微小RNA-34a靶向X连锁凋亡抑制蛋白基因对肝癌细胞增殖的影响  

Effect of microRNA-34a and its target gene X-linked inhibitor of apoptosis protein on proliferation activity and cell apoptosis of hepatocarcinoma cells

作　　者：张正威[1] 卢忠心[2] 刘水逸[2] 孔晓宇[3] 王静[2] 魏礼清[2] 

机构地区：[1]华中科技大学同济医学院附属武汉中心医院综合科,武汉430014 [2]华中科技大学同济医学院附属武汉中心医院检验科,武汉430014 [3]华中科技大学同济医学院附属武汉中心医院肝胆外科,武汉430014

出　　处：《中华实验外科杂志》2015年第7期1622-1624,共3页Chinese Journal of Experimental Surgery

基　　金：武汉市中心医院青年基金项目（YQ14A02）

摘　　要：目的 观察微小RNA-34a（miR-34a）及其靶基因X连锁凋亡抑制蛋白（XIAP）对肝癌细胞增殖活力和细胞凋亡的影响及其能机制.方法 通过脂质体将人工合成的miR-34a模拟物及其阴性对照以终质量浓度200 nmol/L转染人肝癌SMMC7721细胞,采用细胞计数试剂盒-8（CCK-8）检测转染后SMMC7721细胞的增殖活力,采用流式细胞术检测细胞凋亡,并进一步采用实时荧光定量聚合酶链反应（FQ-PCR）和Western blot分析XIAP的表达水平.结果 转染miR-34a模拟物的实验组细胞的增殖抑制率在24、48、72 h均显著高于对照组细胞（P＜0.01）,并且在72h转染组细胞凋亡率（6.98±0.13）％高于阴性对照组（4.89 ±0.12）％和空白组（4.02±0.21）％（P＜0.05）.miR-34a模拟物转染SMMC7721细胞后,XIAP蛋白在空白组、转染组和阴性对照组的相对表达量分别为0.942±0.028、0.489±0.033和0.982±0.032,转染组较其他两组明显降低（P＜0.05）.结论 miR-34a可抑制肝癌细胞增殖,其机制可能与其下调XIAP的表达有关.Objective To investigate the effect of microRNA-34a （miR-34a） on X-linked inhibitor of apoptosis protein （XIAP） and its role in the proliferation activity and cell apoptosis of hepatocarcinoma cells.Methods Synthetic miR-34a mimic and its negative control were transfected into human hepatocarcinoma SMMC7721 cells to a final concentration of 200 nmol/L by liposome method.After transfection,the cell counting kit-8 （CCK-8） was used to measure the cell proliferation activity.The cell apoptosis was determined by flow cytometry.The expression of XIAP mRNA and protein was detected by real-time fluorescent quantitative polymerase chain reaction （FQ-PCR） and Western blotting respectively.Results The transfection efficiency mediated by cationic liposome was greater than 85％.The inhibitory rate of cell proliferation in SMMC7721 cells was （39.5 ± 10.5）％,（43.5 ± 2.8）％ and （49.6 ± 0.3） ％ at 24,48 and 72 h respectively after transfection with miR-34a mimic,which was significantly increased as compared with the control cells.Moreover,the cell apoptosis in experimental group （6.98 ± 0.13）％ was higher than that in negative control group （4.89 ± 0.62）％ and simple liposome group （4.02±0.21）％ at 72 h.Whereas the expression of XIAP protein was decreased in SMMC7721 cells transfected with miR-34a mimic at 72 h after transfection.Conclusion miR-34a suppresses the proliferation activity of hepatocarcinoma cells by down-regulating XIAP gene expression.

关 键 词：癌 肝细胞 微小RNA-34a X连锁凋亡抑制蛋白 

分 类 号：R541.4[医药卫生—心血管疾病]