【摘要】 目的探讨miR-195过表达对口腔鳞状细胞癌(OSCC)细胞增殖的影响及作用机制。方法将培养好的SCC-4细胞随机分组,设计合成miR-195 mimic,并将对照试剂、miR-195 mimic试剂分别转染至对照组、miR-195 mimic组。用细胞活力测定(MTS)实验检测miR-195过表达对细胞增殖的影响;用生物信息软件及荧光素酶报告基因实验检测miR-195对成纤维细胞生长因子7(FGF7)基因的靶向调控作用;qRT-PCR检测miR-195过表达对细胞内FGF7 mRNA表达的影响。构建FGF7过表达质粒,将FGF7过表达对照vector+miR-195 mimic试剂、FGF7过表达+miR-195 mimic试剂分别转染至miR-195 mimic+vector组、miR-195 mimic+FGF7组,MTS实验检测FGF7对miR-195 mimic SCC-4细胞增殖的影响,Western blotting实验检测细胞内磷酸化磷脂酰肌醇3-激酶(pPI3K)、磷酸化蛋白激酶B(pAKT)蛋白的表达。结果细胞贴壁第3、4天miR-195 mimic组细胞增殖能力较对照组低(P均<0.05)。生物信息软件预测和荧光素酶报告基因实验证实,FGF7为miR-195的靶基因。细胞贴壁第2、3、4天miR-195 mimic+FGF7组细胞增殖能力较miR-195 mimic+vector组高(P均<0.05)。miR-195 mimic组pI3K、pAKT蛋白相对表达量低于对照组(P均<0.05),miR-195 mimic+FGF7组pI3K、pAKT蛋白相对表达量高于miR-195 mimic组(P均<0.05)。结论 miR-195过表达可抑制DSCC细胞增殖,其作用机制可能为通过靶向下调FGF7表达以降低PI3K/AKT信号通路活性。更多还原

【Abstract】 Objective To investigate the effect and mechanism of microRNA-195(miR-195) on the proliferation of oral squamous cell carcinoma(OSCC) cells. Methods The cultured OSCC cells were randomly divided into two groups: the control group and miR-195 mimic group. The miR-195 mimic was designed and synthesized. The control reagent and miR-195 mimic were transfected into the control group and miR-195 mimic group. The effect of miR-195 on cell proliferation was detected by MTS assay. The bio-information software and luciferase reporter gene assay were used to detect the targeted regulation of miR-195 on fibroblast growth factor 7(FGF7) gene; qRT-PCR was used to detect the effect of miR-195 on the expression of FGF7 mRNA in cells; FGF7 overexpression plasmid was constructed, and FGF7 overexpression control vector+miR-195 mimic reagent and FGF7 overexpression+miR-195 mimic reagent were transfected into miR-195 mimic+vector group and miR-195 mimic+FGF7 group, respectively. The effect of FGF7 on the proliferation of miR-195 mimic SCC-4 cells was detected by MTS assay. The expression of phosphorylated phosphatidylinositol 3-kinase(pPI3 K) and phosphorylated protein kinase B(pAKT) protein was detected by Western blotting. Results The cell proliferation ability of the miR-195 mimic group was lower than that of the control group on the 3 rd and 4 th day of cell attachment(P<0.05). Bioinformatics software prediction and luciferase reporter gene experiments verified that FGF7 was a target gene of miR-195. On the 2 nd, 3 rd and 4 th day of cell attachment, the proliferation of the miR-195 mimic+FGF7 group was higher than that of the miR-195 mimic+vector group(P<0.05). The relative expression of pI3 K and pAKT protein in the miR-195 mimic group was lower than that in the control group(both P<0.05). The relative expression of pI3 K and pAKT protein in the miR-195 mimic+FGF7 group was higher than that in the miR-195 mimic group(both P<0.05).Conclusion Overexpression of miR-195 can inhibit the proliferation of OSCC cells, which may be due to the down-regulation of FGF7 expression and thus reducing the activity of PI3 K/AKT signaling pathway.更多还原