摘要目的:观察微小RNA(miR)-142-3p靶向CD133对肝癌细胞增殖、侵袭和干细胞特性的影响。方法:选取郑州大学第二附属医院2017年6月至2019年12月手术切除的肝癌组织和对应的癌旁组织作为研究对象。采用荧光定量聚合酶链反应(PCR)分析癌旁组织和肿瘤组织miR-142-3p表达水平。采用慢病毒在MHCC97H肝癌细胞建立miR-142-3p过表达和对照细胞系(miR-142-3p组和对照组)，采用细胞计数试剂盒(CCK-8)和EdU染色分析两组细胞增殖；采用Transwell分析两组细胞侵袭；采用软琼脂克隆形成实验分析两组细胞干细胞特性；生物信息学和双荧光素酶报告基因分析miR-142-3p靶基因；采用蛋白质印迹法(Western blot)分析两组增殖、侵袭和干细胞标志物细胞核增殖抗原(Ki-67)、基质金属蛋白酶(MMP)-9、CD133和Nanog蛋白表达水平，组间比较采用 t检验。 结果:miR-142-3p组细胞miR-142-3p表达水平(0.41±0.12)显著低于癌旁组织miR-142-3p表达水平(1.06±0.25)，差异有统计学意义( t＝3.081， P<0.05)。miR-142-3p组细胞吸光度( A)值(1.63±0.20)低于对照组细胞 A值(2.13±0.22)，差异有统计学意义( t＝2.918， P<0.05)。miR-142-3p组细胞EdU染色阳性率[(30.48±5.12)%]低于对照组细胞EdU染色阳性率[(87.89±7.28)%]，差异有统计学意义( t＝3.409， P<0.05)。miR-142-3p组细胞Ki-67蛋白表达水平(0.50±0.13)低于对照组细胞Ki-67表达水平(1.15±0.14)，差异有统计学意义( t＝2.319， P<0.05)。miR-142-3p组细胞侵袭数量[(67.59±6.01)个]低于对照组细胞侵袭数量[(121.35±6.12)个]，差异有统计学意义( t＝3.409， P<0.05)。miR-142-3p组细胞MMP-9蛋白表达水平(0.48±0.18)低于对照组细胞MMP-9蛋白表达水平(1.57±0.15)，差异有统计学意义( t＝2.917， P<0.05)。miR-142-3p组细胞克隆形成率[(31.44±4.51)%]低于对照组细胞克隆形成率[(57.68±7.19)%]，差异有统计学意义( t＝5.103， P<0.05)。miR-142-3p组细胞Nanog蛋白表达水平(0.51±0.14)低于对照组细胞Nanog蛋白表达水平(1.36±0.17)，差异有统计学意义( t＝2.514， P<0.05)。生物信息学和双荧光素酶报告基因显示CD133是miR-142-3p的靶基因。miR-142-3p组细胞CD133蛋白表达水平(0.43±0.11)低于对照组细胞CD133蛋白表达水平(1.15±0.13)，差异有统计学意义( t＝2.514， P<0.05)。 结论:miR-142-3p通过靶向调控CD133表达水平调节着肝癌细胞增殖、迁移和干细胞特性。

abstractsObjective:To investigate the effects of microRNA (miRNA, miR)-142-3p on the proliferation, invasion and stem cell characteristics of hepatoma cells by targeting CD133.Methods:Liver cancer tissues and corresponding adjacent tissues from June 2017 to December 2019 were selected as the research objects. The expression of miR-142-3p was analyzed by fluorescence quantitative polymerase chain reaction (PCR). miR-142-3p overexpression and control cell lines (miR-142-3p group and control group) were established by lentivirus in MHCC97H hepatoma cells. Cell proliferation was analyzed by cell counting kit-8 (CCK-8) and EdU staining. Cell invasion was analyzed by Transwell. Clone formation experiment was analyzed by soft agar; miR-142-3p target gene was analyzed by bioinformatics and double luciferase reporter gene. Proliferating cell nuclear antigen (Ki-67), matrix metalloproteinase (MMP)-9, CD133 and Nanog proteins were analyzed by Western blotting.Results:Compared with the expression level of miR-142-3p in adjacent tissues (1.06±0.25), the expression of miR-142-3p in tumor tissues was significantly decreased (0.41±0.12, t=3.081, P<0.05). Compared with the control group (2.13±0.22), the cell absorbance ( A) (1.63±0.20) in miR-142-3p group was significantly decreased ( t=2.918, P<0.05). Compared with the control group (87.89±7.28)%, the positive rate of EdU staining in miR-142-3p group (30.48±5.12)% was significantly reduced ( t=3.409, P<0.05). Compared with the control group (1.15±0.14), the expression of Ki-67 protein in miR-142-3p group was significantly decreased (0.50±0.13, t=2.319, P<0.05). Compared with the control group (121.35±6.12), the invasion number of miR-142-3p group (67.59±6.01) was significantly decreased ( t=3.409, P<0.05). Compared with the control group (1.57±0.15), the expression of MMP-9 protein in miR-142-3p group (0.48±0.18) was significantly decreased ( t=2.917, P<0.05). Compared with the control group [(57.68±7.19)%], the colony formation rate in miR-142-3p group [(31.44±4.51)%] was significantly decreased ( t=5.103, P<0.05). Compared with the control group (1.36±0.17), the expression of Nanog protein in miR-142-3p group (0.51±0.14) was significantly decreased ( t=2.514, P<0.05). Bioinformatics and double luciferase reporter gene showed that CD133 was the target gene of miR-142-3p. Compared with the control group (1.15±0.13), the expression of CD133 protein in miR-142-3p group (0.43±0.11) was significantly decreased ( t=2.514, P<0.05). Conclusion:MiR-142-3p regulates the proliferation, migration and stem cell characteristics of hepatoma cells by targeting the expression level of CD133.