[MicroRNA-30a inhibits proliferation of hepatocellular carcinoma cells via targeted regulation of forkhead-box protein A1].

• DOI: 10.3760/cma.j.issn.1007-3418.2017.09.011
• Publication Date: 2017-09-20
• Authors: Yan Li; Lei Fu; Xiaoyu Fu; Ronghua Li; Shifang Peng
• Keywords: FOXA1; MTT assay

Objective: To investigate the expression of microRNA-30a (miR-30a) in hepatocellular carcinoma (HCC) and related molecular mechanisms regulating HCC cell proliferation. Methods: A total 30 pairs adjacent tissue samples were collected, quantitative real-time PCR Western blot used to measure mRNA protein forkhead-box A1 (FOXA1). Methyl thiazolyl tetrazolium (MTT) assay was evaluate proliferation cells, luciferase reporter gene performed verify target relationship between miR-30a FOXA1, MTT HepG2 cells FOXA1 after transfection. The t-test for comparison data two groups, a one-way analysis variance multiple groups. P < 0.05 considered statistically significant. Results: had significantly lower relative than (1.049 ± 0.380 vs 1.982 1.013, t = 3.985, 0.001). At 72 hours overexpression, there significant difference proliferative capacity blank control group, miR-30a-NC group (0.821 0.006 0.816 0.013 0.546 0.020, F 3.396, 0.05), suggesting that overexpression inhibited cells. its negatively regulated by miR-30a, activity wild-type mutant FOXA1-3'UTR vectors (1.221 0.024 2.658 0.031, 6.737, 0.05). In (1.019 0.016 1.022 0.017 0.227 0.021, 45.43, Upregulating reversed inhibitory effect on miR-30a+FOXA1 (0.524 0.023 0.843 0.019, 2.507, Conclusion: MiR-30a exerts FOXA1.目的： 探索微小RNA-30a（miR-30a）在肝细胞癌（HCC）中的表达及调控HCC细胞增殖的相关分子机制。 方法： 收集配对的HCC及癌旁组织30对，分别采用实时荧光定量PCR和Western印迹法检测叉头框蛋白A1（FOXA1）RNA和蛋白表达，四甲基偶氮唑蓝比色法（MTT）检测HepG2细胞增殖，萤光素酶报告基因检测验证miR-30a与FOXA1的靶点关系，MTT和Western blot法检测转染miR-30a后HepG2细胞增殖及FOXA1蛋白的表达。两组数据分析比较采用t检验，多组数据分析比较采用单因素方差分析，P 0.05为差异有统计学意义。 结果： miR-30a在HCC组织中的相对表达量为1.049±0.380显著低于癌旁组织的1.982±1.013，t 3.985，P 0.001,差异有统计学意义。过表达miR-30a h后，空白对照组HepG2细胞增殖能力为0.821±0.006，miR-30a-NC组为0.816±0.013，miR-30a组为0.546±0.020，过表达miR-30a能显著降低HepG2细胞的增殖，F 3.396，P 0.05，差异有统计学意义。FOXA1是miR-30a的靶基因，其蛋白表达被miR-30a负调控，荧光素酶的活性在野生型FOXA1-3'UTR载体中的表达量为1.221±0.024，在突变型FOXA1-3'UTR载体中的表达量为2.658±0.031，F 6.737，P 0.05，差异有统计学意义。过表达miR-30a能显著抑制FOXA1在HepG2细胞中的蛋白水平，FOXA1在空白对照组相对表达量为1.019±0.016，miR-30a-NC组为1.022±0.017，miR-30a组为0.227±0.021，F 45.43，P 0.05，差异有统计学意义。上调FOXA1的蛋白水平能够逆转miR-30a对HepG2细胞增殖的抑制作用，HepG2细胞的增殖能力在miR-30a组为0.524±0.023，在miR-30a+FOXA1组为0.843±0.019，t 2.507，P 0.05，差异有统计学意义。 结论： miR-30a对HCC细胞增殖的抑制作用是通过负调控FOXA1的表达而实现。.