上调miR-125b对肝癌Huh-7细胞系增殖及顺铂诱导凋亡的作用研究  

Upregulated miR- 125b changed Huh- 7 proliferation and apoptosis to cisplatin

作　　者：罗海峰[1] 杜渐[1] 张隽开[1] 巩鹏[1] 谭广[1] 王洪江[1] 

机构地区：[1]大连医科大学附属第一医院普外二科,辽宁大连116011

出　　处：《大连医科大学学报》2016年第1期12-15,共4页Journal of Dalian Medical University

基　　金：辽宁省自然科学基金项目(2013023045)

摘　　要：目的研究mi R-125b在肝癌发生发展中的作用机制,及其表达对于肝细胞癌化疗耐受性的影响。方法采用q PCR检测肝癌患者癌组织、癌旁组织、Huh-7细胞系(对照组)及转染mi R-125b mimic的Huh-7细胞系(转染组)中mi R-125b表达情况。采用流式细胞仪及MTT法检测对照组及转染组细胞周期及增殖情况。Western blot检测Huh-7细胞系转染后Mcl-1蛋白表达。设3组:对照组为正常培养细胞,miRNA control组转染miRNA control,mi R-125b mimic组转染mi R-125b mimic。Western blot检测Mcl-1特异性si RNA阻断Mcl-1蛋白表达后,细胞凋亡相关蛋白caspase3的表达情况。设3组:对照组细胞正常培养;顺铂组加入顺铂,终浓度50μg/m L培养;转染+顺铂组转染Mcl-1-si RNA,48 h后加入顺铂,终浓度50μg/m L。结果肝癌组织mi R-125b表达较癌旁组织明显减低,为癌旁组织的(46.24±8.53)%,P<0.05。mi R-125b mimic转染Huh-7细胞后,转染组细胞G1/S比例较对照组明显增高(P<0.05);Mcl-1蛋白表达较对照及miRNA control组下调(P<0.05);转染+顺铂组中的Caspase3蛋白表达较对照组及顺铂组明显增高(P<0.05)。结论上调mi R-125b的表达可以抑制肝癌细胞增殖,抑制靶蛋白Mcl-1表达,促进顺铂诱导凋亡,具有抑癌基因的作用。Objective Investigate mi R- 125 b function in hepatocellular carcinoma（ HCC） and its relation to chemoresistance. Methods 10 cases HCC specimen from clinic were chosen for detecting the mi R- 125 b expression by q RT-PCR,same detections were performed in 3 HCC cell lines as well. Transfected Huh- 7 cell line with mi R- 125 b mimic,detected cell proliferation by TUNEL,Mcl- 1 protein expression by western blot,and detected Caspase 3 expression when Mcl- 1- si RNA and cisplatin were added in Huh- 7 cells. Results HCC specimen mi R- 125 b expression was（ 46. 24 ± 8. 53） % of the specimen adjacent to cancer（ P 〈0. 05）. When mi R- 125 b was up- regulated,the ratio of G1/ S increased significantly compare to normal mi R- 125 b in Huh- 7 cells,Mcl- 1 protein expression were decreased compare to control（ P 〈0. 05）,Caspase 3 expression in Huh- 7 increased significantly after Mcl- 1- si RNA and cisplatin were added（ P 〈0. 05）. Conclusion mi R- 125 b suppressed the proliferation of Huh- 7,decreased Mcl- 1 protein expression and magnified HCC apoptosis to cisplatin. mi R- 125 b did as a cancer suppressor gene in HCC and possibly correlated with HCC chemoresistance.

关 键 词：MIRNA 肝细胞癌 化疗耐药性 

分 类 号：R735.7[医药卫生—肿瘤]