高迁移率族蛋白1对奥沙利铂诱导的SGC-7901胃癌细胞凋亡的影响  

Role of high mobility group box 1 in oxaliplatin-inducedapoptosis in SGC-7901 gastric cancer cells

作　　者：沈磊[1] 胡梅琮[1] 吴亚欧[1] 张梅[1] 李群[1] 桂丽[1] 胡春松[1] 张林杰[1] 黄保军[1] 

机构地区：[1]安徽医科大学免疫学教研室,合肥230032

出　　处：《安徽医科大学学报》2011年第6期527-531,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:30972791)

摘　　要：目的构建pDsRED2-HMGB1重组质粒,探讨转染高迁移率族蛋白1(HMGB1)基因对奥沙利铂诱导的SGC-7901细胞凋亡的影响。方法用逆转录聚合酶链反应(RT-PCR)扩增HMGB1 mRNA全编码序列,将PCR产物插入到pD-sRED2-N1载体的Xho I和EcoR I位点,构建pDsRED2-HMGB1重组质粒;通过脂质体法转染SGC-7901胃癌细胞,荧光显微镜检测HMGB1红色荧光融合蛋白表达。Westernblot鉴定HMGB1蛋白表达。应用流式细胞仪(Annexin-V/PI标记)分析HMGB1过表达对奥沙利铂诱导胃癌细胞凋亡的影响。结果成功构建真核表达质粒pDsRED2-HMGB1,转染SGC-7901细胞后,荧光显微镜下可见细胞内有HMGB1红色荧光融合蛋白的表达;流式细胞术检测发现转染pD-sRED2-HMGB1后SGC-7901细胞凋亡水平较转染pDsRED2-N1组下降22.4%。结论 HMGB1过表达可抑制奥沙利铂诱导的SGC-7901胃癌细胞凋亡。Objective To construct the eukaryotic expression vector pDsRED2-HMGB1, and to explore the role ofHMGB1 in oxaliplatin-induced apoptosis in SGC-7901 cells. Methods HMGB1 mRNA coding sequence was ampli- fied by reverse transcription polymerase chain reaction （RT-PCR） and inserted into pDsRED2-N1 vector between Xho I and EcoR I sites. The plasmid was transfected into SGC-7901 cells with lipofectamine, and red fluorescence protein DsRED2 was observed by fluorescent microscopy. HMGB1 protein expression was identified by Western blot. The effect of HMGB1 overexpression on oxaliplatin-induced apoptosis in SGC-7901 cells was evaluated by flow cytometry（Annexin-V/PI）. Results The eukaryotic expression vector pDsRED2-HMGB1 was constructed successfully. The expression of DsRED2-HMGB1 was observed in the transfected cells by fluorescent microscope. When treated with oxaliplatin, the apoptosis rate was significantly lower in pDsRED2-HMGBl-transfeeted SGC-7901 cells than that in pDsRED2-Nl-transfected SGC-7901 cells. Conclusion HMGB1 overexpression could inhibit oxaliplatin-induced apoptosis in SGC-7901 cells.

关 键 词：胃肿瘤 细胞凋亡 质粒 重组融合蛋白质类 

分 类 号：R735.2[医药卫生—肿瘤] R341.31[医药卫生—临床医学]