摘要
目的探讨长链非编码RNA-H19(lncRNA-H19)能否调控人肝癌细胞HepG2增殖和凋亡。方法以人肝癌细胞株HepG2作为转染对象,分别设空白对照组(人肝癌细胞HepG2+培养基)、阴性对照组(人肝癌细胞HepG2+培养基+空载体)和沉默组(人肝癌细胞HepG2+培养基+慢病毒);采用含10%胎牛血清的RPMI-1640培养基,培养人肝癌细胞HepG2,使用慢病毒转染技术沉默人肝癌细胞HepG2的lncRNA-H19表达,荧光实时定量聚合酶链反应(qRT-PCR)检测人肝癌细胞HepG2的lncRNA-H19表达;四甲基偶氮唑蓝(methyl thiazolyl tetrazolium,MTT)法检测人肝癌细胞HepG2增殖能力;流式细胞术检测人肝癌细胞HepG2凋亡率。结果 qRT-PCR检测lncRNA-H19的相对表达量,三组比较差异具有统计学意义(F=383.961,P<0.01);沉默lncRNA-H19后,用MTT法分别于24h、48h和72h检测人肝癌细胞HepG2吸光度的表达量,三组比较差异具有统计学意义(F=266.198,P<0.05),其抑制作用72h>48h>24h,表现为时间依赖;沉默lncRNA-H19后,3次流式细胞术检测细胞凋亡率均数,三组比较差异具有统计学意义(F=68.823,P<0.001)。表明沉默lncRNA-H19可促进肝癌细胞HepG2细胞增殖,抑制细胞凋亡。结论Ln-cRNA-H19可以调控人肝癌HepG2细胞的生长,能抑制肝癌细胞HepG2的增殖,其抑制作用呈时间依赖,促进HepG2细胞凋亡。
Objective To investigate whether long non-coding RNA H19(lncRNA-H19)can regulate the proliferation and apoptosis of human hepatoma HepG2 cell lines.Methods Human hepatoma cell line was used as transfection target,and the cells were divided into a blank control group(human hepatoma cell HepG2+medium),a negative control group(human hepatoma cell HepG2+medium+empty vector)and a silent group(human hepatoma cell HepG2+medium+lentivirus).Human hepatoma HepG2 cells were cultured in RPMI-1640 medium containing 10%fetal bovine serum,and lncRNA-H19 expression of human hepatoma cell line HepG2 was silenced by lentiviral transfection technique,and real-time quantitative polymerase chain reaction(qRT-PCR)was used to detect the expression of lncRNA-H19 in human hepatoma HepG2 cells.Methyl thiazolyl tetrazolium(MTT)was used to detect the proliferation in human hepatoma HepG2 cells;human hepatoma HepG2 cell apoptosis was detected by flow cytometry.Results The relative expression of lncRNA-H19 detected by qRT-PCR differed statistically by comparing among the three groups(F=383.961,P<0.01).After silencing the lncRNA-H19 gene,the expression of human hepatoma HepG2 cell absorbance was measured by MTT in 24 hours,48 hours and 72 hours,respectively,and there was statistical difference by comparing among the three groups(F=266~198,P<0.05).The inhibitory effect showed a time-dependent manner and ranged as the following:72 hours >48 hours >24 hours.After silencing lncRNA-H19,the average rate of apoptosis was detected by flow cytometry for 3 times and there was statistical difference among the three groups(F=68.823,P<0.001).Results showed that silencing lncRNA-H19 could promote the proliferation of hepatoma HepG2 cells and inhibit cell apoptosis.Conclusion LncRNA-H19 can regulate the growth of human hepatoma HepG2 cells and inhibit the proliferation of HepG2 cells in a time-dependent manner,which promotes the apoptosis of HepG2 cells.
作者
黎梨
邓凤莲
黄赞松
廖俊
李建基
杨哲
Li Li;Deng Fenglian;Huang Zansong;Liao Jun;Li Jianji;Yang Zhe(Affiliated Hospital of Youjiang Medical University for Nationalities,Guangxi Clinical Medicine Centre for Liver Disease,Baise 533000,Guangxi,China;Graduate School of Youjiang Medical University for Nationalities,Baise 533000,Guangxi,China)
出处
《右江民族医学院学报》
2019年第1期15-19,25,共6页
Journal of Youjiang Medical University for Nationalities
基金
广西自然科学基金资助项目(桂财教2014GXNSFAA118143)
2017年度广西医药卫生自筹经费计划课题(桂卫Z20170224)
广西科技基地与人才专项(广西肝胆疾病临床医学研究中心研究课题)(桂科AD17129035)
2017年广西研究生教育创新计划项目(桂学位YCSW2017214)
广西高校桂西地区高发病防治重点实验室开放课题(kfkt2017006)
关键词
lncRNA
人肝癌HEPG2细胞
肝肿瘤
实验性
增殖
凋亡
lncRNA
human hepatocellular carcinoma HepG2 cells
liver neoplasms,experimental
roliferation
apoptosis
