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Volume 14,Issue 1,2007 Table of Contents
2007, 14(1):2-6.
DOI: 10.3872/j.issn.1007-385X.2007.1.001
Abstract:
目的:观察5-氮-2′-脱氧胞苷(5-aza-2′-deoxycytidine,又称5-aza-CdR)对卵巢癌细胞SKOV3和3AO增殖凋亡及DNA错配基因hMLH1和hMLH2表达的影响。方法:以特异性甲基转移酶抑制剂5-aza-CdR0.5、5、50μmol/L处理人卵巢癌细胞SKOV3和3AO3d,继续常规培养7d后,采用MTT比色法观察细胞经药物处理前后的增殖活性,用流式细胞术分析5-aza-CdR对细胞凋亡影响,以半定量逆转录-聚合酶链反应(RT-PCR)检测细胞经5-aza-CdR处理前后DNA错配修复基因hMLH1和hMSH2mRNA表达水平的改变。结果:人卵巢癌细胞SKOV3和3AO经5-aza-CdR处理后,与对照组比较,0.5、5、50μmol/L均能明显抑制肿瘤细胞生长,随着5-aza-CdR浓度增加,细胞增殖速度下降。SKOV3经5-aza-CdR0.5、5、50μmol/L处理后细胞的凋亡率分别为(10.59±1.57)%、(17.52±1.72)%、(34.10±1.45)%,3A0经0.5、5、50μmol/L5-aza-CdR处理后细胞的凋亡率分别为(11.11±2.21)%、(17.24±1.11)%、(26.53±2.00)%,与对照组相比均有统计学意义(P<0.01);且凋亡率与剂量成正相关(FSKOV3=227.6,PSKOV3<0.01;F3AO=108.4,P3AO<0.01)。经5-aza-CdR处理后的两株卵巢癌细胞中hMLH1和hMLH2的mRNA表达量有不同程度的增加(P<0.01),且与药物存在剂量依赖性。结论:在人卵巢癌细胞株SKOV3和3AO中,5-aza-CdR可部分逆转hMLH1和hMLH2的失活,恢复其生长调控功能,抑制肿瘤细胞生长,并诱导细胞凋亡。
目的:观察5-氮-2′-脱氧胞苷(5-aza-2′-deoxycytidine,又称5-aza-CdR)对卵巢癌细胞SKOV3和3AO增殖凋亡及DNA错配基因hMLH1和hMLH2表达的影响。方法:以特异性甲基转移酶抑制剂5-aza-CdR0.5、5、50μmol/L处理人卵巢癌细胞SKOV3和3AO3d,继续常规培养7d后,采用MTT比色法观察细胞经药物处理前后的增殖活性,用流式细胞术分析5-aza-CdR对细胞凋亡影响,以半定量逆转录-聚合酶链反应(RT-PCR)检测细胞经5-aza-CdR处理前后DNA错配修复基因hMLH1和hMSH2mRNA表达水平的改变。结果:人卵巢癌细胞SKOV3和3AO经5-aza-CdR处理后,与对照组比较,0.5、5、50μmol/L均能明显抑制肿瘤细胞生长,随着5-aza-CdR浓度增加,细胞增殖速度下降。SKOV3经5-aza-CdR0.5、5、50μmol/L处理后细胞的凋亡率分别为(10.59±1.57)%、(17.52±1.72)%、(34.10±1.45)%,3A0经0.5、5、50μmol/L5-aza-CdR处理后细胞的凋亡率分别为(11.11±2.21)%、(17.24±1.11)%、(26.53±2.00)%,与对照组相比均有统计学意义(P<0.01);且凋亡率与剂量成正相关(FSKOV3=227.6,PSKOV3<0.01;F3AO=108.4,P3AO<0.01)。经5-aza-CdR处理后的两株卵巢癌细胞中hMLH1和hMLH2的mRNA表达量有不同程度的增加(P<0.01),且与药物存在剂量依赖性。结论:在人卵巢癌细胞株SKOV3和3AO中,5-aza-CdR可部分逆转hMLH1和hMLH2的失活,恢复其生长调控功能,抑制肿瘤细胞生长,并诱导细胞凋亡。
2007, 14(1):7-13.
DOI: 10.3872/j.issn.1007-385X.2007.1.002
Abstract:
放化疗导致的淋巴细胞减少，改变了免疫细胞亚群的格局，启动了免疫细胞的内源性增殖，重建了一个新的免疫微环境。其机制主要与抑制肿瘤免疫的调节性T细胞比例与功能显著下调；内源性细胞因子IL 7、IL 15等供应增多，淋巴细胞高效扩增；抗原提呈细胞功能增强和以非对称性为特点的淋巴细胞自稳性增生的改变等相关。经历了淋巴细胞减少状态下肿瘤免疫格局的改变后，机体消除了抑制肿瘤免疫的重要因素，打破了肿瘤免疫耐受。肿瘤放化疗后选择合适时间点与生物治疗联合治疗，可以诱导出优于单一生物治疗的抗肿瘤效应，部分修正了以往认为放化疗导致的淋巴细胞减少不利于抗肿瘤免疫生物治疗的观念，为临床攻克肿瘤开辟新的治疗途径。
放化疗导致的淋巴细胞减少，改变了免疫细胞亚群的格局，启动了免疫细胞的内源性增殖，重建了一个新的免疫微环境。其机制主要与抑制肿瘤免疫的调节性T细胞比例与功能显著下调；内源性细胞因子IL 7、IL 15等供应增多，淋巴细胞高效扩增；抗原提呈细胞功能增强和以非对称性为特点的淋巴细胞自稳性增生的改变等相关。经历了淋巴细胞减少状态下肿瘤免疫格局的改变后，机体消除了抑制肿瘤免疫的重要因素，打破了肿瘤免疫耐受。肿瘤放化疗后选择合适时间点与生物治疗联合治疗，可以诱导出优于单一生物治疗的抗肿瘤效应，部分修正了以往认为放化疗导致的淋巴细胞减少不利于抗肿瘤免疫生物治疗的观念，为临床攻克肿瘤开辟新的治疗途径。
Zhang Shenghai
,
Wu Jihong
,
Liu Xinjian
,
Chen Xiafang
,
Tian Yuhua
,
Xie Kuangcheng
,
Yu Hui
,
Pan Hong
,
Huang Qian
2007, 14(1):14-20.
DOI: 10.3872/j.issn.1007-385X.2007.1.003
Abstract:
To investigate the effects of chemotherapeutic agent Etoposide on transgene expression mediated by recombinant replication-defective adenovirus in tumor cell lines. Methods：Cultured tumor cells, including NCI-H446, NCI-H460, A549, SMMC-7721, SGC7901, SKBR-3, and BTT were infected by Ad-CMV-EGFP alone （MOI being 1 and 10）or in combination with Etoposide at different final concentrations (0.2, 2, 20, 40, 80, 100 and 200 μg/ml). GFP positive cell rates and the mean intensities of GFP fluorescence in tumor cells were detected by Fluorescence Activated Cell Sorting (FACS) after cultured with different strategies. EGFP protein expression in tumor cells was analyzed by Western blotting. Quantitative analysis of mRNA and DNA copies of EGFP in tumor cells were performed by RT-PCR and real-time PCR. Results: FACS results indicated that Etoposide efficiently enhanced the mean intensities of EGFP fluorescence to different degrees in all 7 cell lines but had no evident effects on the EGFP positive rate. Twenty-four hours after cultured at the presence of 10 MOI Ad5-CMV-EGFP and 40 μg/ml Etoposide, the mean intensities of EGFP fluorescence in NCI-H446, NCI-H460, A549, SMMC-7721, SGC7901, SKBR-3, and BTT cells were respectively 3.3, 3.5, 3.1, 6.2, 70, 5.4, and 3.4 folds that cultured with 10 MOI Ad5-CMV-EGFP alone. Western blotting showed that EGFP protein expression in cells co cultured with Ad5-CMV-EGFP and Etoposide was 2-5 times that cultured with 10 MOI Ad5-CMV-EGFP alone. EGFP mRNA expression had a similar tendency as EGFP protein, but the copies of EGFP DNA had no evident changes. Conclusion: Etoposide can enhance transgene expression mediated by recombination replication defective adenovirus in several tumor cell lines, which may play a role at the transcriptional level.
To investigate the effects of chemotherapeutic agent Etoposide on transgene expression mediated by recombinant replication-defective adenovirus in tumor cell lines. Methods：Cultured tumor cells, including NCI-H446, NCI-H460, A549, SMMC-7721, SGC7901, SKBR-3, and BTT were infected by Ad-CMV-EGFP alone （MOI being 1 and 10）or in combination with Etoposide at different final concentrations (0.2, 2, 20, 40, 80, 100 and 200 μg/ml). GFP positive cell rates and the mean intensities of GFP fluorescence in tumor cells were detected by Fluorescence Activated Cell Sorting (FACS) after cultured with different strategies. EGFP protein expression in tumor cells was analyzed by Western blotting. Quantitative analysis of mRNA and DNA copies of EGFP in tumor cells were performed by RT-PCR and real-time PCR. Results: FACS results indicated that Etoposide efficiently enhanced the mean intensities of EGFP fluorescence to different degrees in all 7 cell lines but had no evident effects on the EGFP positive rate. Twenty-four hours after cultured at the presence of 10 MOI Ad5-CMV-EGFP and 40 μg/ml Etoposide, the mean intensities of EGFP fluorescence in NCI-H446, NCI-H460, A549, SMMC-7721, SGC7901, SKBR-3, and BTT cells were respectively 3.3, 3.5, 3.1, 6.2, 70, 5.4, and 3.4 folds that cultured with 10 MOI Ad5-CMV-EGFP alone. Western blotting showed that EGFP protein expression in cells co cultured with Ad5-CMV-EGFP and Etoposide was 2-5 times that cultured with 10 MOI Ad5-CMV-EGFP alone. EGFP mRNA expression had a similar tendency as EGFP protein, but the copies of EGFP DNA had no evident changes. Conclusion: Etoposide can enhance transgene expression mediated by recombination replication defective adenovirus in several tumor cell lines, which may play a role at the transcriptional level.
ZHOU Bengen
,
XU Yanming
,
QIU Xiuchun
,
YANG Tongtao
,
JI Zhengang
,
GOU Yongsheng
,
REN Min
,
ZHOU Yong
,
YANG Angang
,
FAN Qingyu
2007, 14(1):21-25.
DOI: 10.3872/j.issn.1007-385X.2007.1.004
Abstract:
To investigate the proapoptotic effect of Her 2targeted recombinant caspase6 fusion protein on osteosarcoma SOSP9607 cells. Methods: Recombinant immunocasp6 was generated by fusing the genes of a signal peptide, a singlechain Her 2 antibody (e23sFv), a PEA translocation domain (PEA aa253364), and an active caspase6. The constructed immunocasp6 gene（e23sFvPEⅡcasp6） was cloned into pCMV plasmid and transfected into SOSP9607 cells. The target gene expression and the morphology of the transfected cells were observed by fluorescence and electron microscopy, and the proapoptotic effect of the recombinant gene was analyzed by Annexin VFITC staining, flow cytometry, and MTT assay. Results： The e23sFvPEⅡcasp6 fusion protein was detected in the cytoplasm of transfected SOSP9607 cells. The transfected cells presented the typical characteristics of apoptosis as detected by electron microscopy (cytoplasm concentration, chromatin condensation, vacuolation). Annexin VFITC staining revealed that the percentage of apoptotic cells in the transfectants of immunocasp6 genes was 31.4％,compared with 6.0％ in the control cells. MTT assay showed that the proliferation of immunocasp6 genetransfected SOSP9607 cells was much lower than that of non or mocktransfected cells. Conclusion： Her 2 targeted recombinant immunocasp6 fusion protein can express in SOSP9607 cells and induce apoptosis of SOSP9607 cells.
To investigate the proapoptotic effect of Her 2targeted recombinant caspase6 fusion protein on osteosarcoma SOSP9607 cells. Methods: Recombinant immunocasp6 was generated by fusing the genes of a signal peptide, a singlechain Her 2 antibody (e23sFv), a PEA translocation domain (PEA aa253364), and an active caspase6. The constructed immunocasp6 gene（e23sFvPEⅡcasp6） was cloned into pCMV plasmid and transfected into SOSP9607 cells. The target gene expression and the morphology of the transfected cells were observed by fluorescence and electron microscopy, and the proapoptotic effect of the recombinant gene was analyzed by Annexin VFITC staining, flow cytometry, and MTT assay. Results： The e23sFvPEⅡcasp6 fusion protein was detected in the cytoplasm of transfected SOSP9607 cells. The transfected cells presented the typical characteristics of apoptosis as detected by electron microscopy (cytoplasm concentration, chromatin condensation, vacuolation). Annexin VFITC staining revealed that the percentage of apoptotic cells in the transfectants of immunocasp6 genes was 31.4％,compared with 6.0％ in the control cells. MTT assay showed that the proliferation of immunocasp6 genetransfected SOSP9607 cells was much lower than that of non or mocktransfected cells. Conclusion： Her 2 targeted recombinant immunocasp6 fusion protein can express in SOSP9607 cells and induce apoptosis of SOSP9607 cells.
2007, 14(1):26-30.
DOI: 10.3872/j.issn.1007-385X.2007.1.005
Abstract:
To study the effects of natural killer cells with KIR(killer immunoglobulinlike receptor)/HLACw incompatibility on breast cancer cell lines in vitro, and to analyze the relationship between NKG2D (on NK cells) and MICA (on tumor cells) expression with NK cells alloreactivity against breast cancer.Methods: Monocytes were derived from 10 ml peripheral blood of healthy donors and patients of breast cancer by FicollHypaque. NK cells were purified by MASC separation system with NK Cell Isolation Kit. The polymerase chain reactionbased sequencespecific primer (PCRSSP) was used for KIR typing of NK cells and HLACw typing of tumor cells. NK cell cytotoxicity was assessed by MTT assays. Flow cytometry was used to investigate the expression level of NKG2D in NK cells and RTPCR was used to investigate the expression MICA in tumor cells. Results: The purity of enriched NK cells was more than 90% as determined by flow cytometry. The cytotoxicity of NK cells in KIR/HLACw compatible group was greatly higher than that of incompatible group. The cytotoxicity rates of NK cells of G1 and G2 group against MDAMB435s（G1 group）were (84.6±16.7)% and (25.4±8.9)%, respectively; against SKBr3（G1 group） were (70.2±11.8)% and (29.5±6.8)%, respectively; against MCF7（G2 group）were (33.1±5.7)% and (81.4±10.3)%; respectively. MICA expression was noticed in MCF7, MDAMB435s, and SKBr3 breast cancer cell lines. The expression of NKG2D on the NK cells was increased when cocultured with MCF7 cells. Conclusion: The cytotoxicity of NK cells against breast cancer cells is not mediated by KIR/HLACw incompatibility. MICA might increase the sensitivity of the breast cancer to NK cells by activating NKG2D on the NK cells.
To study the effects of natural killer cells with KIR(killer immunoglobulinlike receptor)/HLACw incompatibility on breast cancer cell lines in vitro, and to analyze the relationship between NKG2D (on NK cells) and MICA (on tumor cells) expression with NK cells alloreactivity against breast cancer.Methods: Monocytes were derived from 10 ml peripheral blood of healthy donors and patients of breast cancer by FicollHypaque. NK cells were purified by MASC separation system with NK Cell Isolation Kit. The polymerase chain reactionbased sequencespecific primer (PCRSSP) was used for KIR typing of NK cells and HLACw typing of tumor cells. NK cell cytotoxicity was assessed by MTT assays. Flow cytometry was used to investigate the expression level of NKG2D in NK cells and RTPCR was used to investigate the expression MICA in tumor cells. Results: The purity of enriched NK cells was more than 90% as determined by flow cytometry. The cytotoxicity of NK cells in KIR/HLACw compatible group was greatly higher than that of incompatible group. The cytotoxicity rates of NK cells of G1 and G2 group against MDAMB435s（G1 group）were (84.6±16.7)% and (25.4±8.9)%, respectively; against SKBr3（G1 group） were (70.2±11.8)% and (29.5±6.8)%, respectively; against MCF7（G2 group）were (33.1±5.7)% and (81.4±10.3)%; respectively. MICA expression was noticed in MCF7, MDAMB435s, and SKBr3 breast cancer cell lines. The expression of NKG2D on the NK cells was increased when cocultured with MCF7 cells. Conclusion: The cytotoxicity of NK cells against breast cancer cells is not mediated by KIR/HLACw incompatibility. MICA might increase the sensitivity of the breast cancer to NK cells by activating NKG2D on the NK cells.
2007, 14(1):31-36.
DOI: 10.3872/j.issn.1007-385X.2007.1.006
Abstract:
To obtain phagedisplayed ScFv library targeting tumor tissues and to screen for antibodies specifically binding to tumor vessels using in vivo phage display, so as to lay a foundation for diagnosis and treatment of cancer. Methods: The membrane proteins were extracted from the specimens of esophageal carcinoma, stomach carcinoma, brain cancer, lung cancer, and spinal cord tumor. The recombinant phageantibody system was used to construct a singlechain Fv fragment (ScFv) cDNA library from the total RNA of the BALB/c mice immunized with purified membrane protein. The specific primers of VH and VL were used to amplify the cDNA ofVH and VL, respectively, which were then assembled into ScFv gene with a specially constructed linker DNA. The ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated samples were transformed into competent E.coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage.Using the animal model of human cervical carcinoma (HeLa cells), sepecific phageScFvs were selected by phage displaying and panning in vivo. After four rounds, 24 phageScFvs, which were identified by PCR, were analyzed immunohistochemically. The ScFvs expressed in the tumor tissue slices and negative in control kidney tissue slices were sequenced. Results: Tomorsbearing animal models were established with 7 different kinds of carcinoma cell lines in BALB/c nude mice. It was found that inoculation with HeLa cells resulted in most satisfactory tumorigenesis in nude mice. A ScFv library of 1.6×106 was obtained and a tumor vessel specific phageScFv named ScFvH1 (VHlinkerVL) was selected from the library. Conclusion: A tumor targeting ScFv library has been successfully constructed and a tumor vesselspecifrc antibody has been identified from the library, which provides a new way for the early diagnosis and therapy of cancer.
To obtain phagedisplayed ScFv library targeting tumor tissues and to screen for antibodies specifically binding to tumor vessels using in vivo phage display, so as to lay a foundation for diagnosis and treatment of cancer. Methods: The membrane proteins were extracted from the specimens of esophageal carcinoma, stomach carcinoma, brain cancer, lung cancer, and spinal cord tumor. The recombinant phageantibody system was used to construct a singlechain Fv fragment (ScFv) cDNA library from the total RNA of the BALB/c mice immunized with purified membrane protein. The specific primers of VH and VL were used to amplify the cDNA ofVH and VL, respectively, which were then assembled into ScFv gene with a specially constructed linker DNA. The ScFv gene was ligated into the phagemid vector pCANTAB 5E and the ligated samples were transformed into competent E.coli TG1. The transformed cells were infected with M13KO7 helper phage to yield recombinant phage.Using the animal model of human cervical carcinoma (HeLa cells), sepecific phageScFvs were selected by phage displaying and panning in vivo. After four rounds, 24 phageScFvs, which were identified by PCR, were analyzed immunohistochemically. The ScFvs expressed in the tumor tissue slices and negative in control kidney tissue slices were sequenced. Results: Tomorsbearing animal models were established with 7 different kinds of carcinoma cell lines in BALB/c nude mice. It was found that inoculation with HeLa cells resulted in most satisfactory tumorigenesis in nude mice. A ScFv library of 1.6×106 was obtained and a tumor vessel specific phageScFv named ScFvH1 (VHlinkerVL) was selected from the library. Conclusion: A tumor targeting ScFv library has been successfully constructed and a tumor vesselspecifrc antibody has been identified from the library, which provides a new way for the early diagnosis and therapy of cancer.
2007, 14(1):37-41.
DOI: 10.3872/j.issn.1007-385X.2007.1.007
Abstract:
To investigate the influence of fetal liver AFT024 cells on the transfection efficiency of multidrug resistant gene 1 (MDR1) and the in vitro expansion of CD34+ cells derived from umbilical cord blood. Methods: CD34+ cells were isolated from human umbilical cord blood by MACS CD34 Progenitor Cell Isolation Kit and cocultured with AFT024 cells (AFT024 group) or cultured alone (control group) for 7 days. During the subsequent 14 days, retrovirus carrying MDR1 gene was supplemented twice a week to transfect CD34+ cells. On the 7th, 14th and 21st day after culture, the number of total nucleated cells (TNC) was counted, the ratio of CD34+ cells was assayed by flow cytometry (FCM) and the number of CD34+ cells was calculated, and colonyforming cells (CFC) were counted by methylcellulose cultures. RTPCR method was used to detect the level of MDR1 mRNA in the transfected cells. The expression and function of Pglycoprotein (Pgp) were evaluated by FCM assay and Rhodamine123 efflux assay, respectively. The gene transfection efficiency was calculated by drugresistant colonyforming cells assay. Results: (1) The MDR1 mRNA level in AFT024 group than that in control group. The gene transfection efficiency in AFT024 group was significantly higher than that in control group（46.0% vs 15.2%, P＜0.01)）. The expressions of Pgp in AFTO24 group and control group were (31.7±10.2)% and (12.6±3.9)%, respectively(P＜0.01). Pgp efflux functions in AFT024 group and control group were (35.5±11.4)% and (16.6±3.2)%, respectively (P＜0.01). (2) On the 7th day, the expansion folds of TNCs cells, CD34+ cells, and CFCs in control group were slightly higher than those in AFT024 group (P＞005). On the 14th day, the expansion fold of TNCs in control group was significantly higher than that in AFT024 group(P＜0.05), while the CD34+ cells in the AFT024 group were significantly more than those in control group(P＜005). There was no difference in the expansion folds of CFCs between the 2 groups. On the 21st day, the number of TNCs in AFT024 group was higher than those in control group(P＞0.05). The expansion folds of CD34+ cells and CFCs in the AFT024 group were significantly higher than that of the control group(P＜0.01). Conclusion: AFT024 cells can facilitate MDR1 gene transfection into CD34+ cells and improve the expansion of primitive hematopoietic cells in vitro.
To investigate the influence of fetal liver AFT024 cells on the transfection efficiency of multidrug resistant gene 1 (MDR1) and the in vitro expansion of CD34+ cells derived from umbilical cord blood. Methods: CD34+ cells were isolated from human umbilical cord blood by MACS CD34 Progenitor Cell Isolation Kit and cocultured with AFT024 cells (AFT024 group) or cultured alone (control group) for 7 days. During the subsequent 14 days, retrovirus carrying MDR1 gene was supplemented twice a week to transfect CD34+ cells. On the 7th, 14th and 21st day after culture, the number of total nucleated cells (TNC) was counted, the ratio of CD34+ cells was assayed by flow cytometry (FCM) and the number of CD34+ cells was calculated, and colonyforming cells (CFC) were counted by methylcellulose cultures. RTPCR method was used to detect the level of MDR1 mRNA in the transfected cells. The expression and function of Pglycoprotein (Pgp) were evaluated by FCM assay and Rhodamine123 efflux assay, respectively. The gene transfection efficiency was calculated by drugresistant colonyforming cells assay. Results: (1) The MDR1 mRNA level in AFT024 group than that in control group. The gene transfection efficiency in AFT024 group was significantly higher than that in control group（46.0% vs 15.2%, P＜0.01)）. The expressions of Pgp in AFTO24 group and control group were (31.7±10.2)% and (12.6±3.9)%, respectively(P＜0.01). Pgp efflux functions in AFT024 group and control group were (35.5±11.4)% and (16.6±3.2)%, respectively (P＜0.01). (2) On the 7th day, the expansion folds of TNCs cells, CD34+ cells, and CFCs in control group were slightly higher than those in AFT024 group (P＞005). On the 14th day, the expansion fold of TNCs in control group was significantly higher than that in AFT024 group(P＜0.05), while the CD34+ cells in the AFT024 group were significantly more than those in control group(P＜005). There was no difference in the expansion folds of CFCs between the 2 groups. On the 21st day, the number of TNCs in AFT024 group was higher than those in control group(P＞0.05). The expansion folds of CD34+ cells and CFCs in the AFT024 group were significantly higher than that of the control group(P＜0.01). Conclusion: AFT024 cells can facilitate MDR1 gene transfection into CD34+ cells and improve the expansion of primitive hematopoietic cells in vitro.
ZHENG Hongling
,
JIN Ningyi
,
LI Xiao
,
MI Zhiqiang
,
LIAN Hai
,
LI Chang
,
TIAN Mingyao
,
LI Xuemei
,
SUN Lili
2007, 14(1):42-46.
DOI: 10.3872/j.issn.1007-385X.2007.1.008
Abstract:
To investigate the combined antitumor effects of apoptin gene, hemagglutininneuramidinase gene (HN) of newcastle disease virus (NDV), and human IL18 gene(hIL18) on colon carcinoma cells HCT116in vitro. Methods:The recombinant plasmid pIRApoptinHNIL18 was introduced into human colon carcinoma cells HCT116 by liposomemediated transfection. The expression of the exogenous genes was detected by Western blotting and RTPCR. The antitumor effects of pIRApoptinHNIL18 on HCT116 cells were determined by AO/EB staining. The influence of pIRApoptinHNIL18 on mitochondrial transmembrane potential (△Ψ) and reactive oxygen species (ROS) were detected by flow cytometry (FCM). Sialic acid content was determined by using 3, 5dihydroxytoluene. Results: pIRApoptinHNIL18 transfection resulted in expression of the exogenous genes, repression of HCT116 cells, downregulation of △Ψ \[(94.41±8.17) % vs (30.70±8.01)%,P<005\] and Sialic acid content \[(0.33±0.06) mmol/L vs (0.09±003) mmol/L,P<0.01\], and upregulation of ROS \[(52.48±6.09)% vs (68.98±7.26)% P<0.05\]. Conclusion: pIRApoptinHNIL18 may inhibit the growth of HCT116 cells via mitochondrial pathwayinduced apoptosis.
To investigate the combined antitumor effects of apoptin gene, hemagglutininneuramidinase gene (HN) of newcastle disease virus (NDV), and human IL18 gene(hIL18) on colon carcinoma cells HCT116in vitro. Methods:The recombinant plasmid pIRApoptinHNIL18 was introduced into human colon carcinoma cells HCT116 by liposomemediated transfection. The expression of the exogenous genes was detected by Western blotting and RTPCR. The antitumor effects of pIRApoptinHNIL18 on HCT116 cells were determined by AO/EB staining. The influence of pIRApoptinHNIL18 on mitochondrial transmembrane potential (△Ψ) and reactive oxygen species (ROS) were detected by flow cytometry (FCM). Sialic acid content was determined by using 3, 5dihydroxytoluene. Results: pIRApoptinHNIL18 transfection resulted in expression of the exogenous genes, repression of HCT116 cells, downregulation of △Ψ \[(94.41±8.17) % vs (30.70±8.01)%,P<005\] and Sialic acid content \[(0.33±0.06) mmol/L vs (0.09±003) mmol/L,P<0.01\], and upregulation of ROS \[(52.48±6.09)% vs (68.98±7.26)% P<0.05\]. Conclusion: pIRApoptinHNIL18 may inhibit the growth of HCT116 cells via mitochondrial pathwayinduced apoptosis.
2007, 14(1):47-52.
DOI: 10.3872/j.issn.1007-385X.2007.1.009
Abstract:
To evaluate the inhibitory effect of Herceptin on HeLa, SiHa cells and to investigate the synergistic mechanism of Herceptin and Carboplatin. Methods: HeLa and SiHa cells were treated with Herceptin (at 5, 10，20, 40, and 80 μg/ml), Carboplatin (at 0.5, 1, 2, 4, and 8 μg/ml), and Her＋CBP\[（10＋1), (20＋2), and (40＋4) μg/ml\] separately. The untreated cells were taken as control. SP immunohistochemical method was used to detect the protein expression of HER2/neu and downstream Ras oncogene. MTT method was used to study the inhibition effect of Herceptin on HeLa, SiHa cells and its synergetic effect with carboplatin. FCM was used to detect the cell apoptosis and cell cycle. The mRNA expression of her2/neu and ras were assessed by RTPCR；the protein expression of HER2/neu and Ras were studied by Western blotting. Results: Herceptin significantly inhibited cervical cancer cells proliferation, and there was a synergistic effect when combined with carboplatin (P＜0.05, P＜0.01).Herceptin induced cell apoptosis and arrested cell at G1 phase. When combined with Carboplatin, Herceptin induced more severe apoptosis and arrested cell at G2 phase; meanwhile, the cells of S and also decreased. The mRNA and protein expressions of HER2/neuand Ras were decreased after treated with Herceptin alone and in combination with Carboplatin (P＜0.05). The inhibitory effect of Herceptin and Carboplatin was more obvious on HeLa cells. Conclusion: Herceptin alone or in combination with Carboplatin can greatly inhibit the growth to HeLa and SiHa cells. The synergistic mechanism of Herceptin with Carboplatin is related to the double blockage of cell cycle. The inhibitory effect is more obvious on HeLa cells. Herceptin inhibits proliferation of cervical cancer cells by restraining Ras/MAPK pathway.
To evaluate the inhibitory effect of Herceptin on HeLa, SiHa cells and to investigate the synergistic mechanism of Herceptin and Carboplatin. Methods: HeLa and SiHa cells were treated with Herceptin (at 5, 10，20, 40, and 80 μg/ml), Carboplatin (at 0.5, 1, 2, 4, and 8 μg/ml), and Her＋CBP\[（10＋1), (20＋2), and (40＋4) μg/ml\] separately. The untreated cells were taken as control. SP immunohistochemical method was used to detect the protein expression of HER2/neu and downstream Ras oncogene. MTT method was used to study the inhibition effect of Herceptin on HeLa, SiHa cells and its synergetic effect with carboplatin. FCM was used to detect the cell apoptosis and cell cycle. The mRNA expression of her2/neu and ras were assessed by RTPCR；the protein expression of HER2/neu and Ras were studied by Western blotting. Results: Herceptin significantly inhibited cervical cancer cells proliferation, and there was a synergistic effect when combined with carboplatin (P＜0.05, P＜0.01).Herceptin induced cell apoptosis and arrested cell at G1 phase. When combined with Carboplatin, Herceptin induced more severe apoptosis and arrested cell at G2 phase; meanwhile, the cells of S and also decreased. The mRNA and protein expressions of HER2/neuand Ras were decreased after treated with Herceptin alone and in combination with Carboplatin (P＜0.05). The inhibitory effect of Herceptin and Carboplatin was more obvious on HeLa cells. Conclusion: Herceptin alone or in combination with Carboplatin can greatly inhibit the growth to HeLa and SiHa cells. The synergistic mechanism of Herceptin with Carboplatin is related to the double blockage of cell cycle. The inhibitory effect is more obvious on HeLa cells. Herceptin inhibits proliferation of cervical cancer cells by restraining Ras/MAPK pathway.
CHU Xiaoyuan
,
WANG Jiejun
,
CHEN Longbang
,
WANG Jinghua
,
GUAN Xiaoxiang
,
GENG Huaicheng
,
ZHANG Qun
,
SONG Haizhu
,
WANG Hao
2007, 14(1):53-58.
DOI: 10.3872/j.issn.1007-385X.2007.1.010
Abstract:
To investigate the specific antitumor effect of mIL12 gene modified dendritic cells (DC) transfected with the total RNA of CT26 murine colon carcinoma. Methods: DC were cultured from murine bone marrow in the presence of rmGMCSF and rmIL4.The purity of DC was detected by flowcytometry. Adenovirus carrying mIL12 gene was proliferated in 293 cells and was transfected into DC. The total RNA of CT26 was extracted by Trizol reagent and was introduced into mIL12 gene modified DC by TransMessenger in vitro. The modified DC were used to immunize mice. The in vitro and in vivo levels of mIL12 and the in vivo activity of cytotoxic T lymphocyte（CTL）were examined by ELISA assay and modified LDH release assay, respectively. The tumor growth and animal survival time of immunized mice were estimated after they were challenged with parental tumor cells. Results: DC were successfully obtained from the culture of murine bone marrow, with CD11c+ accounting for over 90%. Vaccination with mIL12 gene modified DC transfected with the total RNA of CT26 induced strong specific CTL activity in vitro. All the immunized mice survived for a long term. Vaccination with AdLacZ modified DC and DC transfected with the total RNA induced moderate specific CTL activity in vitro（P<0.01）, and 60% of the immunized mice survived for a long time. There was no specific CTL activity in the mice immunized with DC and PBS and all the mice died. Conclusion: mIL12 gene modified DC transfected with the total RNA of CT26 can effectively present endogenetic tumor antigen and induce strong CTL activity, thus inducing more specific antitumor immune response.
To investigate the specific antitumor effect of mIL12 gene modified dendritic cells (DC) transfected with the total RNA of CT26 murine colon carcinoma. Methods: DC were cultured from murine bone marrow in the presence of rmGMCSF and rmIL4.The purity of DC was detected by flowcytometry. Adenovirus carrying mIL12 gene was proliferated in 293 cells and was transfected into DC. The total RNA of CT26 was extracted by Trizol reagent and was introduced into mIL12 gene modified DC by TransMessenger in vitro. The modified DC were used to immunize mice. The in vitro and in vivo levels of mIL12 and the in vivo activity of cytotoxic T lymphocyte（CTL）were examined by ELISA assay and modified LDH release assay, respectively. The tumor growth and animal survival time of immunized mice were estimated after they were challenged with parental tumor cells. Results: DC were successfully obtained from the culture of murine bone marrow, with CD11c+ accounting for over 90%. Vaccination with mIL12 gene modified DC transfected with the total RNA of CT26 induced strong specific CTL activity in vitro. All the immunized mice survived for a long term. Vaccination with AdLacZ modified DC and DC transfected with the total RNA induced moderate specific CTL activity in vitro（P<0.01）, and 60% of the immunized mice survived for a long time. There was no specific CTL activity in the mice immunized with DC and PBS and all the mice died. Conclusion: mIL12 gene modified DC transfected with the total RNA of CT26 can effectively present endogenetic tumor antigen and induce strong CTL activity, thus inducing more specific antitumor immune response.
11 Enhancement of cancer cell radiosensitivity by adenovirus vector carrying siRNA of Survivin gene
2007, 14(1):59-63.
DOI: 10.3872/j.issn.1007-385X.2007.1.011
Abstract:
To construct a recombinant adenoviral vector carrying the specific siRNA of Survivin gene, and to observe its effect on the expression of Survivin gene and on the radiosensitivity of cancer cells in tumorbearing mice. Methods: The specific siRNA of Survivin gene was designed and synthesized, and a recombinant adenovirus AdEGFPsiRNA was subsequently constructed. SMMC 7721 xenograft models were established with nude mice and were divided into the following 5 groups: siRNA+radiotherapy and siRNA groups (intratumoral injection of AdEGFPsiRNA), siRNA(-) group (injected every other day with AdEGFPsiRNA\[-\], 2×108 pfu/100 μl per time, total 5 times), pure radiotherapy group and blank control groups (injected with the same volume of normal saline). On day 10, 12, 14, and 16, the mice in siRNA+radiotherapy and pure radiotherapy groups were given 5 Gy/time radiotherapy. The tumor volumes were measured regularly. The expression of Survivin in tumor tissues was determined immunohistochemically. Results: Adenovirus AdEGFPsiRNA harboring the specific siRNA of Survivin gene and enhanced green fluorescent protein gene (EGFP) was successfully recombined. The growth of SMMC 7721 xenografts in nude mice was inhibited after injecting AdEGFPsiRNA, with the inhibition rate being 56.2%. The inhibition rate in AdEGFPsiRNA therapy + radiotherapy increased to 82.6%. Immunohistochemistry study showed that the specific siRNA markedly silenced the expression of Survivin gene in hepatocarcinoma cells. Conclusion: The specific siRNA can markedly silence Survivin gene and subsequently inhibit the growth of cancer; meanwhile, it can also increase the radiosensitivity of cancer cells so as to improve the treatment effect.
To construct a recombinant adenoviral vector carrying the specific siRNA of Survivin gene, and to observe its effect on the expression of Survivin gene and on the radiosensitivity of cancer cells in tumorbearing mice. Methods: The specific siRNA of Survivin gene was designed and synthesized, and a recombinant adenovirus AdEGFPsiRNA was subsequently constructed. SMMC 7721 xenograft models were established with nude mice and were divided into the following 5 groups: siRNA+radiotherapy and siRNA groups (intratumoral injection of AdEGFPsiRNA), siRNA(-) group (injected every other day with AdEGFPsiRNA\[-\], 2×108 pfu/100 μl per time, total 5 times), pure radiotherapy group and blank control groups (injected with the same volume of normal saline). On day 10, 12, 14, and 16, the mice in siRNA+radiotherapy and pure radiotherapy groups were given 5 Gy/time radiotherapy. The tumor volumes were measured regularly. The expression of Survivin in tumor tissues was determined immunohistochemically. Results: Adenovirus AdEGFPsiRNA harboring the specific siRNA of Survivin gene and enhanced green fluorescent protein gene (EGFP) was successfully recombined. The growth of SMMC 7721 xenografts in nude mice was inhibited after injecting AdEGFPsiRNA, with the inhibition rate being 56.2%. The inhibition rate in AdEGFPsiRNA therapy + radiotherapy increased to 82.6%. Immunohistochemistry study showed that the specific siRNA markedly silenced the expression of Survivin gene in hepatocarcinoma cells. Conclusion: The specific siRNA can markedly silence Survivin gene and subsequently inhibit the growth of cancer; meanwhile, it can also increase the radiosensitivity of cancer cells so as to improve the treatment effect.
2007, 14(1):64-68.
DOI: 10.3872/j.issn.1007-385X.2007.1.012
Abstract:
To observe the effect of 5aza2′deoxycytidine on proliferation and apoptosis of ovarian cancer cell lines (SKOV3 and 3AO) and on expression of mismatch repair (MMR) genes (hMLH1 and hMSH2) in SKOV3 and 3AO cells. Methods:Human ovarian cancer cell lines SKOV3 and 3AO were treated for 3 d with 5azaCdR (0.5, 5, 50 μmol/L), a specific demethylation agent, and then cultured in RPMI 1640 medium for another 7 d. The cells growth was observed by MTT assay before and after 5azaCdR treatment and the cell apoptosis was analyzed by flow cytometry. The expression of hMLH1 and hMSH2 mRNA was examined by semiquantitative reverse transcription polymerase chain reaction (RTPCR). Results: 5azaCdR (0.5, 5, 50 μmol/L) obviously inhibited the growth of SKOV3 and 3AO cells compared in a concentration dependent manner. The apoptosis rates of SKOV3 cells were (10.59±1.57)%, (17.52±172)%, (34.10±1.45)% after treated with 0.5, 5, 50 μmol/L 5azaCdR, respectively; and the apoptosis rates of 3AO cells were (11.11±2.21)%, (17.24±1.11)%, and (26.53±2.00)%, respectively, which were all markedly higher than those of control group（P <0.01）. We also found that the apoptosis rate was positively correlated with 5azaCdR concentration（FSKOV3=227.6，PSKOV3<0.01； F3AO=108.4，P3AO<0.01）. 5azaCdR treatment also increased the expression levels of hMLH1 and hMSH2 mRNA in a concentrationdependent manner (P <0.01). Conclusion: In human ovarian cancer cell line SKOV3 and 3AO，5azaCdR can partially reverse the deactivation of hMLH1 and hMSH2 of MMR genes and help hMLH1 and hMSH2 regain their function of growth regulation, thus inhibit the proliferation of tumor cells and induce cell apoptosis.
To observe the effect of 5aza2′deoxycytidine on proliferation and apoptosis of ovarian cancer cell lines (SKOV3 and 3AO) and on expression of mismatch repair (MMR) genes (hMLH1 and hMSH2) in SKOV3 and 3AO cells. Methods:Human ovarian cancer cell lines SKOV3 and 3AO were treated for 3 d with 5azaCdR (0.5, 5, 50 μmol/L), a specific demethylation agent, and then cultured in RPMI 1640 medium for another 7 d. The cells growth was observed by MTT assay before and after 5azaCdR treatment and the cell apoptosis was analyzed by flow cytometry. The expression of hMLH1 and hMSH2 mRNA was examined by semiquantitative reverse transcription polymerase chain reaction (RTPCR). Results: 5azaCdR (0.5, 5, 50 μmol/L) obviously inhibited the growth of SKOV3 and 3AO cells compared in a concentration dependent manner. The apoptosis rates of SKOV3 cells were (10.59±1.57)%, (17.52±172)%, (34.10±1.45)% after treated with 0.5, 5, 50 μmol/L 5azaCdR, respectively; and the apoptosis rates of 3AO cells were (11.11±2.21)%, (17.24±1.11)%, and (26.53±2.00)%, respectively, which were all markedly higher than those of control group（P <0.01）. We also found that the apoptosis rate was positively correlated with 5azaCdR concentration（FSKOV3=227.6，PSKOV3<0.01； F3AO=108.4，P3AO<0.01）. 5azaCdR treatment also increased the expression levels of hMLH1 and hMSH2 mRNA in a concentrationdependent manner (P <0.01). Conclusion: In human ovarian cancer cell line SKOV3 and 3AO，5azaCdR can partially reverse the deactivation of hMLH1 and hMSH2 of MMR genes and help hMLH1 and hMSH2 regain their function of growth regulation, thus inhibit the proliferation of tumor cells and induce cell apoptosis.
2007, 14(1):69-74.
DOI: 10.3872/j.issn.1007-385X.2007.1.013
Abstract:
To investigate the expression of somatostatin receptors (SSTR) subtypes (SSTTR2A, and SSTR5) and epidermal growth factor receptor (EGFR) in nonsmall cell lung cancer (NSCLC) and their clinical relevance. Methods: The expressions of SSTR2A, SSTR5, and EGFR in 62 NSCLC specimens and 7 adjacent normal lung tissues were examined using immunohistochemical method (SP). All patients were followed up in this study. Results: The positive rates of SSTR2A and SSTR5 in the 62 specimens were 48.3% (30 cases) and 70.9% (44 cases), respectively. The positive rates of SSTR2A and SSTR5 were closely related to TNM stage (P<0.05), but not to patients' ages, sexes, smoking history, pathological types, tumor sizes, and lymph metastases (P>0.05). In the same group, EGFR expressed in 56.4% (35 cases) of NSCLC specimens, but was not expressed in the 7 normal spesimens. The positive rate of EGFR was not related to the ages, sexes, smoking history, histological types, tumor sizes, TNM stages, pathological classification, and lymph metastases (P>0.05). There was a negative correlation between the EGFR expression with the expression of SSTR2A and SSTR5 in NSCLC tissues. The 3year survival rates were 64.5% and 65.9% in patients positive of SSTR5 and SSTR2A, respectively; and were 45.2% and 22.2% for those negative of SSTR5 and SSTR2A, respectively (P<0.05). The 3year survival rate was 30.8% for patients positive of EGFR protein and 69.4% for those negative of it (P<0.05). Conclusion: The expressions of SSTR2A, SSTR5, and EGFR can reflect the biology behavior of lung cancer, and the examination of them may be helpful for evaluation of the lymph node metastases, pathological classification, and prognosis of NSCLC.
To investigate the expression of somatostatin receptors (SSTR) subtypes (SSTTR2A, and SSTR5) and epidermal growth factor receptor (EGFR) in nonsmall cell lung cancer (NSCLC) and their clinical relevance. Methods: The expressions of SSTR2A, SSTR5, and EGFR in 62 NSCLC specimens and 7 adjacent normal lung tissues were examined using immunohistochemical method (SP). All patients were followed up in this study. Results: The positive rates of SSTR2A and SSTR5 in the 62 specimens were 48.3% (30 cases) and 70.9% (44 cases), respectively. The positive rates of SSTR2A and SSTR5 were closely related to TNM stage (P<0.05), but not to patients' ages, sexes, smoking history, pathological types, tumor sizes, and lymph metastases (P>0.05). In the same group, EGFR expressed in 56.4% (35 cases) of NSCLC specimens, but was not expressed in the 7 normal spesimens. The positive rate of EGFR was not related to the ages, sexes, smoking history, histological types, tumor sizes, TNM stages, pathological classification, and lymph metastases (P>0.05). There was a negative correlation between the EGFR expression with the expression of SSTR2A and SSTR5 in NSCLC tissues. The 3year survival rates were 64.5% and 65.9% in patients positive of SSTR5 and SSTR2A, respectively; and were 45.2% and 22.2% for those negative of SSTR5 and SSTR2A, respectively (P<0.05). The 3year survival rate was 30.8% for patients positive of EGFR protein and 69.4% for those negative of it (P<0.05). Conclusion: The expressions of SSTR2A, SSTR5, and EGFR can reflect the biology behavior of lung cancer, and the examination of them may be helpful for evaluation of the lymph node metastases, pathological classification, and prognosis of NSCLC.
2007, 14(1):75-78.
DOI: 10.3872/j.issn.1007-385X.2007.1.014
Abstract:
To construct a replicationdeficient adenovirus carrying p16 gene and to investigate its antitumor activity on human gastric cancer xenografts in nude mice. Methods: p16 cDNA was amplified by PCR and was inserted into the plasmid pSuCMV, the latter was then used to recombine the replicationdeficient adenovirus AdCMVp16 in 293 cells. Human SGC7901 gastric cancer xenograft models were established in nude mice and were divided into 3 groups: AdCMVp16, AdLacZ, and control groups. Mice in AdCMVp16 group received intratumoral injections of 2×10 8 pfu/100 l AdCMVp16 ( injected every other day for 5 times). Mice in control group received the same volume of virus preserving solution. The tumor volumes were measured at predefined time points. The antitumor effect of AdCMVp16 was observed by p16 immunochemical study and TUNEL detection of cell apoptosis. Results:The replicationdeficient adenovirus expressing p16 gene evidently inhibited the growth of human gastric cancer xenografts in nude mice (P<0.01), with the inhibition rate being 58.12%; but AdLacZ showed no obvious effect on the growth of xenografts (P>0.05), only with a inhibition rate of 4.26%. The pathological examination showed that apoptoses were the main changes in AdCMVp16 group, and p16 gene was found in the cancer cells. Conclusion: The replicationdeficient adenovirus harboring p16 gene can recover the expression of p16 in gastric cancer cells and subsequently inhibit the growth of human gastric cancer.
To construct a replicationdeficient adenovirus carrying p16 gene and to investigate its antitumor activity on human gastric cancer xenografts in nude mice. Methods: p16 cDNA was amplified by PCR and was inserted into the plasmid pSuCMV, the latter was then used to recombine the replicationdeficient adenovirus AdCMVp16 in 293 cells. Human SGC7901 gastric cancer xenograft models were established in nude mice and were divided into 3 groups: AdCMVp16, AdLacZ, and control groups. Mice in AdCMVp16 group received intratumoral injections of 2×10 8 pfu/100 l AdCMVp16 ( injected every other day for 5 times). Mice in control group received the same volume of virus preserving solution. The tumor volumes were measured at predefined time points. The antitumor effect of AdCMVp16 was observed by p16 immunochemical study and TUNEL detection of cell apoptosis. Results:The replicationdeficient adenovirus expressing p16 gene evidently inhibited the growth of human gastric cancer xenografts in nude mice (P<0.01), with the inhibition rate being 58.12%; but AdLacZ showed no obvious effect on the growth of xenografts (P>0.05), only with a inhibition rate of 4.26%. The pathological examination showed that apoptoses were the main changes in AdCMVp16 group, and p16 gene was found in the cancer cells. Conclusion: The replicationdeficient adenovirus harboring p16 gene can recover the expression of p16 in gastric cancer cells and subsequently inhibit the growth of human gastric cancer.
2007, 14(1):79-80.
DOI: 10.3872/j.issn.1007-385X.2007.1.015
Abstract:
检测胃癌和癌前病变组织中的端粒酶基因的表达，探讨其与胃癌发生的关系和在胃癌诊断中的意义。方法 : 收集本院病理科不同类型胃癌96例，胃黏膜轻、中、重度异型增生57例，肠上皮化生19例组织存档蜡块。用原位杂交法检测端粒酶基因（human telomerase RNA,hTR）和端粒酶催化亚单位基因（human telomerase reversetranscriptase, hTRT）在胃癌、胃黏膜异型增生和肠上皮化生组织中的表达。 结果 胃癌、异型增生和肠上皮化生组织中hTR和hTRT的阳性率分别为86.5%和88.5%、36.8%和36.8%、5.2%和5.2%。胃癌组织端粒酶基因表达明显高于癌前病变组织。结论: 端粒酶可能参与了胃癌的发生与发展，且与肿瘤细胞的分化程度和生物学行为有关。
检测胃癌和癌前病变组织中的端粒酶基因的表达，探讨其与胃癌发生的关系和在胃癌诊断中的意义。方法 : 收集本院病理科不同类型胃癌96例，胃黏膜轻、中、重度异型增生57例，肠上皮化生19例组织存档蜡块。用原位杂交法检测端粒酶基因（human telomerase RNA,hTR）和端粒酶催化亚单位基因（human telomerase reversetranscriptase, hTRT）在胃癌、胃黏膜异型增生和肠上皮化生组织中的表达。 结果 胃癌、异型增生和肠上皮化生组织中hTR和hTRT的阳性率分别为86.5%和88.5%、36.8%和36.8%、5.2%和5.2%。胃癌组织端粒酶基因表达明显高于癌前病变组织。结论: 端粒酶可能参与了胃癌的发生与发展，且与肿瘤细胞的分化程度和生物学行为有关。
2007, 14(1):81-82.
DOI: 10.3872/j.issn.1007-385X.2007.1.016
Abstract:
Objective:To investigate the specific anti-tumor effect of mIL-12 gene modified dendritic cells(DC)transfected with the total RNA of CT-26 murine colon carcinoma.Methods:DC were cultured from murine bone marrow in the presence of rmGM-CSF and rmIL-4.The p
Objective:To investigate the specific anti-tumor effect of mIL-12 gene modified dendritic cells(DC)transfected with the total RNA of CT-26 murine colon carcinoma.Methods:DC were cultured from murine bone marrow in the presence of rmGM-CSF and rmIL-4.The p
2007, 14(1):83-86.
DOI: 10.3872/j.issn.1007-385X.2007.1.017
Abstract:
昼夜节律钟基因(circadian clock genes)在分子水平构成生物钟，使得生物体的细胞分裂、免疫和内分泌等各项生命活动呈现周期性、有序性和协同性。昼夜节律钟基因控制着细胞周期，昼夜节律紊乱会增加癌症发病率；时辰化疗较之常规化疗能够进一步提高治疗指数，减少不良反应，改善患者生活质量；昼夜节律是肿瘤患者重要的生存预后指标；抗肿瘤免疫调节制剂给药也需遵循时间规律。昼夜节律钟基因与细胞周期关系的进一步阐明可能带来对于癌症成因、播散和转移机制认识的深化，并可能带来择时诊疗的新模式。
昼夜节律钟基因(circadian clock genes)在分子水平构成生物钟，使得生物体的细胞分裂、免疫和内分泌等各项生命活动呈现周期性、有序性和协同性。昼夜节律钟基因控制着细胞周期，昼夜节律紊乱会增加癌症发病率；时辰化疗较之常规化疗能够进一步提高治疗指数，减少不良反应，改善患者生活质量；昼夜节律是肿瘤患者重要的生存预后指标；抗肿瘤免疫调节制剂给药也需遵循时间规律。昼夜节律钟基因与细胞周期关系的进一步阐明可能带来对于癌症成因、播散和转移机制认识的深化，并可能带来择时诊疗的新模式。
2007, 14(1):87-89.
DOI: 10.3872/j.issn.1007-385X.2007.1.018
Abstract:
移植物抗宿主病（graftversushost disease,GVHD）是异基因造血干细胞移植(allogeneic haematopoietic stem cell transplantation,AlloHSCT)的主要并发症，主要由供者T淋巴细胞介导，如何有效控制GVHD，从而提高移植患者的生存率和生活质量一直是研究热点。记忆CD8+ T细胞来源于效应CD8+T细胞，至少分为两个亚群：中央型记忆T细胞和效应型记忆T细胞，两者在功能上和游走性方面均有不同。记忆CD8+ T细胞依赖IL2、IL7和IL15等细胞因子支持而稳定生存，当再次遇到能迅速地增生和分化为效应细胞并参与二次免疫应答，在骨髓移植后GVHD的维持中起着重要作用。记忆性干细胞表达CD44LoCD62Lhi，具有自我复制更新的能力，在受到刺激时可产生效应细胞和所有记忆性细胞亚群。供、受者抗原提呈细胞对异体反应性记忆T细胞的发生和调节起不同的作用。
移植物抗宿主病（graftversushost disease,GVHD）是异基因造血干细胞移植(allogeneic haematopoietic stem cell transplantation,AlloHSCT)的主要并发症，主要由供者T淋巴细胞介导，如何有效控制GVHD，从而提高移植患者的生存率和生活质量一直是研究热点。记忆CD8+ T细胞来源于效应CD8+T细胞，至少分为两个亚群：中央型记忆T细胞和效应型记忆T细胞，两者在功能上和游走性方面均有不同。记忆CD8+ T细胞依赖IL2、IL7和IL15等细胞因子支持而稳定生存，当再次遇到能迅速地增生和分化为效应细胞并参与二次免疫应答，在骨髓移植后GVHD的维持中起着重要作用。记忆性干细胞表达CD44LoCD62Lhi，具有自我复制更新的能力，在受到刺激时可产生效应细胞和所有记忆性细胞亚群。供、受者抗原提呈细胞对异体反应性记忆T细胞的发生和调节起不同的作用。
2007, 14(1):90-93.
DOI: 10.3872/j.issn.1007-385X.2007.1.019
Abstract:
传统观念认为化疗以后机体免疫功能低下，化疗后不应随即实施肿瘤免疫治疗。由于缺乏客观、精确的指标从体内实验去评判化疗与免疫治疗的关系，这方面的研究工作一直寥寥无几。近年来，随着肿瘤学和免疫学研究的深入发展，上述课题的探索取得了一些欣喜的结果，即化疗药物诱导的凋亡肿瘤细胞能够有效诱发免疫应答，并通过增加肿瘤抗原交叉提呈量、减少Treg细胞的数量并抑制其功能、促进肿瘤细胞减灭、使APCs对CD40信号的敏感性增强、影响机体细胞因子网络、影响效应T细胞移行及肿瘤免疫记忆产生等途径，对随后的免疫治疗起到增益作用。该增益作用已被日益增多的动物实验和临床报告所证实。
传统观念认为化疗以后机体免疫功能低下，化疗后不应随即实施肿瘤免疫治疗。由于缺乏客观、精确的指标从体内实验去评判化疗与免疫治疗的关系，这方面的研究工作一直寥寥无几。近年来，随着肿瘤学和免疫学研究的深入发展，上述课题的探索取得了一些欣喜的结果，即化疗药物诱导的凋亡肿瘤细胞能够有效诱发免疫应答，并通过增加肿瘤抗原交叉提呈量、减少Treg细胞的数量并抑制其功能、促进肿瘤细胞减灭、使APCs对CD40信号的敏感性增强、影响机体细胞因子网络、影响效应T细胞移行及肿瘤免疫记忆产生等途径，对随后的免疫治疗起到增益作用。该增益作用已被日益增多的动物实验和临床报告所证实。
2007, 14(1):94-96.
DOI: 10.3872/j.issn.1007-385X.2007.1.020
Abstract:
恶性肿瘤的发生常常是多个基因改变的结果，单一基因治疗的抗肿瘤效应往往十分有限，联合应用不同特性的细胞因子或趋化因子抗肿瘤是近年来的研究热点。本文着重综述趋化因子和B7分子在肿瘤基因治疗中的联合应用。B7分子是重要的共刺激分子，可为T细胞激活提供第二信号；趋化因子可通过趋化免疫活性细胞，部分趋化因子还可抑制肿瘤血管生成，从而产生抗肿瘤效应。共转染趋化因子和B7分子除通过“招引活化”的机制外，还可抑制CD4+CD25+调节性T细胞亚型向肿瘤局部浸润，进而改善肿瘤局部免疫抑制状态；同时可抑制肿瘤血管生成，从而通过多个不同的途径，产生更强的抗肿瘤效应。但趋化因子具有抗肿瘤和促肿瘤双向性，因此选择合适的趋化因子与B7分子联合应用是至关重要的。
恶性肿瘤的发生常常是多个基因改变的结果，单一基因治疗的抗肿瘤效应往往十分有限，联合应用不同特性的细胞因子或趋化因子抗肿瘤是近年来的研究热点。本文着重综述趋化因子和B7分子在肿瘤基因治疗中的联合应用。B7分子是重要的共刺激分子，可为T细胞激活提供第二信号；趋化因子可通过趋化免疫活性细胞，部分趋化因子还可抑制肿瘤血管生成，从而产生抗肿瘤效应。共转染趋化因子和B7分子除通过“招引活化”的机制外，还可抑制CD4+CD25+调节性T细胞亚型向肿瘤局部浸润，进而改善肿瘤局部免疫抑制状态；同时可抑制肿瘤血管生成，从而通过多个不同的途径，产生更强的抗肿瘤效应。但趋化因子具有抗肿瘤和促肿瘤双向性，因此选择合适的趋化因子与B7分子联合应用是至关重要的。
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- 《中国肿瘤生物治疗杂志》
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Supported by:Beijing E-Tiller Technology Development Co., Ltd.