化学乏氧人肝癌细胞SMMC-7721中HIF-1α和miR-210的表达  

Expression of Hypoxia Induced Factor-1α and miR-210 in Chemical Hypoxic Human Hepatoma SMMC-7721

作　　者：杨巍[1] 朱巍[1] 曹建平[1] 刘芬菊[1] 

机构地区：[1]苏州大学放射医学与公共卫生学院放射生物学教研室,江苏苏州215123

出　　处：《苏州大学学报（医学版）》2011年第5期694-697,共4页Suzhou University Journal of Medical Science

基　　金：国家自然科学基金青年项目(30600160);国家自然科学基金面上项目(81071958);苏州大学引进人员启动资助项目(Q4126803);教育部"长江学者和创新团队发展计划项目"(IRT0849)

摘　　要：目的探讨不同浓度CoCl2对人肝癌细胞SMMC-7721乏氧诱导因子-1α(HIF-1α)和miR-210表达的影响以及RNA干扰HIF-1α基因对miR-210表达的影响。方法采用Western blot法检测不同浓度CoCl2培养24 h后SMMC-7721细胞HIF-1α蛋白表达变化,实时定量PCR检测miR-210相对表达量。脂质体介导靶向HIF-1α基因的RNA干扰质粒pGenesil-HIF转染SMMC-7721细胞后,乏氧培养24、48和72 h,分别采用Western blot法和实时定量PCR检测细胞HIF-1α蛋白表达变化和miR-210相对表达量。结果 50和100μmol/L CoCl2培养24 h后细胞HIF-1α蛋白表达和miR-210相对表达量与0μmol/L组相比,差异均无统计学意义(均P>0.05);150和200μmol/L CoCl2培养24 h后细胞HIF-1α蛋白表达和miR-210相对表达量均显著高于0μmol/L组(P<0.05或P<0.01)。转染质粒pGenesil-HIF后乏氧培养24、48和72 h,细胞HIF-1α蛋白表达和miR-210相对表达量显著低于阴性干扰组(均P<0.01)。结论化学乏氧人肝癌细胞SMMC-7721中HIF-1α和miR-210表达明显上调,RNA干扰HIF-1α基因可明显下调乏氧细胞miR-210表达。Objective To investigate the effect of CoCl2 concentration on the expression of hypoxia induced factor-1α and miR-210 in chemical hypoxic human hepatoma SMMC-7721 and the effect of RNA interference of HIF-1α gene on miR-210 expression.Methods Expression of HIF-1α gene and miR-210 was detected by Western blot and real-time quantitative PCR in SMMC-7721 cells cultured in various concentration of CoCl2 for 24 h respectively.SMMC-7721 cells were cultured in hypoxic condition for 24,48 and 72 h after transfection with pGenesil-HIF mediated by liposome,expression of HIF-1α gene and miR-210 of which was detected by Western blot and real-time quantitative PCR respectively.Results Expression of HIF-1α gene and miR-210 showed no significant difference in SMMC-7721 cells cultured in 50 and 100 μmol/L CoCl2 for 24 h compared to that of 0 μmol/L CoCl2.Expression of HIF-1α gene and miR-210 was significantly higher in SMMC-7721 cells cultured in 150 and 200 μmol/L CoCl2 for 24 h than that in 0 μmol/l CoCl2（P0.05 or P0.01）.SMMC-7721 cells were cultured in hypoxic condition for 24,48 and 72 h after transfection with pGenesil-HIF,expression of HIF-1α gene and miR-210 of which was significantly lower than that in the negative interference group（P0.01）.Conclusion Expression of HIF-1α and miR-210 was up-regulated and RNA interference of HIF-1α gene could down-regulate miR-210 expression in chemical hypoxic human hepatoma SMMC-7721.

关 键 词：乏氧诱导因子-1Α MIR-210 肝癌 RNA干扰 

分 类 号：R730.5[医药卫生—肿瘤]