黏附分子CD146对Vorinostat诱导卵巢癌细胞凋亡作用的影响

目的:探讨黏附分子CD146对组蛋白去乙酰化酶抑制剂Vorinostat诱导卵巢癌细胞凋亡作用的影响。方法:通过实时聚合酶链反应(real-time PCR)和蛋白质印迹(western blotting)法检测卵巢癌细胞A2780和SKOV3中CD146在组蛋白去乙酰化酶抑制剂Vorinostat作用下的变化情况。通过流式细胞仪(FACS)及细胞克隆形成实验检测CD146单克隆抗体AA98对Vorinostat诱导卵巢癌细胞凋亡和抑制克隆形成的影响,金氏公式法分析联合用药效果。通过Western blotting法比较Vorinostat单药及AA98与Vorinostat联用对卵巢癌细胞中蛋白激酶B/哺乳动物雷帕霉素靶蛋白(AKT/m TOR)信号通路及下游分子的影响。结果:在卵巢癌细胞中黏附分子CD146在Vorinostat作用下被显著性诱导表达,高表达的CD146可能与卵巢癌化疗敏感性有关。CD146单克隆抗体AA98与Vorinostat联用显著增加了卵巢癌细胞的凋亡率,降低了细胞克隆形成能力,差异有统计学意义(P0.05)。金氏公式计算表明两药具有协同效应。AA98可明显降低Vorinostat引起的AKT/m TOR通路活化作用,降低卵巢癌细胞中磷酸化AKT(p-AKT)、磷酸化真核翻译起始因子4E结合蛋白1(p-4E-BP1)及磷酸化核糖体40S小亚基S6蛋白激酶(p-S6K1)的蛋白表达水平(P0.05)。结论:Vorinostat使CD146显著上调,从而降低了卵巢癌化疗敏感性,CD146单抗可能通过逆转和抑制Vorinostat引起的AKT/m TOR通路活化作用,Vorinostat联合CD146单抗对卵巢癌细胞具有明显的协同杀伤效力。

Effect of Adhesion Molecular CD146 on Apoptosis of Ovarian Cancer Cell Induced by Vorinostat

Objective:To investigate the effect of adhesion molecular CD146 expression on apoptosis of ovarian cancer cells induced by Vorinostat. Methods:CD146 expression levels in A2780 and SKOV3 cells were detected after Vorinostat treatment by Real-time PCR and Western blot. The effect of CD146 mAb AA98 on cell apoptosis and colony-forming ability under Vorinostat treatment by FACS and Soft agar colony-forming assay. The synergistic effect of Vorinostat and AA98 was analyzed by gold formula method. Comparison of effect on AKT/mTOR signaling pathway and the downstream molecular between ovarian cancer cells treated by Vorinostat alone or plus with AA98. Results:CD146 was significantly induced by Vorinostat in ovarian cancer cells. Upregulation of CD146 is closely related to chemosensitivity. Combined Vorinostat and AA98 significantly improved cell apoptotic rate and ablated cancer colony formation. The difference was statistically significant ( P<0.05). The synergistic effect after treatment with a combination of Vorinostat with AA98 was proved by gold formula method. AA98 could reduce the activation of AKT/mTOR signaling pathway caused by Vorinostat and could decrease the p-AKT ,p-4E-BP1,p-S6K1 protein levels in ovarian cancer cells (P<0.05). Conclusions:Vorinostat significantly induced the expression of CD146, which might decrease the chemosensitivity of ovarian cancer cells. CD146 mAb may reverse and inhibit the activation of AKT/mTOR signaling pathway caused by Vorinostat and substantially enhance killing of ovarian cancer cells. The two drugs have obvious synergistic antitumor effects.