Cav3.1在宫颈癌组织中的表达及靶向下调Cav3.1对宫颈癌HeLa细胞增殖和凋亡的影响

Expression of Cav3.1 in cervical cancer tissue and the effect of targeted down-regulation of Cav3.1 on the proliferation and apoptosis of cervical cancer HeLa cells

目的:分析Cav3.1在宫颈癌组织中的表达及靶向下调Cav3.1对人宫颈癌HeLa细胞增殖和凋亡的影响。方法:采用免疫组织化学染色法检测Cav3.1在68例宫颈癌组织和40例非癌宫颈组织中的表达;实时定量聚合酶链反应(Real-time PCR)、Western blot检测宫颈癌HeLa细胞中Cav3.1的表达;采用siRNA瞬时转染技术靶向下调Cav3.1表达,CCK-8检测下调Cav3.1表达后宫颈癌HeLa细胞增殖情况,流式细胞仪检测下调Cav3.1表达后宫颈癌HeLa细胞凋亡情况;Western blot检测宫颈癌HeLa细胞中Bcl-2、Bax、Cleaved-Caspase3的表达。结果:Cav3.1在宫颈癌组织中的表达显著高于对照组(P<0.01)。宫颈癌HeLa细胞中Cav3.1的表达水平显著高于HcerEpic细胞(P<0.01)。与对照组相比,siRNA-Cav3.1组的Cav3.1相对表达水平显著下降(P<0.01)。与空白对照组、mock组相比,siRNA-Cav3.1组HeLa细胞的增殖能力减弱,凋亡能力增强,差异有统计学意义(P<0.01)。Western blot显示siRNA-Cav3.1组Bax、Cleaved-Caspase3的相对表达量比对照组升高,而Bcl-2降低(P<0.01)。结论:Cav3.1在宫颈癌组织和细胞中高表达。靶向下调Cav3.1可以抑制HeLa细胞的增殖、促进其凋亡。Cav3.1在宫颈癌的生物学行为中可能起着关键的作用,有望成为一种新的有效的宫颈癌治疗靶点及肿瘤检测的分子生物学标志物。

Objective:To investigate the expression of Cav3.1 in cervical cancer and the effect of targeted down-regulation of Cav3.1 on proliferation and apoptosis of human cervical cancer HeLa cells.Methods:The expression of Cav3.1 in 68 cervical cancer tissues and 40 non-cancerous cervical tissues was detected by immunohistochemical staining.Real-time quantitative polymerase chain reaction and Western blot were used to detect the expression of Cav3.1 in cervical cancer HeLa cells.siRNA transient transfection technique was used to target down-regulation of Cav3.1.Proliferation of cervical cancer HeLa cells after down-regulation of Cav3.1 was detected by CCK-8,and apoptosis of cervical cancer HeLa cells after down-regulation of Cav3.1 was detected by flow cytometry.Western blot was used to detect the expression of Bcl-2,Bax,Cleaved-Caspase3 in cervical cancer HeLa cells.Results:The expression of Cav3.1 in cervical cancer tissue was significantly higher than that in control group(P<0.01).The expression level of Cav3.1 in HeLa cell line was significantly higher than that in HcerEpic cell line(P<0.01).Compared with the control group,the relative expression level of Cav3.1 in siRNA-Cav3.1 group was significantly decreased(P<0.01).Compared with the blank control group and the mock group,the proliferation ability of HeLa cells in the siRNA-Cav3.1 group was weakened,and the apoptosis ability was enhanced,and the difference was statistically significant(P<0.01).Western blot showed that the relative expression levels of Bax and Cleaved-Caspase3 in the siRNA-Cav3.1 group were higher than those in the control group,while Bcl-2 was lower(P<0.01).Conclusion:Cav3.1 is highly expressed in cervical cancer tissues and cells.Targeted down-regulation of Cav3.1 can inhibit proliferation and promote apoptosis of HeLa cells.Cav3.1 may play a key role in the biological behavior of cervical cancer,and is expected to become a new effective therapeutic target for cervical cancer and a molecular biological marker for tumor detection.