微小RNA-211靶定雌激素受体α促进肝癌细胞侵袭的研究
MiR-211 promotes invasion of hepatoma cells by targeting estrogen receptor α
目的 研究微小RNA (miR)-211对肝癌细胞恶性表型的影响,鉴定其下游直接靶基因,探讨miR-211调控肝癌细胞的分子机制. 方法 用实时荧光定量RT-PCR技术检测20对肝癌及癌旁组织中miR-211的表达水平;用Transwell侵袭实验检测miR-211对肝癌细胞QGY-7703及HepG2侵袭能力的影响;通过生物信息学方法,筛选miR-211可能的靶基因,并利用荧光报告载体实验结合Real-time RT-PCR和Westem blot技术验证miR-211对其靶基因的直接调控作用;用Transwell侵袭实验研究靶基因的具体功能.两组间差异比较用t检验. 结果 miR-211在肝癌组织中表达明显上调(t=6.26,P<0.01);miR-211增强肝癌细胞的侵袭能力(t值分别为12.59、17.82,P值均<0.01).筛选并证实了miR-211发挥作用的直接靶基因,雌激素受体α,且敲降雌激素受体α可促进肝癌细胞的侵袭(t值分别为8.99、29.31,P值均<0.01).结论 miR-211可靶定雌激素受体α而促进肝癌细胞的侵袭能力.
更多Objective To investigate the regulatory functions and molecular mechanisms of miR211 in hepatocellular carcinoma (HCC).Methods Real-time reverse transcription-PCR was used to analyze the expression of miR-211 in 20 paired clinical specimens of HCC and adjacent noncancerous tissues.QGY7703 and HepG2 cells with stimulation or inhibition of miR-211 expression were used to evaluate the effects on malignant phenotypes with the transwell invasion assay.Candidate target genes of miR-211 were identified by bioinformatic screening and verified by the EGFP report assay,real-time PCR and western blotting.Moreover,the regulatory functions of miR-211 on the target genes were investigated by RNA interference and cell phenotype assays.Results miR-211 was up-regulated in HCC tissue specimens (t =6.26,P < 0.01).HCC cells overexpressing miR-211 showed greater invasive capacity than cells with inhibited expression (QGY-7703:t =12.59,P < 0.01;HepG2:t =17.82,P < 0.01).Estrogen receptor α (ESR1) was identified and validated as a target gene of miR-211;knockdown of ESR 1 promoted HCC invasive capacity (QGY-7703:t =8.97,P < 0.01;HepG2:t =29.31,P < 0.01).Conclusion miR-211 promotes invasion of carcinoma cells by directly targeting ESR 1.
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