MiR-194过表达和抑制表达对肝癌细胞株Hep-3b中侧群细胞增殖的影响  

Influence of miR-194 up and down-expression on liver cancer stem cells

作　　者：许扬梅[1] 龚福生[1] 刘沁颖[1] 刘施佳[1] 黄丽洁[1] 郑秋红[1] XU Yangmei;GONG Fusheng;LIU Qinying;LIU Shijia;HUANG Lijie;ZHENG Qiuhong(Fujian Provincial Key Laboratory of Tumor Biotherapy,Fujian Medical University CancerHospital&Fujian Cancer Hospital,Fuzhou,Fujian 350014,China)

机构地区：[1]福建医科大学附属肿瘤医院福建省肿瘤医院福建省肿瘤生物治疗重点实验室

出　　处：《福建医药杂志》2019年第6期141-145,F0004,共6页Fujian Medical Journal

基　　金：福建省自然科学基金资助项目(2016J01436)

摘　　要：目的研究MicroRNA-194(miR-194)过表达和抑制表达对细胞株Hep-3b中侧群细胞增殖功能影响。方法构建miR-194过表达和抑制表达慢病毒并感染Hep-3b细胞。流式细胞仪鉴定各感染组SP侧群比例,QPCR检测各组细胞miR-194-5p表达,CCK-8法检测各组细胞增殖能力,软琼脂克隆形成实验检测细胞克隆形成能力。结果流式细胞仪鉴定发现miR-194过表达Hep-3b组SP细胞含量与miR-194抑制表达Hep-3b组差异具有统计学意义(LSD-t=3.550,P=0.012)。QPCR检测显示miR-194过表达Hep-3b组miR-194-5p水平高于miR-194抑制表达Hep-3b组(LSD-t=4.680,P=0.007)。CCK-8实验表明miR-194过表达Hep-3b组和miR-194抑制表达Hep-3b组细胞增殖能力差异有统计学意义(LSD-t=5.621,P=0.002)。软琼脂克隆形成实验显示miR-194过表达Hep-3b组比miR-194抑制表达Hep-3b组形成细胞克隆少(LSD-t=5.849,P=0.001)。结论成功构建了miR-194过表达和抑制细胞株,miR-194抑制组显示明显的干细胞特征,其含有更多SP细胞,具有更强的细胞增殖能力和自我更新能力。Objective To study the influence of miR-194 up and down-expression Hep-3 b cell lines on the proliferation ability of SP cells.Methods Hep-3 b was used to construct miR-194 up and down-expression cell lines via lentivirus infection.Flow cytometry was applied to identify SP cells.QPCR assay was adopted to test the expression of miR-194-5 p in those cell lines.CCK-8 assay was conducted to study the proliferation ability of cells and soft agar colony formation assay was conducted to test colony-forming ability.Results Each infected cell line maintained good growth.Flow cytometry analysis showed that the SP percentage were significantly different between miR-194 up and down-regulation Hep-3 b groups(LSD-t=3.550,P=0.012).QPCR assay of miR-194-5 p showed statistic difference between miR-194 up and down-regulation Hep-3 b groups(LSD-t=4.680,P=0.007).CCK-8 assay showed different growth ability between miR-194 up and down-regulation Hep-3 b groups(LSD-t=5.621,P=0.002).Soft agar colony formation assay results less colonies formed in miR-194 up-regulation Hep-3 b group than miR-194 down-regulation Hep-3 b group(LSD-t=5.849,P=0.001).Conclusion miR-194 up-regulation Hep-3 b group than miR-194 down-regulation Hep-3 b group were successfully constructed.miR-194 down-regulation Hep-3 b group contains more SP cells,possess stronger growth ability and self-renewal ability,showing stronger proliferation and self-renewal ability.

关 键 词：微小RNA 慢病毒载体 肝癌 干细胞 

分 类 号：R735.7[医药卫生—肿瘤]