CIRBP对人卵巢癌细胞SKOV3的促癌功能研究

The Cancer-promoting Function of CIRBP on Human Ovarian Cancer Cell SKOV3

目的探索冷诱导RNA结合蛋白(CIRBP)对人卵巢癌细胞SKOV3的促癌作用。方法采用生物信息学分析CIRBP在卵巢癌中的表达情况,建立CIRBP基因沉默和过表达的慢病毒稳转SKOV3细胞株,采用CCK-8法、流式细胞法、细胞划痕法分别检测细胞的增殖、凋亡、周期和迁移,采用细胞免疫荧光法检测β-catenin核转位,采用RT-qPCR检测CIRBP mRNA表达水平,采用Western blot检测CIRBP、β-连环蛋白(β-catenin)和磷酸化β-连环蛋白(p-β-catenin)的表达水平。结果CIRBP在卵巢癌中的表达下调(P<0.05),成功构建CIRBP沉默和过表达细胞模型,CIRBP沉默导致细胞增殖、迁移能力下降(P<0.05),细胞凋亡率、G0/G1期比例上升(P<0.05),β-catenin蛋白核转位水平下降(P<0.05),β-catenin蛋白水平下调(P<0.05),而p-β-catenin蛋白水平上调(P<0.05),CIRBP的过表达则相反。结论CIRBP促进β-catenin的表达和核转位,从而影响卵巢癌的进展。

Objective To explore the cancer-promoting effect of cold inducible RNA binding protein(CIRBP)on human ovarian cancer cell SKOV3.Methods CIRBP expression in ovarian cancer was analyzed using bioinformatics.The SKOV3 cell line stably transfected by lentivirus with CIRBP gene silencing and overexpression was established.CCK-8 was used to detect cell proliferation,flow cytometry was applied to detect cell apoptosis and cell cycle,scratch assay was used to detect cell migration,and cellular immunofluorescence was used to detect the nuclear translocation level ofβ-catenin,CIRBP mRNA expression was detected by RT-qPCR,CIRBP,β-catenin and p-β-catenin protein expression levels were detected by Western blot.Results The expression of CIRBP was significantly downregulated in ovarian cancer(P<0.05).CIRBP silenced and overexpressed cell models were satisfactorily constructed.CIRBP silencing resulted in a significant decrease in cell proliferation(P<0.05),a significant increase in apoptosis(P<0.05),a sizable decrease in migration ability(P<0.05),and the proportion of cells in G0/G1 phase increased,the nuclear translocation level ofβ-catenin decreased(P<0.05),the protein expression ofβ-catenin was down-regulated(P<0.05),while the protein expression of p-β-catenin was significantly up-regulated(P<0.05),and CIRBP over expression was reversed.Conclusion CIRBP promotes the expression and nuclear translocation ofβ-catenin,thereby affecting the progression of ovarian cancer.