π) by RT-PCR, and levels of p-Akt, p-ERK and PI3K/Akt (major components of the signaling pathways involved in drug-resistance) by Western blotting. Results: Compared with MCF-7 expressing M/HK, cells expressing ER βshowed enhanced proliferation inhibition mediated by TAM in a dose-dependent manner (P<0.05) : (45.788±1.641)% vs (24.288±1.170)% at 1 μmol/L， (57.899±1.583)% vs (31.499±1.978)% at 5 μmol/L, and (59.853±1.648)% vs (38.039±1.482)% at 10 μmol/L, had significantly lower (P<0.05) mRNA levels of MDR1 (0.431±0.032 vs 0.932±0.083), TOPOⅡ (0.234±0.008 vs 0.391±0.002), and LRP (0.47±0.028 vs 0.586±0.036), and had significantly decreased (P<0.05) levels of p-Akt (0.224±0.006) vs (0.437±0.007) and p-ERK (0.367±0.015 vs 0.756±0.039). Conclusion: ER β may alter the resistance of human breast cancer cells to TAM, possibly through down-regulating the expression of drug-resistance genes and activating PI3K/AKT and ERK signal pathways.
Objective: To investigate the relationship between the expression of estrogen receptorβand endocrine resistance to tamoxifen (TAM) in breast cancer. Methods: Human breast cancer MCF-7 cells transfected with ERα or ER β constructs, M/HK (negative control), M/siα (ERαhigh/ERβlow), and M/siβ (ERαhigh/ERβlow), respectively, were used. The resistance of these transfected cells to TAM was assessed by MTT assay, mRNA levels of the major drug-resistance related genes (MDR1, TOPOⅡ, LRP, and GST- π) by RT-PCR, and levels of p-Akt, p-ERK and PI3K/Akt (major components of the signaling pathways involved in drug-resistance) by Western blotting. Results: Compared with MCF-7 expressing M/HK, cells expressing ER βshowed enhanced proliferation inhibition mediated by TAM in a dose-dependent manner (P<0.05) : (45.788±1.641)% vs (24.288±1.170)% at 1 μmol/L， (57.899±1.583)% vs (31.499±1.978)% at 5 μmol/L, and (59.853±1.648)% vs (38.039±1.482)% at 10 μmol/L, had significantly lower (P<0.05) mRNA levels of MDR1 (0.431±0.032 vs 0.932±0.083), TOPOⅡ (0.234±0.008 vs 0.391±0.002), and LRP (0.47±0.028 vs 0.586±0.036), and had significantly decreased (P<0.05) levels of p-Akt (0.224±0.006) vs (0.437±0.007) and p-ERK (0.367±0.015 vs 0.756±0.039). Conclusion: ER β may alter the resistance of human breast cancer cells to TAM, possibly through down-regulating the expression of drug-resistance genes and activating PI3K/AKT and ERK signal pathways.
2015, 22(1):57-61.
DOI: 10.3872/j.issn.1007-385X.2015.1.010
Abstract:
Objective: To optimize the preparation of polyamidoamine dendrimer-CXCR4-shRNA nanoparticles (PAMAM-shRNA) and study the inhibitory effect of PAMAM-shRNA nanoparticles on the proliferation of renal carcinoma cells in vitro. Methods: The seventh generation of polyamidoamine dendrimer (PAMAM-D) and CXCR4-shRNA were mixed at a mass ratio of 1∶0.73) at room temperature. The morphology and structure of PAMAM-shRNA by transmission electron microscopy, and determined the size of the nanoparticles were analyzed by laser particle size analyzer. Human renal carcinoma A498 cells were transfected with PAMAM-shRNA,CXCR4-shRNA and PAMAM-D respectively. After transfection, cell viability was assessed by MTT assays, cell apoptosis by flow cytometry, and CXCR4 mRNA abundance by Real-time PCR. Results: The prepared PAMAM-shRNA nanoparticles were evenly distributed and non-adherent with a mean diameter of 176.5±25.48 nm. PAMAM-shRNA effectively inhibited A498 proliferation in time- and dose-dependent manners, and the highest proliferation inhibition rate was up to （66.5±2.7)%. PAMAM-shRNA induced A498 cell apoptosis. CXCR4 mRNA abundance was significantly decreased (P<0.05) in cells transfected with PAMAM-shRNA (0.29±0035) than in cells transfected with CXCR4-shRNA (0.70±0.084). Conclusions: PAMAM dendrimers may efficiently mediate the entry of CXCR4-shRNA into renal carcinoma cells, where they exert proliferation-inhibitory and apoptosis-promoting activities. These observations suggest that PAMAM-shRNA nanoparticles may have a potential clinical application in gene therapy of renal cancer.
Objective: To optimize the preparation of polyamidoamine dendrimer-CXCR4-shRNA nanoparticles (PAMAM-shRNA) and study the inhibitory effect of PAMAM-shRNA nanoparticles on the proliferation of renal carcinoma cells in vitro. Methods: The seventh generation of polyamidoamine dendrimer (PAMAM-D) and CXCR4-shRNA were mixed at a mass ratio of 1∶0.73) at room temperature. The morphology and structure of PAMAM-shRNA by transmission electron microscopy, and determined the size of the nanoparticles were analyzed by laser particle size analyzer. Human renal carcinoma A498 cells were transfected with PAMAM-shRNA,CXCR4-shRNA and PAMAM-D respectively. After transfection, cell viability was assessed by MTT assays, cell apoptosis by flow cytometry, and CXCR4 mRNA abundance by Real-time PCR. Results: The prepared PAMAM-shRNA nanoparticles were evenly distributed and non-adherent with a mean diameter of 176.5±25.48 nm. PAMAM-shRNA effectively inhibited A498 proliferation in time- and dose-dependent manners, and the highest proliferation inhibition rate was up to （66.5±2.7)%. PAMAM-shRNA induced A498 cell apoptosis. CXCR4 mRNA abundance was significantly decreased (P<0.05) in cells transfected with PAMAM-shRNA (0.29±0035) than in cells transfected with CXCR4-shRNA (0.70±0.084). Conclusions: PAMAM dendrimers may efficiently mediate the entry of CXCR4-shRNA into renal carcinoma cells, where they exert proliferation-inhibitory and apoptosis-promoting activities. These observations suggest that PAMAM-shRNA nanoparticles may have a potential clinical application in gene therapy of renal cancer.
2015, 22(1):62-66.
DOI: 10.3872/j.issn.1007-385X.2015.1.011
Abstract:
Objective : To analyze the correlation between the expression of phosphoinositide-3 kinase catalytic subunit delta (PIK3CD) and the clinicopathologic characteristics in gastric cancer tissues. Methods: A total of 86 paired tumorous and normal gastric tissue specimens were collected from gastric cancer patients who underwent surgical resection in the Department of Gastrointestinal Surgery, Liaoning Medical University-Affiliated First Hospital in Jinzhou between May 2005 and December 2007. The expression of PIK3CD at the protein level in these specimens was assessed by immunohistochemistry. The possible correlation between PIK3CD expression and the patients'clinicopathological characteristics and prognosis was analyzed by χ2 and Spearman correlation test. The influence of PIK3CD on 5-year survival and survival length was analyzed by Kaplan-Meier test. Results: The immunoreactive PIK3CD protein signal in the tumor proper was significantly stronger in the tumor proper than in the matched adjacent tissue (P=0.006). The intensity of PIK3CD protein signal was positively correlated with infiltration (P=0.005), but not with sex, age, differention, clinical stage, lympha nodes metastasis and distance metastasis (P>0.05). The 5-year survival rate was 18.9% and 60.6% in patients with high and low levels of PIK3CD respectively (P<0.05). The median survival length was 31.0 (95%CI: 23.9-38.1) months in patients with high levels of PIK3CD and 60.6 (95%CI: 53.8-67.4) months in patients with low levels of PIK3CD (χ2=19.791, P=0000). Elevated PIK3CD expression was an independent negative prognostic factor of gastric cancer (P= 0.000). Conclusion: PIK3CD may function as a pro-oncogeneand an independent prognostic factor in patients with gastric cancer.
Objective : To analyze the correlation between the expression of phosphoinositide-3 kinase catalytic subunit delta (PIK3CD) and the clinicopathologic characteristics in gastric cancer tissues. Methods: A total of 86 paired tumorous and normal gastric tissue specimens were collected from gastric cancer patients who underwent surgical resection in the Department of Gastrointestinal Surgery, Liaoning Medical University-Affiliated First Hospital in Jinzhou between May 2005 and December 2007. The expression of PIK3CD at the protein level in these specimens was assessed by immunohistochemistry. The possible correlation between PIK3CD expression and the patients'clinicopathological characteristics and prognosis was analyzed by χ2 and Spearman correlation test. The influence of PIK3CD on 5-year survival and survival length was analyzed by Kaplan-Meier test. Results: The immunoreactive PIK3CD protein signal in the tumor proper was significantly stronger in the tumor proper than in the matched adjacent tissue (P=0.006). The intensity of PIK3CD protein signal was positively correlated with infiltration (P=0.005), but not with sex, age, differention, clinical stage, lympha nodes metastasis and distance metastasis (P>0.05). The 5-year survival rate was 18.9% and 60.6% in patients with high and low levels of PIK3CD respectively (P<0.05). The median survival length was 31.0 (95%CI: 23.9-38.1) months in patients with high levels of PIK3CD and 60.6 (95%CI: 53.8-67.4) months in patients with low levels of PIK3CD (χ2=19.791, P=0000). Elevated PIK3CD expression was an independent negative prognostic factor of gastric cancer (P= 0.000). Conclusion: PIK3CD may function as a pro-oncogeneand an independent prognostic factor in patients with gastric cancer.
2015, 22(1):67-72.
DOI: 10.3872/j.issn.1007-385X.2015.1.012
Abstract:
Objective: To evaluate the effect of Helicobacter pylori (Hp) infection on the outcome of maintenance therapy with autologous dendritic cells-cytokine induced killer (DC-CIK) cells in gastric cancer patients. Methods: Seventy-two patients with advanced gastric cancer, aged 29-90 years, who were admitted to the Department of Oncology of No. 174 Hospital of PLA between June, 2010 and June, 2012 were included in this study. Based on the gastroscopic findings, patients were divided into two groups: Hp-positive and Hp-negative. Both groups received two courses of autologous DC-CIK immunotherapy after surgery or radiotherapy and chemotherapy. At the end of treatment, DC differentiation and maturation, treatment efficacy and patients outcomes in the two groups were analyzed. Results: No significant difference was observed in the morphology and maturation status of DCs between two groups (P>0.05). Levels of CD83 and CD86 on the surface of DCs were significantly higher in the Hp-positive group than those in the Hp-negative group (P<0.01), but surface levels of HLA-DR in DCs had no significant difference between two groups (P>0.05). In the Hp-positive group, the quality of life KPS score and levels of T lymphocyte markers (CD3+, CD4+ and CD8+) were significantly increased (P<0.05) but levels of tumor markers CEA, CA199 and CA724 were significantly decreased (P<0.05) after treatment. In the Hp-negative group, levels of tumor markers and the T lymphocyte markers were significantly different before and after treatment (P<0.05), but the KPS score was not significantly improved (P>0.05). Hp-positive patients had better KPS score, higher levels of CEA and CA199, and more abundant CD3+ CD4+T cells than Hp-negative patients (P<0.05）. In contrast, the two groups did not significantly differ in tumor stability, CA724 expression, and the number of CD8+T cells (P>0.05）. At the end of 2-years follow-up, the median survival time was 12.64 months in the HP-positive group and 11.42 months in the HP-negative group (P<0.05）. Conclusion: Hp infection may help stabilize the tumor size, improve the quality of life, and prolong the survival time in patients with advanced gastric cancer undergoing maintenance therapy with DC-CIK cells.
Objective: To evaluate the effect of Helicobacter pylori (Hp) infection on the outcome of maintenance therapy with autologous dendritic cells-cytokine induced killer (DC-CIK) cells in gastric cancer patients. Methods: Seventy-two patients with advanced gastric cancer, aged 29-90 years, who were admitted to the Department of Oncology of No. 174 Hospital of PLA between June, 2010 and June, 2012 were included in this study. Based on the gastroscopic findings, patients were divided into two groups: Hp-positive and Hp-negative. Both groups received two courses of autologous DC-CIK immunotherapy after surgery or radiotherapy and chemotherapy. At the end of treatment, DC differentiation and maturation, treatment efficacy and patients outcomes in the two groups were analyzed. Results: No significant difference was observed in the morphology and maturation status of DCs between two groups (P>0.05). Levels of CD83 and CD86 on the surface of DCs were significantly higher in the Hp-positive group than those in the Hp-negative group (P<0.01), but surface levels of HLA-DR in DCs had no significant difference between two groups (P>0.05). In the Hp-positive group, the quality of life KPS score and levels of T lymphocyte markers (CD3+, CD4+ and CD8+) were significantly increased (P<0.05) but levels of tumor markers CEA, CA199 and CA724 were significantly decreased (P<0.05) after treatment. In the Hp-negative group, levels of tumor markers and the T lymphocyte markers were significantly different before and after treatment (P<0.05), but the KPS score was not significantly improved (P>0.05). Hp-positive patients had better KPS score, higher levels of CEA and CA199, and more abundant CD3+ CD4+T cells than Hp-negative patients (P<0.05）. In contrast, the two groups did not significantly differ in tumor stability, CA724 expression, and the number of CD8+T cells (P>0.05）. At the end of 2-years follow-up, the median survival time was 12.64 months in the HP-positive group and 11.42 months in the HP-negative group (P<0.05）. Conclusion: Hp infection may help stabilize the tumor size, improve the quality of life, and prolong the survival time in patients with advanced gastric cancer undergoing maintenance therapy with DC-CIK cells.
2015, 22(1):73-78.
DOI: 10.3872/j.issn.1007-385X.2015.1.013
Abstract:
Objective: To study the expression and putative role of a novel transient receptor potential calcium channel subfamily V member 6 (TRPV6) in stomach cancer. Methods： Sixty-five pairs of tumor and surrounding tissue specimens were collected from patients with pathologically confirmed stomach cancer who underwent surgical resection in our hospital between May, 2010 and March, 2013. TRPV6 protein in these specimens was assessed by immunohistochemical SP staining. To evaluate the putative functional role for TRPV6 in stomach cancer, human stomach cancer MGC-803 cells were treated with 2-amino ethyl two phenyl boronic acid (2-APB), a TRPV6 channel blocker. After treatment, cell proliferation, apoptosis and migration were assessed by CCK-8 assay, flow cytometry and transwell assay, respectively, and TRPV6, AKT/p-AKT, p-GSK3 protein contents were analyzed by Western blotting. Results: TRPV6 protein was detected in 95.4% (62/65) of cancerous tissue specimens but only in 36.9% (24/65) of the corresponding non-cancerous tissue specimens (P<0.01). Immunoreactive TRPV6 signal was positively associated with the tumor size, lymph, distant metastasis and Dukes stage (P<0.05). The TRPV6 blocker 2-APB significantly inhibited MGC-803 cell proliferation, induced MGC-803 cell apoptosis and inhibited MGC-803 cell migration (P<0.05). Moreover, 2-APB treatment resulted in significant decreases in TRPV6, p-AKT, and p-GSK3β proteins in a dose-dependent manner in MGC-803 cells (P<005). Conclusions: TRPV6 is highly expressed in stomach cancer, where it promotes cancer cell proliferation and inhibits cancer cell apoptosis. Down-regulation of p-AKT, p-GSK-3 β protein expressions may be possible mechanisms underlying the tumorigenic activity of TRPV6.
Objective: To study the expression and putative role of a novel transient receptor potential calcium channel subfamily V member 6 (TRPV6) in stomach cancer. Methods： Sixty-five pairs of tumor and surrounding tissue specimens were collected from patients with pathologically confirmed stomach cancer who underwent surgical resection in our hospital between May, 2010 and March, 2013. TRPV6 protein in these specimens was assessed by immunohistochemical SP staining. To evaluate the putative functional role for TRPV6 in stomach cancer, human stomach cancer MGC-803 cells were treated with 2-amino ethyl two phenyl boronic acid (2-APB), a TRPV6 channel blocker. After treatment, cell proliferation, apoptosis and migration were assessed by CCK-8 assay, flow cytometry and transwell assay, respectively, and TRPV6, AKT/p-AKT, p-GSK3 protein contents were analyzed by Western blotting. Results: TRPV6 protein was detected in 95.4% (62/65) of cancerous tissue specimens but only in 36.9% (24/65) of the corresponding non-cancerous tissue specimens (P<0.01). Immunoreactive TRPV6 signal was positively associated with the tumor size, lymph, distant metastasis and Dukes stage (P<0.05). The TRPV6 blocker 2-APB significantly inhibited MGC-803 cell proliferation, induced MGC-803 cell apoptosis and inhibited MGC-803 cell migration (P<0.05). Moreover, 2-APB treatment resulted in significant decreases in TRPV6, p-AKT, and p-GSK3β proteins in a dose-dependent manner in MGC-803 cells (P<005). Conclusions: TRPV6 is highly expressed in stomach cancer, where it promotes cancer cell proliferation and inhibits cancer cell apoptosis. Down-regulation of p-AKT, p-GSK-3 β protein expressions may be possible mechanisms underlying the tumorigenic activity of TRPV6.
2015, 22(1):79-83.
DOI: 10.3872/j.issn.1007-385X.2015.1.014
Abstract:
Objective: To assess changes in the proportion of CD4 + CD25 + FoxP3 + regulatory T (Treg) cells and their cytokine production in the peripheral blood of gastric cancer patients after immunotherapy with DC-CIK cells. Methods: This study enrolled 158 patients with gastric cancer who were admitted to the Biotherapy Center, Qingdao Center Hospital between April, 2009 and December, 2013. They received immunotherapy with DC-CIK cells for one to four cycles. At the end of the treatment, peripheral blood was collected. The proportion of Treg cells and levels of Treg cell-derived cytokines IL-10 and TGF-β1 were assessed by flow cytometry and ELISA respectively. Results: In patients (n=75) who received more than 3 cycles of immunotherapy, the proportion of Treg cells and levels of IL-10 and TGF-β1 were decreased significantly after DC-CIK therapy (Z＝-2.65, -3.21, -3.95, P<0.05). In patients who received less than 2 cycles of immunotherapy, the proportion of Treg cells and levels of IL-10 and TGF-β1 did not change significantly after treatment (P>0.05). In patients receiving the same number of cycle of treatment, the proportion of Treg cells and levels of IL-10 and TGF-β1 were not significantly different between recurrent and non-recurrence cases (P>0.05). Conclusion: Immunotherapy with DC-CIK cells for ≥3 cycles may significantly decrease the proportion of Treg cells and their cytokine levels in the peripheral blood. In the patients receiving treatment for the same number of cycles, recurrence has not effect on the proportion of Treg cells.
Objective: To assess changes in the proportion of CD4 + CD25 + FoxP3 + regulatory T (Treg) cells and their cytokine production in the peripheral blood of gastric cancer patients after immunotherapy with DC-CIK cells. Methods: This study enrolled 158 patients with gastric cancer who were admitted to the Biotherapy Center, Qingdao Center Hospital between April, 2009 and December, 2013. They received immunotherapy with DC-CIK cells for one to four cycles. At the end of the treatment, peripheral blood was collected. The proportion of Treg cells and levels of Treg cell-derived cytokines IL-10 and TGF-β1 were assessed by flow cytometry and ELISA respectively. Results: In patients (n=75) who received more than 3 cycles of immunotherapy, the proportion of Treg cells and levels of IL-10 and TGF-β1 were decreased significantly after DC-CIK therapy (Z＝-2.65, -3.21, -3.95, P<0.05). In patients who received less than 2 cycles of immunotherapy, the proportion of Treg cells and levels of IL-10 and TGF-β1 did not change significantly after treatment (P>0.05). In patients receiving the same number of cycle of treatment, the proportion of Treg cells and levels of IL-10 and TGF-β1 were not significantly different between recurrent and non-recurrence cases (P>0.05). Conclusion: Immunotherapy with DC-CIK cells for ≥3 cycles may significantly decrease the proportion of Treg cells and their cytokine levels in the peripheral blood. In the patients receiving treatment for the same number of cycles, recurrence has not effect on the proportion of Treg cells.
2015, 22(1):84-88.
DOI: 10.3872/j.issn.1007-385X.2015.1.015
Abstract:
Objective: To evaluate the effectiveness and safety of autologous cytokine-induced killer cells in the treatment of advanced non-small cell lung cancer (NSCLC) after radiotherapy and chemotherapy. Methods: A total of 72 patients with advanced NSCLC who were admitted to our department between June, 2012 and March, 2014 and had undergone radiotherapy and chemotherapy were enrolled. One month after the last radiotherapy and chemotherapy, 38 patients were treated with autologous cytokine-induced killer (CIK) cells for two cycles (5 days each) with an interval of 2 months in addition to the optimal maintenance treatment regimen and the remaining 34 patients were given maintenance treatment only as controls. One week before and one week after treatment with CIK cells, proportions of CD3 +, CD4 +, CD8 +, CD4 +/CD8 +, and CD4 +CD25 + regulatory T (Treg) cells in the peripheral blood were determined by flow cytometry, levels of IL-2, IL-12 and IFN-γ were measured with ELISA, and the Karnofsky performance score (KPS) was calculated. Results: The proportion of CD4 + T cells and the CD4/CD8 + T cell ratio were significantly higher (P<0.05), the proportion of CD4 +CD25 + Treg cells was significantly lower (P<0.05), the levels of IL-12, IFN-γ were significantly increased (P<0.01), and the KPS score was significantly higher (P<0.05) in patients receiving autologous CIKs as compared with control subjects. None of the patients suffered from transient fever or chills in the process of CIK transfusion, and no other side effects were observed. Conclusion: Autologous CIKs may improve the immune function and the quality of life in patients with advanced NSCLC, thus offering an effective and safe treatment option for NSCLC.
Objective: To evaluate the effectiveness and safety of autologous cytokine-induced killer cells in the treatment of advanced non-small cell lung cancer (NSCLC) after radiotherapy and chemotherapy. Methods: A total of 72 patients with advanced NSCLC who were admitted to our department between June, 2012 and March, 2014 and had undergone radiotherapy and chemotherapy were enrolled. One month after the last radiotherapy and chemotherapy, 38 patients were treated with autologous cytokine-induced killer (CIK) cells for two cycles (5 days each) with an interval of 2 months in addition to the optimal maintenance treatment regimen and the remaining 34 patients were given maintenance treatment only as controls. One week before and one week after treatment with CIK cells, proportions of CD3 +, CD4 +, CD8 +, CD4 +/CD8 +, and CD4 +CD25 + regulatory T (Treg) cells in the peripheral blood were determined by flow cytometry, levels of IL-2, IL-12 and IFN-γ were measured with ELISA, and the Karnofsky performance score (KPS) was calculated. Results: The proportion of CD4 + T cells and the CD4/CD8 + T cell ratio were significantly higher (P<0.05), the proportion of CD4 +CD25 + Treg cells was significantly lower (P<0.05), the levels of IL-12, IFN-γ were significantly increased (P<0.01), and the KPS score was significantly higher (P<0.05) in patients receiving autologous CIKs as compared with control subjects. None of the patients suffered from transient fever or chills in the process of CIK transfusion, and no other side effects were observed. Conclusion: Autologous CIKs may improve the immune function and the quality of life in patients with advanced NSCLC, thus offering an effective and safe treatment option for NSCLC.
2015, 22(1):89-95.
DOI: 10.3872/j.issn.1007-385X.2015.1.016
Abstract:
Objective: This study aimed to evaluate the difference between the first/second stimulation signal-based T lymphocyte amplification technology (PensonGenTM) and cytokine-induced killer (CIK) technology in the expansion and differentiation of T cells in vitro. Methods: Peripheral blood mononuclear cells (PBMCs), isolated from 8 healthy volunteers, were expanded using the CIK technology (CIK group), PersonGen TM human T lymphocyte activation technology (PersonGen TM group), and combined techniques of PensonGen TM and CIK method (C+P group), respectively. The proliferative activity and T cell subsets in different groups of cells were assessed by hemocytometer counting and flow cytometry analysis. The cytotoxic activities of the expanded T cells were examined in vitro using myeloma RPMI 8226 and leukemia K562 cells as targets. Results: After three weeks of culture, cells in all three groups (CIK, PersonGen TM , and C+P group) were successfully expanded, the multiples were 254±56, 863±218, and 793.3±395, respectively, and the proportion of CD3 + cells were （51.61±14.95）%, （99.21±0.79）%, and （96.14±5.16）%, respectively. At the end of 3-week culture, CD3 +CD4 + cells were expanded approximately five times in CIK group and160 times in both C+P and PersonGen TM group groups, CD3 +CD8 + cells were expanded nearly 500 times in CIK group and up to 1 000 times in both PersonGen TM and C+P group groups, and CD3 +CD56 + NKT cells were expanded approximately 700 times and close to 1 000 times in the other two groups. At an effector-to-target cell ratio of 5〖DK〗∶1, the cytotoxic activity of the cells expanded by CIK technology, PersonGen TM technology and CIK plus PersonGen TM technologies were （45.53±19.16）%, （36.53±10.6）% and （53.17±5.8)%, respectively, when K562 cells were used as a target, and were （41.23±12.5）%, （38.83±4.3）% and （45.73±7.48）%, respectively, when RPMI 8226 cells were used as a target. The expanded T cells in PersonGen TM group had slight higher cytotoxicity against both K562 and RPMI 8226 cells than cells expanded in CIK group (P>0.05). Conclusion: Compared to CIK technology, secondary stimulation signal-based PersonGen TM technology is more efficient in the expansion of CD3 +CD8 + cell population from PBMCs, resulting in an enhanced anti-tumor activity.
Objective: This study aimed to evaluate the difference between the first/second stimulation signal-based T lymphocyte amplification technology (PensonGenTM) and cytokine-induced killer (CIK) technology in the expansion and differentiation of T cells in vitro. Methods: Peripheral blood mononuclear cells (PBMCs), isolated from 8 healthy volunteers, were expanded using the CIK technology (CIK group), PersonGen TM human T lymphocyte activation technology (PersonGen TM group), and combined techniques of PensonGen TM and CIK method (C+P group), respectively. The proliferative activity and T cell subsets in different groups of cells were assessed by hemocytometer counting and flow cytometry analysis. The cytotoxic activities of the expanded T cells were examined in vitro using myeloma RPMI 8226 and leukemia K562 cells as targets. Results: After three weeks of culture, cells in all three groups (CIK, PersonGen TM , and C+P group) were successfully expanded, the multiples were 254±56, 863±218, and 793.3±395, respectively, and the proportion of CD3 + cells were （51.61±14.95）%, （99.21±0.79）%, and （96.14±5.16）%, respectively. At the end of 3-week culture, CD3 +CD4 + cells were expanded approximately five times in CIK group and160 times in both C+P and PersonGen TM group groups, CD3 +CD8 + cells were expanded nearly 500 times in CIK group and up to 1 000 times in both PersonGen TM and C+P group groups, and CD3 +CD56 + NKT cells were expanded approximately 700 times and close to 1 000 times in the other two groups. At an effector-to-target cell ratio of 5〖DK〗∶1, the cytotoxic activity of the cells expanded by CIK technology, PersonGen TM technology and CIK plus PersonGen TM technologies were （45.53±19.16）%, （36.53±10.6）% and （53.17±5.8)%, respectively, when K562 cells were used as a target, and were （41.23±12.5）%, （38.83±4.3）% and （45.73±7.48）%, respectively, when RPMI 8226 cells were used as a target. The expanded T cells in PersonGen TM group had slight higher cytotoxicity against both K562 and RPMI 8226 cells than cells expanded in CIK group (P>0.05). Conclusion: Compared to CIK technology, secondary stimulation signal-based PersonGen TM technology is more efficient in the expansion of CD3 +CD8 + cell population from PBMCs, resulting in an enhanced anti-tumor activity.
2015, 22(1):96-98.
DOI: 10.3872/j.issn.1007-385X.2015.1.017
Abstract:
目的：探讨原发性肝细胞癌（hepatocellular carcinoma，HCC）患者外周血单个核细胞（peripheral blood mononrclear cells，PBMCs）IL-22和IL-17 mRNA的表达水平及其临床意义。方法：收集2011年4月至10月暨南大学医学院附属深圳市人民医院感染内科经临床确诊为HCC患者40例，所有患者均为乙型肝炎表面抗原阳性。同时选取来自本科室健康人的外周静脉血20例作为对照。以常规淋巴细胞分层液密度梯度离心法分离得到PBMCs，采用RT- PCR法检测HCC患者组和健康人对照组PBMCs IL-22和IL-17 mRNA的表达水平。结果：与健康人对照组相比，HCC患者组IL-22 mRNA表达水平明显下降（0.309±0.044 vs 0.560±0.029，P＜0.01），而IL-17 mRNA的表达水平则显著升高（0.682±0.048 vs 0.541±0.038， P＜0.01）。结论： IL-17 mRNA的高表达可能参与HCC的发生、发展，而IL-22 mRNA低表达可能与HCC患者的近期预后不佳有关。
目的：探讨原发性肝细胞癌（hepatocellular carcinoma，HCC）患者外周血单个核细胞（peripheral blood mononrclear cells，PBMCs）IL-22和IL-17 mRNA的表达水平及其临床意义。方法：收集2011年4月至10月暨南大学医学院附属深圳市人民医院感染内科经临床确诊为HCC患者40例，所有患者均为乙型肝炎表面抗原阳性。同时选取来自本科室健康人的外周静脉血20例作为对照。以常规淋巴细胞分层液密度梯度离心法分离得到PBMCs，采用RT- PCR法检测HCC患者组和健康人对照组PBMCs IL-22和IL-17 mRNA的表达水平。结果：与健康人对照组相比，HCC患者组IL-22 mRNA表达水平明显下降（0.309±0.044 vs 0.560±0.029，P＜0.01），而IL-17 mRNA的表达水平则显著升高（0.682±0.048 vs 0.541±0.038， P＜0.01）。结论： IL-17 mRNA的高表达可能参与HCC的发生、发展，而IL-22 mRNA低表达可能与HCC患者的近期预后不佳有关。
2015, 22(1):99-103.
DOI: 10.3872/j.issn.1007-385X.2015.1.018
Abstract:
CD19是正常和恶性B淋巴细胞特异性表面蛋白，在B细胞的发育、增殖和分化以及恶性转化中发挥重要作用。因CD19在B淋巴细胞表达的特异性和恶性肿瘤表达的广泛性，使其成为一个颇具潜力的B淋巴细胞恶性肿瘤免疫治疗的分子靶点。靶向CD19的各种免疫治疗策略正在实验室和临床展开，包括Fc段修饰抗体、抗体-药物偶联物、双特异性抗体、嵌合抗原受体修饰T细胞等，在白血病和淋巴瘤中取得了令人鼓舞的治疗效果，有力地推动了靶向免疫治疗的发展。本文就近年靶向CD19治疗B细胞恶性肿瘤的研究进展加以综述。
CD19是正常和恶性B淋巴细胞特异性表面蛋白，在B细胞的发育、增殖和分化以及恶性转化中发挥重要作用。因CD19在B淋巴细胞表达的特异性和恶性肿瘤表达的广泛性，使其成为一个颇具潜力的B淋巴细胞恶性肿瘤免疫治疗的分子靶点。靶向CD19的各种免疫治疗策略正在实验室和临床展开，包括Fc段修饰抗体、抗体-药物偶联物、双特异性抗体、嵌合抗原受体修饰T细胞等，在白血病和淋巴瘤中取得了令人鼓舞的治疗效果，有力地推动了靶向免疫治疗的发展。本文就近年靶向CD19治疗B细胞恶性肿瘤的研究进展加以综述。
2015, 22(1):104-111.
DOI: 10.3872/j.issn.1007-385X.2015.1.019
Abstract:
NY-ESO-1是癌-睾丸抗原家族中的重要一员，在多种肿瘤组织中广泛表达，但在正常组织中几乎不表达。目前已经发现了20多个抗原表位,并在多种肿瘤患者体内检测到自发性免疫反应，包括体液和细胞免疫反应，NY-ESO-1的表达及体内自发性免疫应答被证实与肿瘤的诊断和预后有重要关联。NY-ESO-1被认为是肿瘤免疫治疗的理想靶抗原，以NY-ESO-1抗原为靶点的肿瘤治疗性疫苗、TCR基因修饰的T细胞、CAR修饰的T细胞等临床研究相继开展，展示了良好的前景。
NY-ESO-1是癌-睾丸抗原家族中的重要一员，在多种肿瘤组织中广泛表达，但在正常组织中几乎不表达。目前已经发现了20多个抗原表位,并在多种肿瘤患者体内检测到自发性免疫反应，包括体液和细胞免疫反应，NY-ESO-1的表达及体内自发性免疫应答被证实与肿瘤的诊断和预后有重要关联。NY-ESO-1被认为是肿瘤免疫治疗的理想靶抗原，以NY-ESO-1抗原为靶点的肿瘤治疗性疫苗、TCR基因修饰的T细胞、CAR修饰的T细胞等临床研究相继开展，展示了良好的前景。
2015, 22(1):112-115.
DOI: 10.3872/j.issn.1007-385X.2015.1.020
Abstract:
Hedgehog（HH）通路不仅在胰腺发育过程中起重要作用，而且在胰腺导管细胞癌（pancreatic ductal adenocarcinoma，PDAC）发生、发展过程中也出现异常活化。其转录因子Glis蛋白家族的活化，尤其是Gli1的活化可启动下游基因表达，主要起到促进癌细胞增殖、迁移、侵袭等作用。HH通路可以和包括核因子-κB（nuclear factor-κB，NF-κB）、K-RAS、表皮生长因子受体（epidermal growth factor receptor， EGFR）、Wnt通路、转化生长因子 β（transforming growth factor β，TGF-β）、丝裂原活化蛋白激酶3K10（mitogen-activated protein kinase kinase kinase 10，MAP3K10）和P53等在内的信号通路因子，通过不尽相同的机制相互作用，共同促进胰腺癌的发生和发展。本文就近年来对胰腺癌中的Hedgehog通路及其与其他通路相互作用的研究进展进行综述。
Hedgehog（HH）通路不仅在胰腺发育过程中起重要作用，而且在胰腺导管细胞癌（pancreatic ductal adenocarcinoma，PDAC）发生、发展过程中也出现异常活化。其转录因子Glis蛋白家族的活化，尤其是Gli1的活化可启动下游基因表达，主要起到促进癌细胞增殖、迁移、侵袭等作用。HH通路可以和包括核因子-κB（nuclear factor-κB，NF-κB）、K-RAS、表皮生长因子受体（epidermal growth factor receptor， EGFR）、Wnt通路、转化生长因子 β（transforming growth factor β，TGF-β）、丝裂原活化蛋白激酶3K10（mitogen-activated protein kinase kinase kinase 10，MAP3K10）和P53等在内的信号通路因子，通过不尽相同的机制相互作用，共同促进胰腺癌的发生和发展。本文就近年来对胰腺癌中的Hedgehog通路及其与其他通路相互作用的研究进展进行综述。
2015, 22(1):116-119.
DOI: 10.3872/j.issn.1007-385X.2015.1.021
Abstract:
端粒是真核生物染色体末端特异的核苷酸结构，主要由重复的“TTAGGG”序列组成并随着细胞分裂逐渐缩短。端粒酶能够防止端粒缩短、丢失、重排或者终端与终端的融合。人端粒酶逆转录酶（human telomerase reverse transcriptase，hTERT）现已被认为是端粒酶活性调控的重要组成部分。研究表明，过表达hTERT基因能够增强T细胞增殖能力，转入hTERT基因的T细胞增殖能力大大增强，这将成为肿瘤过继免疫治疗良好的细胞资源，从而为肿瘤生物治疗提供一种新的治疗方法。本文中，主要讨论T细胞端粒功能和端粒酶活性、端粒酶活性调控、过表达hTERT基因的T细胞增殖特征及转导hTERT基因的T细胞在肿瘤过继免疫治疗中的应用。
端粒是真核生物染色体末端特异的核苷酸结构，主要由重复的“TTAGGG”序列组成并随着细胞分裂逐渐缩短。端粒酶能够防止端粒缩短、丢失、重排或者终端与终端的融合。人端粒酶逆转录酶（human telomerase reverse transcriptase，hTERT）现已被认为是端粒酶活性调控的重要组成部分。研究表明，过表达hTERT基因能够增强T细胞增殖能力，转入hTERT基因的T细胞增殖能力大大增强，这将成为肿瘤过继免疫治疗良好的细胞资源，从而为肿瘤生物治疗提供一种新的治疗方法。本文中，主要讨论T细胞端粒功能和端粒酶活性、端粒酶活性调控、过表达hTERT基因的T细胞增殖特征及转导hTERT基因的T细胞在肿瘤过继免疫治疗中的应用。
2015, 22(1):120-124.
DOI: 10.3872/j.issn.1007-385X.2015.1.022
Abstract:
microRNA（miRNA）是内源性非编码单链小RNA，参与细胞增殖、凋亡与分化等多种重要生命活动的调控。近年来研究发现，miRNAs参与多种恶性肿瘤的演进，起抑癌基因或原癌基因的作用。miR-200c是上皮-间质转化（epithelial-mesenchymal, EMT)过程中的重要调节基因，除了在正常细胞的表型转换中起作用，还在多种类型癌细胞的表型转换中起调节作用，能促进或者抑制肿瘤的侵袭转移能力。此外，关于miR-200c的研究还涉及肿瘤的耐药性和凋亡抗性。研究证实，miR-200c能够促进或抑制肿瘤的侵袭转移，逆转肿瘤细胞的耐药性和凋亡抗性。本文主要对不同肿瘤中miR-200c对肿瘤进展的影响进行综述。
microRNA（miRNA）是内源性非编码单链小RNA，参与细胞增殖、凋亡与分化等多种重要生命活动的调控。近年来研究发现，miRNAs参与多种恶性肿瘤的演进，起抑癌基因或原癌基因的作用。miR-200c是上皮-间质转化（epithelial-mesenchymal, EMT)过程中的重要调节基因，除了在正常细胞的表型转换中起作用，还在多种类型癌细胞的表型转换中起调节作用，能促进或者抑制肿瘤的侵袭转移能力。此外，关于miR-200c的研究还涉及肿瘤的耐药性和凋亡抗性。研究证实，miR-200c能够促进或抑制肿瘤的侵袭转移，逆转肿瘤细胞的耐药性和凋亡抗性。本文主要对不同肿瘤中miR-200c对肿瘤进展的影响进行综述。
2015, 22(1):125-127.
DOI: 10.3872/j.issn.1007-385X.2015.1.023
Abstract:
联系方式
- 《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
- 邮编：200433
- 电话：021-81871002-22
- 电子邮箱：cjcb@biother.cn
- 网址:http://www.biother.cn
- 刊号：ISSN 1007-385X
- CN 31-1725/R
- 国内定价： ￥20元/册
您是第位访问者
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.
Chinese Journal Of Cancer Biotheray ® 2025 All Rights Reserved
Supported by:Beijing E-Tiller Technology Development Co., Ltd.