miR-29a靶向调控CLDN1抑制肝癌细胞增殖的实验研究

目的:探讨miR-29a在肝癌组织中的表达水平，及其对人肝癌细胞增殖能力的影响。方法:通过RT-PCR测定肝癌组织及非肝癌组织中miR-29a的表达水平；将携带miR-29a的脂质体miR-29a mimic、空载体miR-NC以及miR-29a抑制剂anti-miR-29a转染入肝癌细胞系HepG2和HLE，通过RT-PCR方法检测miR-29a对肝癌细胞系HepG2、HLE增殖能力影响；通过Targetscan软件预测miR-29a靶基因，采用Western blot及荧光素酶实验验证。结果:与非肝癌组织比较，miR-29a在肝癌组织中表达下调，MTT实验提示:miR-29a高表达组肝癌细胞增殖能力较弱，miR-29a低表达组肝癌细胞增殖能力提高。Targetscan软件提示:CLDN1可能为miR-29a靶基因，Western blot及荧光素酶实验提示:与阴性对照组相比，miR-29a高表达组内CLDN1表达下降，相反miR-29a低表达组内CLDN1表达增加。结论:miR-29a在肝癌细胞及组织中表达下调，并且可能通过调控CLDN1影响肝癌细胞增殖能力。

Experimental study on the inhibitory effect of miR-29a regulatory CLDN1 on the proliferation of hepatocellular carcinoma cells

Objective:To explore the expression level of miR-29a in liver cancer tissue and its effect on the proliferation ability of human liver cancer cells.Methods:The expression of miR-29a in hepatocellular carcinoma and non-hepatocellular carcinoma tissues were detected by RT-PCR.miR-29a mimics,empty vector miR-29a-NC and miR-29a inhibitors were transfected into hepatocellular carcinoma HepG2 and HLE cell lines, RT-PCR methods were used to observe the effects of miR-29a on proliferation of hepatocellular carcinoma cell lines HepG2 and HLE. The target genes of miR-29a were predicted by Targetscan software and verified by Western blot and luciferase experiments.Results:Compared with non-hepatocarcinoma tissues,miR-29a was down-regulated in hepatocarcinoma tissues.MTT experiments showed that the proliferation ability of hepatoma cells in the high expression group of miR-29a were weak.The proliferation ability of hepatoma cells in the low expression group of miR-29a were increased.Targetscan software suggested that CLDN1 may be a target gene of miR-29a.Western blot and luciferase assays showed that the expression of CLDN1 was decreased in the high expression group of miR-29a compared with the negative control group,and the expression of CLDN1 was decreased in the low expression group of miR-29a.Conclusion:miR-29a is down-regulated in liver cancer cells and tissues,and may affect the proliferation of liver cancer cells by regulating CLDN1.