siAKAP12对宫颈癌细胞增殖和凋亡的影响及机制

Effects of siAKAP12 on the proliferation and apoptosis of cervix cancer cells and its mechanism

目的:探讨常氧和乏氧环境下下调AKAP12表达对人宫颈癌细胞增殖及凋亡的影响,并探索其可能作用机制。方法:常氧和乏氧环境下体外培养人宫颈癌CaSki细胞并转染siRNA阴性片段、siAKAP12特异性片段,应用CCK-8和流式细胞术检测细胞的增殖活性、凋亡率,应用Real-time PCR、Western blot法检测各组细胞中AKAP12、HIF-1α、CAⅨmRNA和蛋白的表达。结果:在常氧及乏氧状态下,siAKAP12后,CaSki细胞增殖能力均明显增强(P<0.05),但乏氧下细胞增殖能力低于常氧状态(P<0.05)。乏氧相对常氧,CaSki细胞凋亡增加(P<0.01),siAKAP12后,CaSki细胞凋亡率却明显降低(P<0.05);AKAP12相同处理情况下,HIF-1α、CAⅨmRNA表达水平均呈上升趋势(P<0.05);siAKAP12后AKAP12蛋白水平显著下调,而HIF-1α、CAⅨ蛋白均呈上调表达,但两者乏氧下低于常氧表达水平(P<0.05)。结论:常氧和乏氧环境下,抑制AKAP12基因可上调CaSki细胞增殖能力,并降低其凋亡,其作用可能是通过调控HIF-1α-CAⅨ信号通路实现的。

Objective:To investigate the effects of down-regulated AKAP12 expression on proliferation and apoptosis of human cervical cancer cells in normoxic and hypoxic enviroments and the possible mechanism.Methods:Human cervical cancer CaSki cells were transfected with negative siRNA and AKAP12 siRNA and cultured in normoxic and hypoxic states.The proliferation activity and apoptosis rate were detected by CCK-8 assay and flow cytometry,respectively.The mRNA and protein expressions of AKAP12,HIF-1αand CAⅨin each group were detected by Real-time PCR and Western blot.Results:In both normoxic and hypoxic environments,the proliferation activity of siAKAP12 CaSki cells increased significantly(P<0.05),while the proliferation rate in the hypoxic state significantly decreased compared with that in the normoxic state(P<0.05).However,the apoptosis of CaSki cells with normal expression of AKAP12 increased in the hypoxia state than that in the normoxic state(P<0.05),and the apoptosis rate significantly decreased after siRNA interference(P<0.05).In the hypoxic state,the expression levels of HIF-1αand CAⅨmRNA showed an increasing trend compared with normoxic state(P<0.05).After siRNA interfered with AKAP12 expression,AKAP12 protein level was significantly decreased and the protein levels of both HIF-1αand CAⅨwere up-regulated,but were lower in the hypoxic state than in the normoxic state(P<0.05).Conclusion:In normoxic and hypoxic environments,inhibition of AKAP12 promoted the proliferation ability of CaSki cells and reduced their apoptosis,and the effect may be realized by regulating HIF-1α-CAⅨsignaling pathway.