miR-495在肝癌组织中的表达及其对肝癌MHCC-97H细胞的影响

Expression of miR-495 and its effect on MHCC-97H hepatocellular carcinoma cells

目的探讨miR-495在肝癌组织中的表达以及对肝癌MHCC-97H细胞的影响。方法选取2017年1月至2018年1月我院保存的肝癌组织标本56例(肝癌组),同时选取正常肝脏组织标本40例作为对照组,采用逆转录实时荧光定量PCR(qRT-PCR)检测miR-495表达;选取肝癌MHCC-97H细胞,随机分为对照组、空白质粒组和转染组,其中空白质粒组转染空白质粒,转染组转染miR-495抑制剂,用qRT-PCR检测各组细胞miR-495表达,用CCK法检测各组细胞增殖活性,用Transwell细胞迁移实验检测各组细胞迁移能力。多组间数据比较使用方差分析,进一步两两比较采用LDS-t检验,两组比较使用t检验。结果肝癌组miR-495相对表达量为2.043±0.382,高于对照组,两组比较,差异有统计学意义(P<0.05);临床分期Ⅲ~Ⅳ期、有淋巴结转移患者miR-495相对表达量分别为2.265±0.284和2.290±0.355,高于Ⅰ~Ⅱ期、无淋巴结转移患者(P<0.05),差异有统计学意义;转染组miR-495相对表达量为0.653±0.102,低于对照组和空白质粒组(P<0.05),差异有统计学意义;转染组培养24 h和48 h MHCC-97H细胞A值分别为0.404±0.106和0.604±0.136,低于对照组和空白质粒组(P<0.05);转染组MHCC-97H细胞迁移数为(6.10±1.20)个,低于对照组和空白质粒组(P<0.05)。结论miR-495在肝癌组织中呈高表达,与临床病理特征有一定关系;miR-495对肝癌MHCC-97H细胞增殖以及迁移能力有一定影响。

Objective To investigate the expression of miR-495 and its effect on MHCC-97H hepatocellular carcinoma cells.Methods Fifty-six hepatocellular carcinoma tissue specimens(HCC group)and 40 normal liver tissue specimens(control group)preserved in our hospital from January 2017 to January 2018 were selected.Reverse transcription real-time fluorescent quantitative PCR(qRT-PCR)was used for miR-495 expression detection.MHCC-97H HCC cells were randomly selected and then divide into control group,blank plasmid group and transfection group.The blank plasmid group was transfected with blank plasmid,and the transfection group was transfected with miR-495 inhibitor.The expression of miR-495 in each group of cells were detected using qRT-PCR.CCK method was used to detect each group proliferation activity.Transwell cell migration assay was used to detect each group migration ability.Analysis of variance was used for comparison between multiple groups.Furthermore,LDS-t test was used for pairwise comparison,and t-test was used for comparison between the two groups.Results The relative expression levels of miR-495 in the HCC group was(2.043±0.382),which was higher than the control group,and the difference between the two groups was statistically significant(P<0.05).The relative expressions levels of miR-495 in patients with stage III to IV and lymph node metastasis were 2.265±0.284 and 2.290±0.355,which were significantly higher than those of stage I to II and no lymph node metastasis(P<0.05).The relative expression levels of miR-495 in transfection group was 0.653±0.102,which were significantly lower than control group and blank plasmid group(P<0.05).The A values of MHCC-97H cells cultured for 24 h and 48 h in transfection group were 0.404±0.106 and 0.604±0.136,which were significantly lower than control group and blank plasmid group(P<0.05).MHCC-97H cells migration number in the transfection group was(6.100±20),which was significantly lower than that of control group and blank plasmid group(P<0.05).Conclusion miR-495 high expression has certain relationship with clinicopathological characteristics of HCC tissues.In addition,miR-495 has a certain effect on the proliferation and migration ability of MHCC-97H HCC cells.