CHK1-CDK1通路介导的G2/M检验点对卵巢癌细胞增殖和凋亡的影响
Effects of CHK1-CDK1 signaling pathway mediated G2/M checkpoint on proliferation and apoptosis of ovarian cancer cells

目的:探讨检查点激酶1(CHK1)-细胞周期依赖激酶1(CDK1)信号通路异常与上皮性卵巢癌细胞增殖和凋亡的关系。方法:分别将CDK1、CHK1基因特异性shRNA质粒及阴性对照质粒转染卵巢上皮性癌SK-OV-3、OVCAR-3细胞,分别采用MTT法和流式细胞术检测各组细胞增殖和凋亡情况,分别采用RT-PCR和Western blot法检测各组细胞中CDK1和CHK1的表达情况,采用Western blot法检测各组细胞中p-CDK1、CyclinB1、p-CHK1(ser345)、CDC25C、p-CDC25C以及增殖和凋亡相关蛋白PCNA、Ki-67、Caspase8、Cleaved-Caspase3的表达情况。结果:分别沉默CDK1、CHK1基因后,SK-OV-3和OVCAR-3细胞增殖受到抑制、凋亡率增加; PCNA、Ki-67蛋白表达减少,Caspase8、Cleaved-Caspase3蛋白表达增加(P<0.05)。此外,沉默CHK1基因可引起p-CHK1和p-CDC25C表达减少、p-CDK1蛋白表达升高(P<0.05)。结论:CHK1-CDK1信号通路介导的G2/M检验点参与调节卵巢癌细胞的增殖和凋亡。
Aim: To analyze the relationship between CHK1-CDK1 signaling pathway and the proliferation and apoptosis of ovarian cancer cells.Methods: The specific shRNA plasmids and negative control plasmid of CDK1 and CHK1 genes were transfected into ovarian cancer SK-OV-3 and OVCAR-3 cells respectively. The cell proliferation and apoptosis were measured by MTT and FCM assay respectively. The expressions of CDK1 and CHK1 mRNA and protein were detected by RT-PCR and Western blot.The levels of p-CDK1, CyclinB1, p-CHK1, CDC25C, p-CDC25C, PCNA, Ki-67, Caspase8, and Cleaved-Caspase3 proteins were examined by Western blot. The cell proliferation and apoptosis were measured by MTT and FCM assay respectively.Results: As results of CDK1 and CHK1 inhibition by shRNA, the cell proliferation was repressed and cell apoptosis rate was increased in both SK-OV-3 and OVCAR-3 cells, expressions of PCNA and Ki-67 protein were downregulated, and expressions of Caspase8 and Cleaved-Caspase3 were upregulated(P<0.05). Moreover, the levels of p-CHK1 and p-CDC25C were decreased, and p-CDK1 was increased after CHK1 inhibition(P<0.05).Conclusion: G2/M checkpoint mediated by CHK1-CDK1 signaling pathway is implicated in the proliferation and apoptosis regulation of ovarian cancer cells