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Volume 18,Issue 1,2011 Table of Contents
2011, 18(1):1-6.
DOI: 10.3872/j.issn.1007-385X.2011.1.001
Abstract:
Adoptive cell therapy (ACT) of cancer has long been proposed, but was gradually forgotten due to its disappointing efficacy. In recent years, great improvement has been made in cancer ACT．ACT with adoptive T cells for immunotherapy of melanoma has achieved impressive outcomes. And CIK（cytokine induced killer cells）has been widely used in clinical treatment of cancers in China, and satisfactory results have been obtained. However, several problems remain to be solved: what kind of cells are suitable for adoptive transfer? How to efficiently induce and screen T cell clones? Whether and how to rapidly amplify T cells through TCR transgenic strategy? How to genetically engineer T cells to improve the efficacy of ACT, and how to properly evaluate the efficacy of ACT, etc.
Adoptive cell therapy (ACT) of cancer has long been proposed, but was gradually forgotten due to its disappointing efficacy. In recent years, great improvement has been made in cancer ACT．ACT with adoptive T cells for immunotherapy of melanoma has achieved impressive outcomes. And CIK（cytokine induced killer cells）has been widely used in clinical treatment of cancers in China, and satisfactory results have been obtained. However, several problems remain to be solved: what kind of cells are suitable for adoptive transfer? How to efficiently induce and screen T cell clones? Whether and how to rapidly amplify T cells through TCR transgenic strategy? How to genetically engineer T cells to improve the efficacy of ACT, and how to properly evaluate the efficacy of ACT, etc.
2011, 18(1):7-10.
DOI: 10.3872/j.issn.1007-385X.2011.1.002
Abstract:
Both the mechanisms and responses of tumor immunotherapy are different from those of the traditional therapy. Therefore, whether the traditional clinical response criteria are suitable for assessing the efficacy of immunotherapy is worth exploring. First, whether the immune response should be included in the response criteria of immunotherapy? Secondly, how to determine the time point for evaluating the efficacy of immunotherapy? Thirdly, which indices should be included for the new immune-related response criteria of tumor immunotherpay? Immunotherapy is showing an increasingly important role in clinical therapy of tumor, and it is of great significance to establish a scientific response criterion system for the development and clinical application of tumor immunotherapy.
Both the mechanisms and responses of tumor immunotherapy are different from those of the traditional therapy. Therefore, whether the traditional clinical response criteria are suitable for assessing the efficacy of immunotherapy is worth exploring. First, whether the immune response should be included in the response criteria of immunotherapy? Secondly, how to determine the time point for evaluating the efficacy of immunotherapy? Thirdly, which indices should be included for the new immune-related response criteria of tumor immunotherpay? Immunotherapy is showing an increasingly important role in clinical therapy of tumor, and it is of great significance to establish a scientific response criterion system for the development and clinical application of tumor immunotherapy.
2011, 18(1):11-16.
DOI: 10.3872/j.issn.1007-385X.2011.1.003
Abstract:
Objective:To screen for EGFRvⅢ specific single-chain Fv (scFv) by phage display library and to examine its targeting activity. Methods: EGFRvⅢ specific scFv phage library was constructed, and the positive EGFRvⅢ-scFv clone was screened by ELISA. After cloned into pCANTAB-Thrombin-His vector, EGFRvⅢ-scFv plasmid was transformed into E.coli HB2151, and soluble EGFRvⅢ-scFv was induced by IPTG. The specific binding activity of EGFRvⅢ-scFv with EGFRvⅢ was studied by indirect immunofluorescence and in vivo imaging. Results: An EGFRvⅢ-scFv phage library was successfully constructed and 16 EGFRvⅢ-scFv positive clones were identified by ELISA. One clone named EGFRvⅢ-scFv-2A1 was re-cloned into pCANTAB-Thrombin-His vector and soluble EGFRvⅢ-scFv-2A1 was successfully obtained. EGFRvⅢ-scFv-2A1 could specifically bind with HuH7-EGFRvⅢ and HuH7 hepatoma cells, but not with HuH7-EGFR and HuH7 cells in vitro. In vivo, fluorescence-labeled EGFRvⅢ-scFv-2A1 could only bind with U87MG-EGFRvⅢ glioma cells implanted tumor tissues, but not with that of U87MG cells implanted ones. Conclusion: The prepared EGFRvⅢ-scFv-2A1 can specifically bind with EGFRvⅢ, and it might be used for diagnosis and targeted therapy of tumors.
Objective:To screen for EGFRvⅢ specific single-chain Fv (scFv) by phage display library and to examine its targeting activity. Methods: EGFRvⅢ specific scFv phage library was constructed, and the positive EGFRvⅢ-scFv clone was screened by ELISA. After cloned into pCANTAB-Thrombin-His vector, EGFRvⅢ-scFv plasmid was transformed into E.coli HB2151, and soluble EGFRvⅢ-scFv was induced by IPTG. The specific binding activity of EGFRvⅢ-scFv with EGFRvⅢ was studied by indirect immunofluorescence and in vivo imaging. Results: An EGFRvⅢ-scFv phage library was successfully constructed and 16 EGFRvⅢ-scFv positive clones were identified by ELISA. One clone named EGFRvⅢ-scFv-2A1 was re-cloned into pCANTAB-Thrombin-His vector and soluble EGFRvⅢ-scFv-2A1 was successfully obtained. EGFRvⅢ-scFv-2A1 could specifically bind with HuH7-EGFRvⅢ and HuH7 hepatoma cells, but not with HuH7-EGFR and HuH7 cells in vitro. In vivo, fluorescence-labeled EGFRvⅢ-scFv-2A1 could only bind with U87MG-EGFRvⅢ glioma cells implanted tumor tissues, but not with that of U87MG cells implanted ones. Conclusion: The prepared EGFRvⅢ-scFv-2A1 can specifically bind with EGFRvⅢ, and it might be used for diagnosis and targeted therapy of tumors.
2011, 18(1):17-22.
DOI: 10.3872/j.issn.1007-385X.2011.1.004
Abstract:
Objective: To investigate the cytotoxicity and bystander effects of lentiviral vector-mediated yeast cytosine deaminase (YCD) suicide gene against the human leukemia Jurkat cells. Methods:YCD-GFP lentiviral vector was prepared and transfected into Jurkat cells (Jurkat/YCD-GFP cells). The expression of YCD-GFP gene was examined by flow cytometry. The cytotoxicity and bystander effects of pro-drug 5-fluorocytosine (5-FC) on Jurkat/YCD-GFP cells were measured in vitro. Nude mice bearing-Jurkat/YCD-GFP cell tumors were established and the cytotoxicity and bystander effects of 5-FC on tumor cells were measured in vivo. Results: YCD-GFP lentiviral vector and Jurkat/YCD-GFP cells (Jurkat cells infected by YCD-GFP lentiviral vector) were successfully prepared; the infection rate of YCD-GFP lentiviral vector on Jurkat cells was 96.93%. 5-FC (232 μmol/L for 72h) killed (95.61±2.07)% of Jurkat/YCD-GFP cells, and 5-FC level in the culture supernatant decreased by about 80%. Bystander effect of Jurkat/YCD-GFP cells could be observed both in mixed culture (effect∶target=1∶10, \[68.69± 4.97\]% vs \[97.87±1.11\]%, P<0.05) and in Transwell culture (effect∶target=1∶25, \[1.46±0.27\]×105 vs \[3.00±0.16\]×105，P<0.01) . In vivo experiment results demonstrated that YCD/5-FC suicide gene system had strong cytotoxicity and bystander effects in Jurkat/YCD-GFP tumor bearing mice (P<0.05). Conclusion: The lentiviral vector-mediated YCD/5-FC suicide gene system has marked cytotoxicity and bystander effects against human leukemia Jurkat cells.
Objective: To investigate the cytotoxicity and bystander effects of lentiviral vector-mediated yeast cytosine deaminase (YCD) suicide gene against the human leukemia Jurkat cells. Methods:YCD-GFP lentiviral vector was prepared and transfected into Jurkat cells (Jurkat/YCD-GFP cells). The expression of YCD-GFP gene was examined by flow cytometry. The cytotoxicity and bystander effects of pro-drug 5-fluorocytosine (5-FC) on Jurkat/YCD-GFP cells were measured in vitro. Nude mice bearing-Jurkat/YCD-GFP cell tumors were established and the cytotoxicity and bystander effects of 5-FC on tumor cells were measured in vivo. Results: YCD-GFP lentiviral vector and Jurkat/YCD-GFP cells (Jurkat cells infected by YCD-GFP lentiviral vector) were successfully prepared; the infection rate of YCD-GFP lentiviral vector on Jurkat cells was 96.93%. 5-FC (232 μmol/L for 72h) killed (95.61±2.07)% of Jurkat/YCD-GFP cells, and 5-FC level in the culture supernatant decreased by about 80%. Bystander effect of Jurkat/YCD-GFP cells could be observed both in mixed culture (effect∶target=1∶10, \[68.69± 4.97\]% vs \[97.87±1.11\]%, P<0.05) and in Transwell culture (effect∶target=1∶25, \[1.46±0.27\]×105 vs \[3.00±0.16\]×105，P<0.01) . In vivo experiment results demonstrated that YCD/5-FC suicide gene system had strong cytotoxicity and bystander effects in Jurkat/YCD-GFP tumor bearing mice (P<0.05). Conclusion: The lentiviral vector-mediated YCD/5-FC suicide gene system has marked cytotoxicity and bystander effects against human leukemia Jurkat cells.
2011, 18(1):23-27.
DOI: 10.3872/j.issn.1007-385X.2011.1.005
Abstract:
Objective:To observe the interaction of EGFR with IGFR-1β and to investigate the effect of EGFR inhibitor IGFR-1β on the anti-proliferative activity of gefitinib against colon carcinoma cells. Methods: Interactions between EGFR and IGFR-1β and their association with their downstream signal proteins AKT and MAPK were examined by immunoprecipitation and Western blotting analysis. MTT assay was used to measure cell proliferation of colon carcinoma cells after treatment with IGFR-1 tyrosine kinase inhibitor AG1024, gefitinib or both. Results: Irrespective of gefitinib treatment, EGFR immunoprecipitates from LoVo cells (the gefitinib-sensitive cell line) failed to display IGFR-1β band, whereas HCT116 cells (the gefitinib-resistant cell line) displayed the band and the band became more obvious after treatment with gefitinib. And IGFR-1β band was seen in gefitinib-treated HT29 cells (the gefitinib-moderate-sensitive cell line) but not in non-treated ones. Decreases in Akt and MAPK binding were found in LoVo cells treated with gefitinib, in HT29 cells treated with gefitinib plus AG1024, and in HCT116 cells treated with AG1024 (P<0.05). Gefitinib alone significantly decreased the proliferation rate of LoVo cells compared with control group (P<0.05), and AG1024 alone did not decrease the proliferation rate of LoVo cells; furthermore, gefitinib combined with AG1024 failed to further decrease the proliferation rate of LoVo cells compared with gefitinib group; AG1024 or gefitinib treatment alone did not decrease the proliferation rate of HT29 cells, but combined treatment led to a significant decrease (P<0.05); and AG1024 but not gefitinib alone significantly reduced the proliferation rate of HCT116 cells (P<0.05), whereas combined treatment did not result in a synergistic effect compared with AG1024 group. Conclusion: Resistance of colon carcinoma cells to gefitinib might partly arise from activation of IGFR-1β signaling pathway through the formation of EGFR/IGFR-1β heterodimerization, and administration of AG1024 blocking IGFR-1β activation might increase the sensitivity of colon carcinoma cells to gefitinib.
Objective:To observe the interaction of EGFR with IGFR-1β and to investigate the effect of EGFR inhibitor IGFR-1β on the anti-proliferative activity of gefitinib against colon carcinoma cells. Methods: Interactions between EGFR and IGFR-1β and their association with their downstream signal proteins AKT and MAPK were examined by immunoprecipitation and Western blotting analysis. MTT assay was used to measure cell proliferation of colon carcinoma cells after treatment with IGFR-1 tyrosine kinase inhibitor AG1024, gefitinib or both. Results: Irrespective of gefitinib treatment, EGFR immunoprecipitates from LoVo cells (the gefitinib-sensitive cell line) failed to display IGFR-1β band, whereas HCT116 cells (the gefitinib-resistant cell line) displayed the band and the band became more obvious after treatment with gefitinib. And IGFR-1β band was seen in gefitinib-treated HT29 cells (the gefitinib-moderate-sensitive cell line) but not in non-treated ones. Decreases in Akt and MAPK binding were found in LoVo cells treated with gefitinib, in HT29 cells treated with gefitinib plus AG1024, and in HCT116 cells treated with AG1024 (P<0.05). Gefitinib alone significantly decreased the proliferation rate of LoVo cells compared with control group (P<0.05), and AG1024 alone did not decrease the proliferation rate of LoVo cells; furthermore, gefitinib combined with AG1024 failed to further decrease the proliferation rate of LoVo cells compared with gefitinib group; AG1024 or gefitinib treatment alone did not decrease the proliferation rate of HT29 cells, but combined treatment led to a significant decrease (P<0.05); and AG1024 but not gefitinib alone significantly reduced the proliferation rate of HCT116 cells (P<0.05), whereas combined treatment did not result in a synergistic effect compared with AG1024 group. Conclusion: Resistance of colon carcinoma cells to gefitinib might partly arise from activation of IGFR-1β signaling pathway through the formation of EGFR/IGFR-1β heterodimerization, and administration of AG1024 blocking IGFR-1β activation might increase the sensitivity of colon carcinoma cells to gefitinib.
2011, 18(1):28-32.
DOI: 10.3872/j.issn.1007-385X.2011.1.006
Abstract:
Objective:To study the inhibitory effects of Xiaoai Jiedu Recipe(XJR) against transplanted hepatocarcinoma H22 tumors in mice and the related mechanism. Methods: H22-transplanted tumor mouse models were established and were divided into control group, XJR treatment groups (10, 30, 90 g/kg), and cisplatin treatment group. The growth of tumors in different groups was measured, and pathology changes of transplanted tumor tissues in different groups were examined by H-E staining. Flow cytometry was applied to examine cell cycle and sub-diploid apoptosis rate of transplanted tumor cells, and VEGF levels in the peripheral blood of mice were measured by ELISA. Results: Compared with control group, different XJR groups and the cisplatin group significantly inhibited growth of H22-transplanted tumors (245%, 42.8%, 21.1, 58.6% vs 0, P<0.05, P<0.01), cells in XJR groups showing massive necrosis and obvious chromosome condensation. Medium-dosage XJR and cisplatin blocked transplanted tumor cells in G0/G1 phase (P<005), and the cells in S phase were significantly reduced (P<0.05). The sub-diploid apoptosis rate of transplanted tumor cells in medium-doseage XJR group was similar to that in cisplatin group (60.52±6.40 vs 71.32±16.02, P>0.05). Peripheral VEGF levels were significantly lower in the medium-, large-dosage XJR groups and cisplatin group than in the control group (104.3±6.1, 105.8±7.2, 88.6±4.3 vs 120.7±12.6, P<0.05, P<0.01). Conclusion: Xiaoai Jiedu Recipe can inhibit the growth and promote apoptosis of H22-transplanted tumor cells, and inhibition of VEGF may be one of the mechanisms for the anticancer effect.
Objective:To study the inhibitory effects of Xiaoai Jiedu Recipe(XJR) against transplanted hepatocarcinoma H22 tumors in mice and the related mechanism. Methods: H22-transplanted tumor mouse models were established and were divided into control group, XJR treatment groups (10, 30, 90 g/kg), and cisplatin treatment group. The growth of tumors in different groups was measured, and pathology changes of transplanted tumor tissues in different groups were examined by H-E staining. Flow cytometry was applied to examine cell cycle and sub-diploid apoptosis rate of transplanted tumor cells, and VEGF levels in the peripheral blood of mice were measured by ELISA. Results: Compared with control group, different XJR groups and the cisplatin group significantly inhibited growth of H22-transplanted tumors (245%, 42.8%, 21.1, 58.6% vs 0, P<0.05, P<0.01), cells in XJR groups showing massive necrosis and obvious chromosome condensation. Medium-dosage XJR and cisplatin blocked transplanted tumor cells in G0/G1 phase (P<005), and the cells in S phase were significantly reduced (P<0.05). The sub-diploid apoptosis rate of transplanted tumor cells in medium-doseage XJR group was similar to that in cisplatin group (60.52±6.40 vs 71.32±16.02, P>0.05). Peripheral VEGF levels were significantly lower in the medium-, large-dosage XJR groups and cisplatin group than in the control group (104.3±6.1, 105.8±7.2, 88.6±4.3 vs 120.7±12.6, P<0.05, P<0.01). Conclusion: Xiaoai Jiedu Recipe can inhibit the growth and promote apoptosis of H22-transplanted tumor cells, and inhibition of VEGF may be one of the mechanisms for the anticancer effect.
2011, 18(1):33-37.
DOI: 10.3872/j.issn.1007-385X.2011.1.007
Abstract:
Objective:To study the influence of norcantharidin (NCTD) on the proliferation, apoptosis and survivin expression of human hepatoma HepG2 cells. Methods: MTT assay was used to investigate the effect of NCTD on proliferation of HepG2 cells; Hochest 33258 staining, Annenxin V/PI staining, and agarose gel electrophoresis assay were used to examine the effect of NCTD on apoptosis of HepG2 cells. Western blotting analysis was used to examine the effect of NCTD on survivin expression in HepG2 cells. Results: Proliferation of HepG2 cells was markedly inhibited by NCTD in a dose-dependent manner (5, 10, 20, 40 μg/ml), with the inhibitory rate being (81.27±3.25)% after 40 μg/ml NCTD treatment for 48 h. After HepG2 cells were treated with NCTD for 24 h, fuorescence microscopy revealed that the cell nuclei were condensed and fragmented, and agarose gel electrophoresis of DNA showed a typical DNA ladder map of apoptosis. Flow cytometry results showed that the apoptosis rates of HepG2 cells were (7.33±0.25)%, (18.23±119)%, (32.5±2.30)%, and (48.23±1.17)% after 5, 10, 20, and 40 μg/ml NCTD treatments, respectively. Western blotting analysis results demonstrated that the survivin expression in HepG2 cells was significantly down-regulated with the increase of NCTD dosage. Conclusion: NCTD can inhibit proliferation and induce apoptosis of HepG2 cells, which is related to the down-regulation of survivin protein.
Objective:To study the influence of norcantharidin (NCTD) on the proliferation, apoptosis and survivin expression of human hepatoma HepG2 cells. Methods: MTT assay was used to investigate the effect of NCTD on proliferation of HepG2 cells; Hochest 33258 staining, Annenxin V/PI staining, and agarose gel electrophoresis assay were used to examine the effect of NCTD on apoptosis of HepG2 cells. Western blotting analysis was used to examine the effect of NCTD on survivin expression in HepG2 cells. Results: Proliferation of HepG2 cells was markedly inhibited by NCTD in a dose-dependent manner (5, 10, 20, 40 μg/ml), with the inhibitory rate being (81.27±3.25)% after 40 μg/ml NCTD treatment for 48 h. After HepG2 cells were treated with NCTD for 24 h, fuorescence microscopy revealed that the cell nuclei were condensed and fragmented, and agarose gel electrophoresis of DNA showed a typical DNA ladder map of apoptosis. Flow cytometry results showed that the apoptosis rates of HepG2 cells were (7.33±0.25)%, (18.23±119)%, (32.5±2.30)%, and (48.23±1.17)% after 5, 10, 20, and 40 μg/ml NCTD treatments, respectively. Western blotting analysis results demonstrated that the survivin expression in HepG2 cells was significantly down-regulated with the increase of NCTD dosage. Conclusion: NCTD can inhibit proliferation and induce apoptosis of HepG2 cells, which is related to the down-regulation of survivin protein.
2011, 18(1):38-41.
DOI: 10.3872/j.issn.1007-385X.2011.1.008
Abstract:
Objective:To study the effect of Snail on resistance of breast cancer MCF-7 cell transplanted tumors to doxorubicin and its possible mechanism. Methods: Snail eukaryotic expression vector pcDNA3.1-Snail was constructed and transfected into MCF-7 cells, and MCF-7 cells with stable Snail expression (MCF-7/Snail cells) were screened. MCF-7 cells transfected with blank pcDNA3.1 (MCF-7/pcDNA cells) were used as control. MCF-7/Snail- and MCF-7/pcDNA-cell transplanted tumor models were established. After doxorubicin injection, the growth of transplanted tumors was observed, and the inhibitory rate of doxorubicin was calculated. The expressions of Snail, MDR-1 and MMP-9 in transplanted tumor tissues were examined by immunohistochemistry. Results: pcDNA3.1-Snail expression vector was successfully constructed, and MCF-7/Snail and MCF-7/pcDNA cells were obtained. After doxorubicin therapy, the transplanted tumor weight in MCF-7/Snail group was significantly higher than that in the MCF-7/pcDNA group (\[1.413±0674\]g vs \[1.257±0576\]g, P<0.05), and the inhibitory rate of doxorubicin was significantly lower (18.42% vs 30.18%, P<0.05). The expressions of Snail, MDR-1 and MMP-9 in transplanted tumor tissues were significantly higher than those in MCF-7/pcDNA group (408.08±20.39 vs 67.67±16.56, 363.50±26.56 vs 55.08±12.23, 396.25±16.03 vs 56.92±7.35, all P<0.05)，and the expression of Snail was positively correlated with that of MDR-1 and MMP-9 (r1=0.89, P<0.01; r2=0.81, P<0.01). Conclusion:Snail can increase resistance of breast cancer MCF-7 cell transplanted tumors to doxorubicin, which might be related with the increased expressions of MDR-1 and MMP-9 in breast cancer MCF-7 transplanted tumors.
Objective:To study the effect of Snail on resistance of breast cancer MCF-7 cell transplanted tumors to doxorubicin and its possible mechanism. Methods: Snail eukaryotic expression vector pcDNA3.1-Snail was constructed and transfected into MCF-7 cells, and MCF-7 cells with stable Snail expression (MCF-7/Snail cells) were screened. MCF-7 cells transfected with blank pcDNA3.1 (MCF-7/pcDNA cells) were used as control. MCF-7/Snail- and MCF-7/pcDNA-cell transplanted tumor models were established. After doxorubicin injection, the growth of transplanted tumors was observed, and the inhibitory rate of doxorubicin was calculated. The expressions of Snail, MDR-1 and MMP-9 in transplanted tumor tissues were examined by immunohistochemistry. Results: pcDNA3.1-Snail expression vector was successfully constructed, and MCF-7/Snail and MCF-7/pcDNA cells were obtained. After doxorubicin therapy, the transplanted tumor weight in MCF-7/Snail group was significantly higher than that in the MCF-7/pcDNA group (\[1.413±0674\]g vs \[1.257±0576\]g, P<0.05), and the inhibitory rate of doxorubicin was significantly lower (18.42% vs 30.18%, P<0.05). The expressions of Snail, MDR-1 and MMP-9 in transplanted tumor tissues were significantly higher than those in MCF-7/pcDNA group (408.08±20.39 vs 67.67±16.56, 363.50±26.56 vs 55.08±12.23, 396.25±16.03 vs 56.92±7.35, all P<0.05)，and the expression of Snail was positively correlated with that of MDR-1 and MMP-9 (r1=0.89, P<0.01; r2=0.81, P<0.01). Conclusion:Snail can increase resistance of breast cancer MCF-7 cell transplanted tumors to doxorubicin, which might be related with the increased expressions of MDR-1 and MMP-9 in breast cancer MCF-7 transplanted tumors.
2011, 18(1):42-45.
DOI: 10.3872/j.issn.1007-385X.2011.1.009
Abstract:
Objective:To prepare the TAT-ASPP2 fusion protein and investigate its inhibitory effect against the proliferation of glioma U-87MG and U251 cells. Methods: TAT-ASPP2 specific primer was designed and recombinant prokaryotic expression vector pET-TAT-ASPP2 was constructed using the In-Fusion cloning technique. After identified by double endonuclease digestion and DNA sequencing, pET-TAT-ASPP2 vector was transformed into E. coli BL21 and TAT-ASPP2 fusion protein was induced by IPTG. TAT-ASPP2 fusion protein was further identified by SDS-PACE and Western blotting analysis. The effects of TAT-ASPP2 fusion protein on proliferation of U-87MG and U251 cells were detected by MTT assay. Results: The prokaryotic expression plasmid pET-TAT-ASPP2 was successfully constructed, and TAT-ASPP2 fusion protein was induced by IPTG in transformed E.coli BL21; the molecular weight of the fusion protein was about 128 000 and it could be specifically recognized by ASPP2 antibody. TAT-ASPP2 fusion protein significantly inhibited the proliferation of U-87MG and U-251 cells, with the inhibitory rates being about (65.0±3.0)% and (64.7±2.5)%, respectively; while ASPP2 protein did not inhibit the proliferation of U-87MG and U-251 cells. Conclusion: TAT-ASPP2 fusion protein has been successfully expressed and purified, and the fusion protein can significantly inhibit the proliferation of glioma cells.
Objective:To prepare the TAT-ASPP2 fusion protein and investigate its inhibitory effect against the proliferation of glioma U-87MG and U251 cells. Methods: TAT-ASPP2 specific primer was designed and recombinant prokaryotic expression vector pET-TAT-ASPP2 was constructed using the In-Fusion cloning technique. After identified by double endonuclease digestion and DNA sequencing, pET-TAT-ASPP2 vector was transformed into E. coli BL21 and TAT-ASPP2 fusion protein was induced by IPTG. TAT-ASPP2 fusion protein was further identified by SDS-PACE and Western blotting analysis. The effects of TAT-ASPP2 fusion protein on proliferation of U-87MG and U251 cells were detected by MTT assay. Results: The prokaryotic expression plasmid pET-TAT-ASPP2 was successfully constructed, and TAT-ASPP2 fusion protein was induced by IPTG in transformed E.coli BL21; the molecular weight of the fusion protein was about 128 000 and it could be specifically recognized by ASPP2 antibody. TAT-ASPP2 fusion protein significantly inhibited the proliferation of U-87MG and U-251 cells, with the inhibitory rates being about (65.0±3.0)% and (64.7±2.5)%, respectively; while ASPP2 protein did not inhibit the proliferation of U-87MG and U-251 cells. Conclusion: TAT-ASPP2 fusion protein has been successfully expressed and purified, and the fusion protein can significantly inhibit the proliferation of glioma cells.
2011, 18(1):46-50.
DOI: 10.3872/j.issn.1007-385X.2011.1.010
Abstract:
Objective:To investigate the effect of RNA-mediated interference of hTERT (human telomerase reverse transcriptase) expression on the apoptosis of colorectal cancer cell line SW480. Methods：Small hairpin RNA (shRNA) targeting hTERT was synthesized and recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA (named hTERT-shRNA plasmid) containing hTERT-shRNA was constructed. SW480 cells were transfected with hTERT-shRNA plasmid by liposome method, and the expression of hTERT mRNA in SW480 cells was detected by RT-PCR analysis at different time points. The telomerase activity of SW480 cells was examined by TRAP-PCR-ELISA analysis. The ultrastructure of SW480 cells was examined by TEM (transparent electron microscope) 48 h after hTERT-shRNA transfection. Results: The inhibitory rate of hTERT mRNA expression in SW480 cells of hTERT-shRNA group was significantly higher than those of blank group, liposome group, and NC-shRNA group (75.0% vs 39.2%, 33.3%, 28.0%, P<0.05). Telomerase activity in SW480 cells of hTERT-shRNA group was significantly decreased compared with those of blank group, liposome group, and NC-shRNA group (2.242±0.285 vs 2.756±0.089, 2.693±0.225, 2.691±0.120, P<0.05). SW480 cells in hTERT-shRNA group showed smaller cell size, nuclear condensation, uneven aggregation of chromatin along the nuclear membrane, and increased vacuolization. Conclusion：RNA interference can effectively silence hTERT expression, reduce telomerase activity, and induce apoptosis of SW480 cells.
Objective:To investigate the effect of RNA-mediated interference of hTERT (human telomerase reverse transcriptase) expression on the apoptosis of colorectal cancer cell line SW480. Methods：Small hairpin RNA (shRNA) targeting hTERT was synthesized and recombinant plasmid pGPU6/GFP/Neo-hTERT-shRNA (named hTERT-shRNA plasmid) containing hTERT-shRNA was constructed. SW480 cells were transfected with hTERT-shRNA plasmid by liposome method, and the expression of hTERT mRNA in SW480 cells was detected by RT-PCR analysis at different time points. The telomerase activity of SW480 cells was examined by TRAP-PCR-ELISA analysis. The ultrastructure of SW480 cells was examined by TEM (transparent electron microscope) 48 h after hTERT-shRNA transfection. Results: The inhibitory rate of hTERT mRNA expression in SW480 cells of hTERT-shRNA group was significantly higher than those of blank group, liposome group, and NC-shRNA group (75.0% vs 39.2%, 33.3%, 28.0%, P<0.05). Telomerase activity in SW480 cells of hTERT-shRNA group was significantly decreased compared with those of blank group, liposome group, and NC-shRNA group (2.242±0.285 vs 2.756±0.089, 2.693±0.225, 2.691±0.120, P<0.05). SW480 cells in hTERT-shRNA group showed smaller cell size, nuclear condensation, uneven aggregation of chromatin along the nuclear membrane, and increased vacuolization. Conclusion：RNA interference can effectively silence hTERT expression, reduce telomerase activity, and induce apoptosis of SW480 cells.
2011, 18(1):51-54.
DOI: 10.3872/j.issn.1007-385X.2011.1.011
Abstract:
Objective:To investigate the effects of nerifolin, an isolate from the seeds of Cerbera manghas L., on the proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2 and the related mechanism. Methods: Effect of nerifolin on the proliferation of HepG2 cells was examined by MTT assay, and its effects on cell cycle and apoptosis of HepG2 cells were assessed by flow cytometry. Effect of nerifolin on caspase-3 activation in HepG2 cells was examined by caspase detecting kit. Results: Nerifolin inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner, with the IC50 values being (2.34±0.08), (0.13±0.01) and (0.06±0.01) μg/ml after nerifolin treatment for 24, 48, and 72 h, respectively. The proportion of HepG2 cells in S phase increased with the prolongation of nerifolin treatment, and the proportion in G0/G1 phase gradually decreased (P<0.05), indicating that nerifolin blocked HepG2 cells in S phase. Early apoptosis rate of HepG2 cells increased to 22.65%, and caspase-3 activity was significantly increased after nerifolin treatment (P<0.01). Conclusion: Nerifolin can inhibit the proliferation of HepG2 cells by inducing S phase arrest and can trigger apoptosis via caspase-3 dependent pathway.
Objective:To investigate the effects of nerifolin, an isolate from the seeds of Cerbera manghas L., on the proliferation and apoptosis of human hepatocellular carcinoma cell line HepG2 and the related mechanism. Methods: Effect of nerifolin on the proliferation of HepG2 cells was examined by MTT assay, and its effects on cell cycle and apoptosis of HepG2 cells were assessed by flow cytometry. Effect of nerifolin on caspase-3 activation in HepG2 cells was examined by caspase detecting kit. Results: Nerifolin inhibited the proliferation of HepG2 cells in a dose- and time-dependent manner, with the IC50 values being (2.34±0.08), (0.13±0.01) and (0.06±0.01) μg/ml after nerifolin treatment for 24, 48, and 72 h, respectively. The proportion of HepG2 cells in S phase increased with the prolongation of nerifolin treatment, and the proportion in G0/G1 phase gradually decreased (P<0.05), indicating that nerifolin blocked HepG2 cells in S phase. Early apoptosis rate of HepG2 cells increased to 22.65%, and caspase-3 activity was significantly increased after nerifolin treatment (P<0.01). Conclusion: Nerifolin can inhibit the proliferation of HepG2 cells by inducing S phase arrest and can trigger apoptosis via caspase-3 dependent pathway.
2011, 18(1):55-58.
DOI: 10.3872/j.issn.1007-385X.2011.1.012
Abstract:
Objective:To study the efficacy of mixed-HSCT (autologous peripheral blood stem cell mixed with HLA haploidentical allogeneic bone marrow transplantation) combined with donor lymphocyte infusion plus interleukin-2 (DLI+IL-2) in treatment of acute myelogenous leukemia (AML) patients. Methods: Twenty-three AML patients (15 males and 8 females, median age 22 years) were enrolled in this study as mixed-HSCT combined DLI+IL-2 group, and 14 AML patients (10 males and 4 females, median age 21 years) were enrolled as control group. All patients in the two groups received TBI+VEMAC therapy after complete remission, patients in the control group were treated with mixed-HSCT alone, and those in the experimental group were treated with Mixed-HSCT and further treatment with DLI+IL-2 for 1-8 times after hematopoietic reconstruction. All the patients were followed up for at least 3 years. Results: All patients in the two groups achieved hematopoietic reconstruction and had no graft-versus-host disease (GVHD). During 3 years, 6 cases in the experimental group formed mixed chimerism (46XX/46XY) and 15 survived, with the disease-free survival rate (DFS) being 65.2%. Three cases in the control group formed mixed chimerism (46XX/46XY) and 7 cases survived, with the DFS being 50.0%. All patients in the two groups had similar adverse reactions (oral cavity ulcer, hemorrhagic cystitis, fever, etc.) after therapy. Conclusion: Mixed-HSCT combined with DLI+IL-2 has a positive effect on DFS of AML patients, with no noticeable adverse reactions.
Objective:To study the efficacy of mixed-HSCT (autologous peripheral blood stem cell mixed with HLA haploidentical allogeneic bone marrow transplantation) combined with donor lymphocyte infusion plus interleukin-2 (DLI+IL-2) in treatment of acute myelogenous leukemia (AML) patients. Methods: Twenty-three AML patients (15 males and 8 females, median age 22 years) were enrolled in this study as mixed-HSCT combined DLI+IL-2 group, and 14 AML patients (10 males and 4 females, median age 21 years) were enrolled as control group. All patients in the two groups received TBI+VEMAC therapy after complete remission, patients in the control group were treated with mixed-HSCT alone, and those in the experimental group were treated with Mixed-HSCT and further treatment with DLI+IL-2 for 1-8 times after hematopoietic reconstruction. All the patients were followed up for at least 3 years. Results: All patients in the two groups achieved hematopoietic reconstruction and had no graft-versus-host disease (GVHD). During 3 years, 6 cases in the experimental group formed mixed chimerism (46XX/46XY) and 15 survived, with the disease-free survival rate (DFS) being 65.2%. Three cases in the control group formed mixed chimerism (46XX/46XY) and 7 cases survived, with the DFS being 50.0%. All patients in the two groups had similar adverse reactions (oral cavity ulcer, hemorrhagic cystitis, fever, etc.) after therapy. Conclusion: Mixed-HSCT combined with DLI+IL-2 has a positive effect on DFS of AML patients, with no noticeable adverse reactions.
2011, 18(1):59-62.
DOI: 10.3872/j.issn.1007-385X.2011.1.013
Abstract:
Objective:To study the ratio of Th17 cells in peripheral lymphocytes and the expression of RORγt and IL-17 in the peripheral blood mononuclear cells (PBMCs) of patients with esophageal squamous cell carcinoma (ESCC), and to investigate their clinical significance. Methods: Forty PBMC samples from ESCC patients (24 men and 16 women, mean age 61±8.76 years), who were diagnosed in Qilu Hospital of Shandong University from August 2009 to June 2010, and 40 healthy volunteers were included in the present study. The ratio of Th17 cells in peripheral lymphocytes was detected by flow cytometry, and the expression levels of IL-17 and RORγt mRNA in PBMC were examined by PT-PCR. Results: The ratio of Th17 cells in the peripheral lymphocytes of ESCC patients was significantly higher than that in healthy volunteers (\[2.40±0.55\] % vs \[0.84±0.41\]%, P<0. 01). Both RORγt and IL-17 mRNA expression levels in ESCC patients were significantly higher than those in the healthy volunteers (\[0.669±0.184\] vs \[0.451士0.151\], \[0.625±0.179\] vs \[0.438±0.150\], P<0.01), and the two showed a positive correlation (r2 =0.551, P<0.01). In addition, the Th17 ratio, and RORγt and IL-17 mRNA expression levels were significantly higher in metastasis ESCC patients compared with those in the non-metastasis ESCC patients (P<0.05). Conclusion: Th17 ratio and RORγt and IL-17 mRNA expression levels are significantly increased in ESCC patients, and Th17 cells may participate in the development of ESCC through RORγt and IL-17.
Objective:To study the ratio of Th17 cells in peripheral lymphocytes and the expression of RORγt and IL-17 in the peripheral blood mononuclear cells (PBMCs) of patients with esophageal squamous cell carcinoma (ESCC), and to investigate their clinical significance. Methods: Forty PBMC samples from ESCC patients (24 men and 16 women, mean age 61±8.76 years), who were diagnosed in Qilu Hospital of Shandong University from August 2009 to June 2010, and 40 healthy volunteers were included in the present study. The ratio of Th17 cells in peripheral lymphocytes was detected by flow cytometry, and the expression levels of IL-17 and RORγt mRNA in PBMC were examined by PT-PCR. Results: The ratio of Th17 cells in the peripheral lymphocytes of ESCC patients was significantly higher than that in healthy volunteers (\[2.40±0.55\] % vs \[0.84±0.41\]%, P<0. 01). Both RORγt and IL-17 mRNA expression levels in ESCC patients were significantly higher than those in the healthy volunteers (\[0.669±0.184\] vs \[0.451士0.151\], \[0.625±0.179\] vs \[0.438±0.150\], P<0.01), and the two showed a positive correlation (r2 =0.551, P<0.01). In addition, the Th17 ratio, and RORγt and IL-17 mRNA expression levels were significantly higher in metastasis ESCC patients compared with those in the non-metastasis ESCC patients (P<0.05). Conclusion: Th17 ratio and RORγt and IL-17 mRNA expression levels are significantly increased in ESCC patients, and Th17 cells may participate in the development of ESCC through RORγt and IL-17.
2011, 18(1):63-69.
DOI: 10.3872/j.issn.1007-385X.2011.1.014
Abstract:
Objective:To systematically assess the efficacy and safety of rituximab combined with routine chemotherapy in the treatment of B-cell lymphoma by Meta-analysis. Methods: We searched Embase, Pubmed, the Cochrane Library, VIP, CNKI, and CBM literature databases for randomized controlled trials (RCTs) of rituximab combined with routine chemotherapy in the treatment of B-cell lymphoma. Two reviewers independently assessed the quality of the included studies and extracted the data. The data were analyzed by Review Manager software (version 5.0). Results: Ten RCTs were finally included in the present analysis, and the results showed that rituximab combined with routine chemotherapy improved the overall survival rate (HR=0.64, 95% CI \[0.53, 0.77\]) and overall response rate (RR=1.21, 95% CI \[1.11, 1.32\]) of B-cell lymphoma patients compared with routine chemotherapy. There was no statistical difference between the two groups in 3/4 grade infection, 3/4 grade thrombocytopenia, with the relative risk being 1.15 (0.58, 230) and 1.04 (0.71, 1.52), respectively. There was significant difference in 3/4 grade granulocytopenia, 3/4 grade granulocytopenia, 3/4 grade leukocytopenia, and 3/4 grade fever, with the relative risk being 1.16 (1.02, 1.31), 1.31 (112, 153) and 349 (1.56, 778), respectively. Conclusion: Rituximab combined with routine chemotherapy can improve the overall survival rate and overall response rate of B-cell lymphoma patients, but results in a higher incidence of 3/4 grade granulocytopenia, 3/4 grade granulocytopenia, 3/4 grade leukocytopenia, and 3/4 grade fever.
Objective:To systematically assess the efficacy and safety of rituximab combined with routine chemotherapy in the treatment of B-cell lymphoma by Meta-analysis. Methods: We searched Embase, Pubmed, the Cochrane Library, VIP, CNKI, and CBM literature databases for randomized controlled trials (RCTs) of rituximab combined with routine chemotherapy in the treatment of B-cell lymphoma. Two reviewers independently assessed the quality of the included studies and extracted the data. The data were analyzed by Review Manager software (version 5.0). Results: Ten RCTs were finally included in the present analysis, and the results showed that rituximab combined with routine chemotherapy improved the overall survival rate (HR=0.64, 95% CI \[0.53, 0.77\]) and overall response rate (RR=1.21, 95% CI \[1.11, 1.32\]) of B-cell lymphoma patients compared with routine chemotherapy. There was no statistical difference between the two groups in 3/4 grade infection, 3/4 grade thrombocytopenia, with the relative risk being 1.15 (0.58, 230) and 1.04 (0.71, 1.52), respectively. There was significant difference in 3/4 grade granulocytopenia, 3/4 grade granulocytopenia, 3/4 grade leukocytopenia, and 3/4 grade fever, with the relative risk being 1.16 (1.02, 1.31), 1.31 (112, 153) and 349 (1.56, 778), respectively. Conclusion: Rituximab combined with routine chemotherapy can improve the overall survival rate and overall response rate of B-cell lymphoma patients, but results in a higher incidence of 3/4 grade granulocytopenia, 3/4 grade granulocytopenia, 3/4 grade leukocytopenia, and 3/4 grade fever.
2011, 18(1):70-74.
DOI: 10.3872/j.issn.1007-385X.2011.1.015
Abstract:
Objective:To systematically evaluate the efficacy and safety of intravesical therapy with Bacillus Calmette-Guérin (BCG) following transurethral resection (TUR) in patients with superficial bladder cancer. Methods：We searched PubMed, EMBASE, the Cochrane Library, Chinese Biomedical Literature Database, Chinese Scientific Journal Full-text Database, and Chinese Journal Full-text Database for randomized controlled trials (RCTs) for studies about intravesical therapy with BCG following TUR in patients with superficial bladder cancer. Two reviewers independently evaluated the quality of the included studies and extracted the data. The data were analyzed using Review Manager 5.0 provided by Cochrane collaboration. Results：Five RCTs (totaling 450 patients) were finally included in the present study. The results showed that there were significant differences in recurrence rates (\[RR=0.64, 95% CI \[0.52, 0.80\]), cystitis ( RR=5063, 95% CI \[14.57, 175.90\]), haematuria (RR=18.28, 95% CI \[5.23, 63.85\]), fever ( RR=2193, 95% CI \[632, 76.02\] ), malaise (RR=10.23, 95% CI \[2.06, 50.81\]), and nausea (RR=4.67, 95% CI \[0.83, 2631\]) between the intravesical therapy with BCG following TUR group and TUR alone group, indicating that intravesical therapy with BCG following TUR was better than TUR alone in preventing the recurrence of bladder cancer, but with more adverse effects. Conclusion：Compared with TUR alone, intravesical therapy with BCG following TUR appears to decrease the tumor recurrence in patients with superficial bladder cancer, but with more adverse effects.
Objective:To systematically evaluate the efficacy and safety of intravesical therapy with Bacillus Calmette-Guérin (BCG) following transurethral resection (TUR) in patients with superficial bladder cancer. Methods：We searched PubMed, EMBASE, the Cochrane Library, Chinese Biomedical Literature Database, Chinese Scientific Journal Full-text Database, and Chinese Journal Full-text Database for randomized controlled trials (RCTs) for studies about intravesical therapy with BCG following TUR in patients with superficial bladder cancer. Two reviewers independently evaluated the quality of the included studies and extracted the data. The data were analyzed using Review Manager 5.0 provided by Cochrane collaboration. Results：Five RCTs (totaling 450 patients) were finally included in the present study. The results showed that there were significant differences in recurrence rates (\[RR=0.64, 95% CI \[0.52, 0.80\]), cystitis ( RR=5063, 95% CI \[14.57, 175.90\]), haematuria (RR=18.28, 95% CI \[5.23, 63.85\]), fever ( RR=2193, 95% CI \[632, 76.02\] ), malaise (RR=10.23, 95% CI \[2.06, 50.81\]), and nausea (RR=4.67, 95% CI \[0.83, 2631\]) between the intravesical therapy with BCG following TUR group and TUR alone group, indicating that intravesical therapy with BCG following TUR was better than TUR alone in preventing the recurrence of bladder cancer, but with more adverse effects. Conclusion：Compared with TUR alone, intravesical therapy with BCG following TUR appears to decrease the tumor recurrence in patients with superficial bladder cancer, but with more adverse effects.
LI Xue-yuan
,
JIA Ai-hua
,
REN Yu-bo
,
LI Xin
,
MA Xiang-yu
,
ZHANG Lian-qun
,
ZHANG Xue-guang
,
LI Xin-gang
2011, 18(1):75-79.
DOI: 10.3872/j.issn.1007-385X.2011.1.016
Abstract:
Objective:To study the expression of mammalian target of rapamycin (mTOR) signaling pathway protein in glioma patients and to discuss the related clinical significances. Methods: Eighty-seven paraffin samples from glioma patients (27 of Ⅰ-Ⅱ grade, 24 of Ⅲ grade, 24 of Ⅳ grade), who were pathologically diagnosed in Department of Neurological Surgery, Qilu Hospital and People’s Hospital of Liaocheng from Jul. 2007 to May. 2010, were obtained. Immunohistochemistry was used to examine the expression levels of 3 key proteins of mTOR signaling pathway, pAKT, pmTOR and p-p70S6K, in the glioma tissues, and their relationship with clinicopathological characteristics of glioma patients was discussed. Results: The positive rates of pAKT, pmTOR and p-p70S6K in different grades of glioma tissues showed no significant difference (P>0.05), with pAKT being 70.4% (19/27) inⅠ-Ⅱ grade, 70.8% (17/24) in Ⅲ grade, 75% (27/36) in Ⅳ grade; pmTOR being 77.8% (21/27) inⅠ-Ⅱ grade, 75.0% (18/24) in Ⅲ grade, 722% (26/36) in Ⅳ grade; and p-p70S6K being 63% (17/27) inⅠ-Ⅱ grade, 70.8% (17/24) in Ⅲ grade, 806% (29/36) in Ⅳ grade tissues. The positive rates increased with the increase of pathological grades (P=0.0001, 0.0063, 0.0001, respectively), and they showed no relationship with gender, age, tumor volume, or KPS score of glioma patients. pAKT, pmTOR and p-p70S6K co-expressed in 42 cases; only one or two proteins were co-expressed in another 45 cases. Conclusion: mTOR signaling pathway proteins may play important roles in the development and progression of glioma, and they may serve as potential targets in treatment of glioma.
Objective:To study the expression of mammalian target of rapamycin (mTOR) signaling pathway protein in glioma patients and to discuss the related clinical significances. Methods: Eighty-seven paraffin samples from glioma patients (27 of Ⅰ-Ⅱ grade, 24 of Ⅲ grade, 24 of Ⅳ grade), who were pathologically diagnosed in Department of Neurological Surgery, Qilu Hospital and People’s Hospital of Liaocheng from Jul. 2007 to May. 2010, were obtained. Immunohistochemistry was used to examine the expression levels of 3 key proteins of mTOR signaling pathway, pAKT, pmTOR and p-p70S6K, in the glioma tissues, and their relationship with clinicopathological characteristics of glioma patients was discussed. Results: The positive rates of pAKT, pmTOR and p-p70S6K in different grades of glioma tissues showed no significant difference (P>0.05), with pAKT being 70.4% (19/27) inⅠ-Ⅱ grade, 70.8% (17/24) in Ⅲ grade, 75% (27/36) in Ⅳ grade; pmTOR being 77.8% (21/27) inⅠ-Ⅱ grade, 75.0% (18/24) in Ⅲ grade, 722% (26/36) in Ⅳ grade; and p-p70S6K being 63% (17/27) inⅠ-Ⅱ grade, 70.8% (17/24) in Ⅲ grade, 806% (29/36) in Ⅳ grade tissues. The positive rates increased with the increase of pathological grades (P=0.0001, 0.0063, 0.0001, respectively), and they showed no relationship with gender, age, tumor volume, or KPS score of glioma patients. pAKT, pmTOR and p-p70S6K co-expressed in 42 cases; only one or two proteins were co-expressed in another 45 cases. Conclusion: mTOR signaling pathway proteins may play important roles in the development and progression of glioma, and they may serve as potential targets in treatment of glioma.
2011, 18(1):80-83.
DOI: 10.3872/j.issn.1007-385X.2011.1.017
Abstract:
Objective:To investigate the expression of Snail protein in human glioma tissues and study the effect of Snail on human glioma U251 cells. Methods: Sixty-five specimens from glioma patients, who were diagnosed in Beijing Anzhen Hospital and Beijing Tiantan Hospital, were included in this study. Immunohistochemistry S-P was used to detect Snail protein expression in human glioma tissues. Snail specific small interference RNA (Snail-siRNA) was constructed and transfected into U251 cells by lipofectamine. The expressions of Snail mRNA and protein and E-cadherin protein in transfected-U251 cells were investigated by RT-PCR and Western blotting analysis, respectively; and the invasion ability of transfected-U251 cells was investigated by Transwell chamber assay. Results: The positive rate of Snail was 662% in human glioma, which was significantly higher than that in the normal brain tissues (0%, P<0.01), and Snail positive rate in Ⅰ-Ⅱ stage glioma was significantly lower than that in Ⅲ-Ⅳ stage glioma (44.8% vs 83.3%, P<001). Snail-siRNA transfection inhibited the expressions of Snail mRNA and protein. The expression of E-cadherin protein in Snail-siRNA transfected-U251 cells was significantly increased compared with those in Ctrl-siRNA and untransfected groups (064±0.21 vs 0.15±0.16, 0.21±0.19, P<0.01). Snail-siRNA transfection inhibited the invasion of U251 cells （87.0±2.4 vs 140.0±4.9, 136.0±5.3; P<0.05）.Conclusion: Snail protein is highly expressed in human glioma tissues, and siRNA interfering the expression of Snail protein can inhibit invasion of glioma U251 cells.
Objective:To investigate the expression of Snail protein in human glioma tissues and study the effect of Snail on human glioma U251 cells. Methods: Sixty-five specimens from glioma patients, who were diagnosed in Beijing Anzhen Hospital and Beijing Tiantan Hospital, were included in this study. Immunohistochemistry S-P was used to detect Snail protein expression in human glioma tissues. Snail specific small interference RNA (Snail-siRNA) was constructed and transfected into U251 cells by lipofectamine. The expressions of Snail mRNA and protein and E-cadherin protein in transfected-U251 cells were investigated by RT-PCR and Western blotting analysis, respectively; and the invasion ability of transfected-U251 cells was investigated by Transwell chamber assay. Results: The positive rate of Snail was 662% in human glioma, which was significantly higher than that in the normal brain tissues (0%, P<0.01), and Snail positive rate in Ⅰ-Ⅱ stage glioma was significantly lower than that in Ⅲ-Ⅳ stage glioma (44.8% vs 83.3%, P<001). Snail-siRNA transfection inhibited the expressions of Snail mRNA and protein. The expression of E-cadherin protein in Snail-siRNA transfected-U251 cells was significantly increased compared with those in Ctrl-siRNA and untransfected groups (064±0.21 vs 0.15±0.16, 0.21±0.19, P<0.01). Snail-siRNA transfection inhibited the invasion of U251 cells （87.0±2.4 vs 140.0±4.9, 136.0±5.3; P<0.05）.Conclusion: Snail protein is highly expressed in human glioma tissues, and siRNA interfering the expression of Snail protein can inhibit invasion of glioma U251 cells.
2011, 18(1):89-91.
DOI: 10.3872/j.issn.1007-385X.2011.1.019
Abstract:
目的：探讨乳腺癌组织中GATA3的表达及与ER表达的关系，以及GATA3在乳腺癌发生中的意义。方法：采用免疫组化方法检测了上海市黄浦区中心医院2004年至2005年间外科手术切除的100例乳腺癌组织和61例癌旁乳腺组织中GATA3的表达水平，分析其与ER表达和乳腺癌患者临床病理特征间的关系。结果：GATA3在癌旁乳腺组织中的阳性表达率（82.0%）明显高于乳腺癌组织（60.0%）（χ2＝8.45，P<0.01），但在不同病理类型乳腺癌中表达无差异（χ2＝0.74，P>005）GATA3的表达和患者年龄、肿瘤大小无关（P>0.05），与乳腺癌组织分级、淋巴结转移相关（P<0.05）。乳腺癌组织中GATA3的阳性表达与ER的阳性表达存在着显著的正相关（r=0.49，P<0.01）。结论：乳腺癌组织低表达GATA3，GATA3表达与ER表达正相、与乳腺癌的发生、发展、转移和预后相关。
目的：探讨乳腺癌组织中GATA3的表达及与ER表达的关系，以及GATA3在乳腺癌发生中的意义。方法：采用免疫组化方法检测了上海市黄浦区中心医院2004年至2005年间外科手术切除的100例乳腺癌组织和61例癌旁乳腺组织中GATA3的表达水平，分析其与ER表达和乳腺癌患者临床病理特征间的关系。结果：GATA3在癌旁乳腺组织中的阳性表达率（82.0%）明显高于乳腺癌组织（60.0%）（χ2＝8.45，P<0.01），但在不同病理类型乳腺癌中表达无差异（χ2＝0.74，P>005）GATA3的表达和患者年龄、肿瘤大小无关（P>0.05），与乳腺癌组织分级、淋巴结转移相关（P<0.05）。乳腺癌组织中GATA3的阳性表达与ER的阳性表达存在着显著的正相关（r=0.49，P<0.01）。结论：乳腺癌组织低表达GATA3，GATA3表达与ER表达正相、与乳腺癌的发生、发展、转移和预后相关。
2011, 18(1):92-96.
DOI: 10.3872/j.issn.1007-385X.2011.1.020
Abstract:
神经轴突导向分子Netrin-1 通过与其受体结直肠癌缺陷基因（deleted in colorectal cancer, DCC）或UNC5同源物（UNC-5 Homolog, UNC5A-C, UNC5H1-3）家族成员结合传递信号，参与神经轴突的生长、定向迁移和神经元发育等生理功能。Netrin-1受体在没有配体结合的情况下，诱导细胞凋亡，故被归为“依赖性受体”（dependence receptor, DR）家族。Netrin-1受体在结直肠癌等多种肿瘤中下调或不表达，导致其表达下调的可能机制包括杂合性缺失、表观遗传学改变和转录调控等；过表达Netrin-1受体可抑制肿瘤细胞的增殖和迁移。Netrin-1及其受体家族是近年来鉴定出来的肿瘤参与分子，很可能是未来肿瘤治疗的新靶点。
神经轴突导向分子Netrin-1 通过与其受体结直肠癌缺陷基因（deleted in colorectal cancer, DCC）或UNC5同源物（UNC-5 Homolog, UNC5A-C, UNC5H1-3）家族成员结合传递信号，参与神经轴突的生长、定向迁移和神经元发育等生理功能。Netrin-1受体在没有配体结合的情况下，诱导细胞凋亡，故被归为“依赖性受体”（dependence receptor, DR）家族。Netrin-1受体在结直肠癌等多种肿瘤中下调或不表达，导致其表达下调的可能机制包括杂合性缺失、表观遗传学改变和转录调控等；过表达Netrin-1受体可抑制肿瘤细胞的增殖和迁移。Netrin-1及其受体家族是近年来鉴定出来的肿瘤参与分子，很可能是未来肿瘤治疗的新靶点。
2011, 18(1):97-100.
DOI: 10.3872/j.issn.1007-385X.2011.1.021
Abstract:
RNA结合蛋白（RNA-binding proteins，RBPs）是近年来发现的具有多种生物学作用的分子家族，其中，HuR属于胚胎致死异常视觉家族的RBPs，表达广泛。最初发现HuR与神经分化相关，通过与靶mRNA 3′UTR的ARE结合，在转录后水平调控RNA的稳定性和蛋白表达。近年发现，HuR与细胞增殖、肿瘤相关的炎症和血管生成密切相关，提示HuR可能参与了肿瘤发生、发展和转移过程。因此，以HuR为靶点的抗肿瘤策略可能为将来的肿瘤治疗带来希望。
RNA结合蛋白（RNA-binding proteins，RBPs）是近年来发现的具有多种生物学作用的分子家族，其中，HuR属于胚胎致死异常视觉家族的RBPs，表达广泛。最初发现HuR与神经分化相关，通过与靶mRNA 3′UTR的ARE结合，在转录后水平调控RNA的稳定性和蛋白表达。近年发现，HuR与细胞增殖、肿瘤相关的炎症和血管生成密切相关，提示HuR可能参与了肿瘤发生、发展和转移过程。因此，以HuR为靶点的抗肿瘤策略可能为将来的肿瘤治疗带来希望。
21 Effect of CMRF-35-like molecules on Toll-like receptor signal pathway and its mechanisms: A progress
2011, 18(1):101-104.
DOI: 10.3872/j.issn.1007-385X.2011.1.022
Abstract:
Toll样受体（Toll-like receptor, TLR）是人体天然免疫系统的主要参与者，TLR在免疫细胞中广泛表达，通过识别病原体相关分子模式（pathogen-associated molecule pattern, PAMP）来触发免疫应答，而对于TLR信号的调节机制的研究仍然是免疫学及其相关学科的热点领域。CMRF35样分子（CMRF-35-like molecules, CLMs）作为免疫细胞表面受体分子可能参与TLR信号的调节，其中CLM-1和CLM-8通过自身含有的免疫受体酪氨酸抑制基序触发对TLR信号的负向调控作用，而CLM-4通过与带有免疫受体酪氨酸活化基序的接头分子结合，促进TLR信号转导，调节免疫细胞对病原体的应答。对CLMs的深入研究将丰富人们对TLR信号调节分子机制的认识。
Toll样受体（Toll-like receptor, TLR）是人体天然免疫系统的主要参与者，TLR在免疫细胞中广泛表达，通过识别病原体相关分子模式（pathogen-associated molecule pattern, PAMP）来触发免疫应答，而对于TLR信号的调节机制的研究仍然是免疫学及其相关学科的热点领域。CMRF35样分子（CMRF-35-like molecules, CLMs）作为免疫细胞表面受体分子可能参与TLR信号的调节，其中CLM-1和CLM-8通过自身含有的免疫受体酪氨酸抑制基序触发对TLR信号的负向调控作用，而CLM-4通过与带有免疫受体酪氨酸活化基序的接头分子结合，促进TLR信号转导，调节免疫细胞对病原体的应答。对CLMs的深入研究将丰富人们对TLR信号调节分子机制的认识。
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
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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.