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 ofhepatoma cells by targeting estrogen receptor α

作　　者：颜艳[1] 王琴[1] 李巍[1] 李旭[1] 阚鹏程[1] 刘梦圆[2] 

机构地区：[1]天津市环湖医院检验科,300060 [2]天津市神经外科研究所

出　　处：《中华肝脏病杂志》2015年第7期527-532,共6页Chinese Journal of Hepatology

基　　金：国家自然科学基金青年基金资助项目（81301967）

摘　　要：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-timeRT-PCR和Westernblot技术验证miR-211对其靶基因的直接调控作用I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p=6．26，P〈0．01）；miR-211增强肝癌细胞的侵袭能力忙值分别为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（f值分别为8．99、29．31，（D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。Objective To investigate the regulatory functions and molecular mechanisms of miR- 211 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. QGY- 7703 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 ct （ESR1） was identified and validated as a target gene of miR-211; knockdown of ESR1 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 ESR1.

关 键 词：微RNAS 肝肿瘤 雌激素受体Α 

分 类 号：R735.7[医药卫生—肿瘤]