帕瑞昔布经miR-152/ERBB3信号通路对卵巢癌Skov3细胞的增殖抑制和凋亡诱导作用

Effect of Parecoxib on Proliferation Inhibition and Apoptosis Induction of Ovarian Cancer Skov3 Cells via the miR-152/ERBB3 Signaling Pathway

目的探讨帕瑞昔布经miR-152/表皮生长因子受体3(ERBB3)信号通路对卵巢癌Skov3细胞增殖的抑制和凋亡诱导作用。方法体外培养Skov3细胞,阴性对照组加入无血清DMEM培养基,帕瑞昔布低、中、高剂量组分别加入终浓度为50,100,200μmol/L的帕瑞昔布,阳性对照组加入含有终浓度为4μmol/L的顺铂,48 h后以四噻唑蓝(MTT)比色法检测Skov3细胞增殖情况,以流式细胞仪检测细胞凋亡情况,同时以实时荧光定量聚合酶链式反应(RT-PCR)检测Skov3细胞中miR-152,ERBB3,以及环氧合酶2(COX-2)、磷脂酰肌醇3激酶(PI3K)、B细胞淋巴瘤/白血病-2 (Bcl-2)、Bcl-2相关X蛋白(Bax)和半胱氨酸的天冬氨酸蛋白水解酶3(Caspase-3)信使核糖核酸(mRNA)水平。结果与阴性对照组相比,帕瑞昔布低、中、高剂量组和阳性对照组Skov3细胞增殖率均显著降低,Skov3细胞凋亡率均显著增加(P <0.05);其miR-152,ERBB3,COX-2,PI3K,Bcl-2 mRNA水平均显著降低,Bax和Caspase-3mRNA水平均显著升高(P <0.05)。且帕瑞昔布低、中、高剂量组上述指标变化均呈剂量依赖性(P <0.05);与帕瑞昔布高、中、低剂量组相比,阳性对照组上述指标变化更明显(P <0.05)。结论帕瑞昔布可能通过抑制miR-152/ERBB3信号通路,从而抑制Skov3细胞增殖和诱导其凋亡。

Objective To investigate the effect of parecoxib on proliferation inhibition and apoptosis induction of ovarian cancer Skov3 cells via the miR-152/ERBB3 signaling pathway.Methods Skov3 cells were cultured in vitro,and serum-free DMEM medium was added to the negative control group,parecoxib with final concentrations of 50,100,and 200μmol/L was added to the low-dose,medium-dose and high-dose parecoxib groups,respectively,while cisplatin with final concentration of 4μmol/L was added to the positive control group.After 48 h,the proliferation of Skov3 cells were detected by methyl thiazolyl tetrazolium(MTT)assay,the apoptosis of Skov3 cells were dected by flow cytometry,and the levels of miR-152,ERBB3,cyclooxygenase-2(COX-2),phosphatidylinositol 3 kinase(PI3 K),B cell lymphoma/leukemia-2(Bcl-2),Bcl-2 associated X protein(Bax)and Caspase-3 mRNA in Skov3 cells were detected by RT-PCR method.Results Compared with those in the negative control group,the proliferation rate of Skov3 cells was significantly decreased,and the apoptosis rate of Skov3 cells was significantly increased(P<0.05),the levels of miR-152,ERBB3,COX-2,PI3 K and BCL-2 mRNA in Skov3 cells were significantly decreased,the levels of Bax and Caspase-3 mRNA in Skov3 cells were significantly increased in low-dose,medium-dose and high-dose parecoxib groups and positive control group(P<0.05),and the effect was dose-dependent in low-dose,medium-dose and high-dose parecoxib groups(P<0.05).Compared with those in low-dose,medium-dose and high-dose parecoxib groups,the changes of the above indexes in the positive control group were more obvious(P<0.05).Conclusion Parecoxib can inhibit the proliferation of Skov3 cells and induce the apoptosis of Skov3 cells by inhibiting the miR-152/ERBB3 signaling pathway.