摘要
目的探讨长链非编码RNA H19(long non-coding RNA H19,lncRNA H19)对miR-124-3p表达的影响及在肝癌细胞增殖中的作用。方法 Real-time PCR检测肝癌细胞系及临床标本中lncRNA H19和miR-124-3p的表达;在肝癌细胞系Hep G2和SMMC7721中过表达lncRNA H19,Real-time PCR检测miR-124-3p及其靶基因cyclin D1的表达;Western blot检测cyclin D1蛋白表达水平;CCK-8法检测lncRNA H19对细胞增殖的影响;过表达lncRNA H19同时加入antagomiR-124-3p后,检测miR-124-3p及其靶基因表达情况及细胞增殖情况。结果 lncRNA H19和miR-124-3p在肝癌细胞系及肝癌组织中均低表达(P<0.05);过表达lncRNA H19后miR-124-3p表达显著增加(P<0.05),miR-124-3p的靶基因cyclin D1蛋白和mRNA表达明显下降(P<0.05),细胞增殖受到抑制(P<0.05);过表达lncRNA H19同时干扰miR-124-3p后,细胞增殖抑制作用减弱(P<0.05)。结论长链非编码RNA H19通过促进miR-124-3p表达,进而抑制其靶基因cyclin D1表达而抑制肝癌细胞增殖。
Objective To determine the effect of long non-coding RNA H19 （lncRNA H19） on the expression of miR-124-3p and the proliferation of hepatocellular carcinoma cells. Methods Real-time PCR was used to detect the expression of lncRNA H19 and miR-124-3p in hepatocellular carcinoma cell lines （HepG2 and SMMC7721） and clinical specimens. After over-expressing lncRNA H19 in HepG2 and SMMC7721 cells, real-time PCR was carried out to detect the expression of miR-124-3p and its target gene, cyclinD1, and Western blotting was used to detect the expression of cyclinD1 at protein level. Cell proliferation was assessed by CCK-8 assay. After over-expressing lncRNA H19 and transferring antagomiR-124-3p into SMMC7721 and HepG2 cells in the meantime, the expression of cyclinD1 and the proliferation of cells were detected again. Results lncRNA H19 and miR-124-3p had low expression in tumor tissues and hepatocellular carcinoma cell lines （P〈0.05）. Compared with the control group, the expression of miR-124-3p was increased in SMMC7721 and HepG2 cells after over-expressing lncRNA H19 （P〈0.05）. The protein and mRNA levels of cyclinD1 were decreased in the over-expressing cells （P〈0.05）. Over-expression of lncRNA H19 suppressed the proliferation of HepG2 and SMMC7721 cells （P〈0.05）. However, overexpressing lncRNA H19 in the cells with antagomiR-124-3p transferring relieved the suppression of proliferation （P〈0.05）. Conclusion lncRNA H19 suppresses cyclinD1 expression and proliferation hepatocellular carcinoma cells through enhancing miR-124-3p.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2016年第6期554-559,共6页
Journal of Third Military Medical University
基金
国家自然科学基金面上项目(81571601)
国家自然科学基金地区科学基金项目(81360359)~~
