miR-15a-5p抑制人肝癌SMMC-7721细胞增殖与迁移

miR-15a-5p inhibits proliferation and migration of human hepatocellular carcinoma SMMC-7721 cells

目的:观察高表达的miR-15a-5p对人肝细胞癌SMMC-7721细胞增殖和迁移能力的影响。方法:化学合成加入EcoRⅠ和HindⅢ酶切位点的miR-15a-5p寡聚核苷酸,并进行测序确认;利用pcDNA6.2-GW/EmGFP质粒构建miR-15a-5p真核表达载体,瞬时转染SMMC-7721细胞,实时荧光定量PCR检测miR-15a-5p的表达;CCK-8法和台盼蓝染色活细胞计数检测SMMC-7721细胞的增殖能力,划痕实验检测细胞迁移能力的变化。结果:设计的miR-15a-5p序列与寡核苷酸测序结果匹配达100%;真核表达质粒瞬转后人肝癌SMMC-7721细胞miR-15a-5p的表达量与对照组相比显著增加(P<0.05);miR-15a-5p高表达SMMC-7721细胞的增殖能力与对照组相比均显著下降(P<0.05);miR-15a-5p高表达组细胞迁移速度低于对照组。结论:高表达的miR-15a-5p可抑制人肝癌SMMC-7721细胞的增殖和迁移能力。

AIM： To observe the influence of high expression of miR-15a-5p on the proliferation and migration of human hepatocellular carcinoma SMMC-7721 cells. METHODS： The miR-15a-5p oligonucleotide, which was recon-structed with additional restriction sites of EcoK I and Hind IH , was chemically synthesized and confirmed by sequencing.The miR-15a-5p eukaryotic expression system was constructed by pcDNA6. 2-GW/Em-GFP-pre-miR-15a-5p plasmid. The miR-15a-5p was transfected into the SMMC-7721 cells transiently by plasmid, and quantified by quantitative real-time PCR at the mRNA level. The cell viability was measured by CCK-8 assay, and the living cell counting was performed by the method of Trypan blue exclusion. The migration ability of the SMMC-7721 cells with high expression of miR-15a-5p was de-tected by wound healing test. RESULTS： The sequence of miR-15a-5p oligonucleotide 100% matched the designed se-quence. Compared with control group, the miR-15a-5p expression was increased significantly （P 〈0. 05）. The viability, the living cell number and the migration ability of the SMMC-7721 cells were decreased in high expression of miR-15a-5p group with statistically significant difference （P 〈 0. 05 ） . CONCLUSION ： The abilities of proliferation and migration in human hepatocellular carcinoma SMMC-7721 cells are decreased by high expression of miR-15a-5p.