【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。更多还原

【Abstract】 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.更多还原