摘要
目的:探究微小RNA-92a-3p(miR-92a-3p)对肝细胞癌(HCC)的影响及其分子机制。方法:qRT-PCR检测人肝细胞癌细胞系SMMC-7721、Bel-7402、HepG2、Hep3B和人正常肝细胞系HL-7702中miR-92a-3p和Krüppel样因子4(KLF4)的表达。将SMMC-7721、Bel-7402细胞分组为Inhibitor NC组、miR-92a-3p Inhibitor组、mimic NC组、miR-92a-3p mimic组、mimic NC+oe-NC组、mimic NC+oe-KLF4组、miR-92a-3p mimic+oe-NC组、miR-92a-3p mimic+oe-KLF4组。qRT-PCR检测细胞中miR-92a-3p和KLF4的mRNA表达水平。Western blot检测KLF4蛋白表达水平。MTT增殖实验检测细胞活力。划痕愈合、Transwell侵袭实验分别检测细胞迁移能力和侵袭能力。通过TargetScan分析miR-92a-3p与KLF4的靶向关系,并通过双荧光素酶实验验证。结果:相比于正常细胞,miR-92a-3p在HCC细胞中高表达,KLF4在HCC细胞中低表达(P<0.05)。与正常表达组相比,下调miR-92a-3p显著抑制了SMMC-7721和Bel-7402细胞的增殖、迁移、侵袭能力(P<0.05)。双荧光素酶实验验证了miR-92a-3p与KLF4存在靶向结合位点。过表达KLF4能够阻碍HCC细胞的增殖、迁移和侵袭,抑制过表达miR-92a-3p对HCC细胞生长的促进作用,差异有统计学意义(P<0.05)。结论:miR-92a-3p能够通过靶向下调KLF4来促进HCC细胞的增殖、迁移、侵袭。
Objective:To investigate the effect of microRNA-92a-3p(miR-92a-3p)on hepatocellular carcinoma(HCC)and its molecular mechanism.Methods:qRT-PCR was used to detect the expression of miR-92a-3p and Krüppel-like factor 4(KLF4)in human HCC cell lines SMMC-7721,Bel-7402,HepG2,Hep3B,and human normal liver cell line HL-7702.SMMC-7721 and Bel-7402 cells were divided into eight groups:Inhibitor NC group,miR-92a-3p Inhibitor group,mimic NC group,miR-92a-3p mimic group,mimic NC+oe-NC group,mimic NC+oe-KLF4 group,miR-92a-3p mimic+oe-NC group,and miR-92a-3p mimic+oe-KLF4 group.qRT-PCR was used to detect the mRNA expression levels of miR-92a-3p and KLF4 in cells.Western blot was used to detect the protein expression levels of KLF4.MTT proliferation assay was used to detect cell viability.Scratch healing and Transwell invasion assays were used to detect cell migration and invasion abilities,respectively.TargetScan was used to analyze the targeting relationship between miR-92a-3p and KLF4,and the results were verified by dual luciferase assay.Results:Compared with normal cells,miR-92a-3p was highly expressed in HCC cells,and KLF4 was lowly expressed in HCC cells(P<0.05).Compared with the normal expression group,down-regulation of miR-92a-3p significantly inhibited the proliferation,migration,and invasion abilities of SMMC-7721 and Bel-7402 cells(P<0.05).The dual luciferase assay verified the existence of a targeting binding site between miR-92a-3p and KLF4.Overexpression of KLF4 was able to inhibit the proliferation,migration,and invasion of HCC cells,and inhibit the promoting effect of overexpressed miR-92a-3p on HCC cell growth,with statistical significance(P<0.05).Conclusion:miR-92a-3p can promote proliferation,migration,and invasion of HCC cells by targeting down-regulation of KLF4.
作者
蔡青山
郑建兴
申月玲
吴东洋
李树栋
刘立友
刘东
CAI Qingshan;ZHENG Jianxing(Tangshan Central Hospital,Hebei Tangshan 063000,China)
出处
《河北医学》
CAS
2024年第2期199-204,共6页
Hebei Medicine
基金
2022年度河北省医学科学研究课题计划,(编号:20221840)。
作者简介
通讯作者:郑建兴。
