HPV16 E2基因促进宫颈癌SiHa细胞的凋亡

HPV16 E2 Gene Promotes Apoptosis of Cervical Cancer SiHa Cells

为了探讨HPV16 E2基因对宫颈癌SiHa细胞凋亡的影响,并深入揭示其作用机制,本研究采用脂质体转染p-CAG-MYC-HPV16 E2至宫颈癌SiHa细胞中,MTT法检测各组细胞活力,流式细胞术检测各组细胞凋亡水平,RT-PCR以及Western blotting检测各组凋亡相关基因mRNA以及蛋白表达水平。Western blotting检测NF-κB信号通路的变化。结果显示,p-CAG-MYC-HPV16 E2转染后,细胞活力明显下降,与空载体组相比有显著性差异。转染后SiHa细胞凋亡率明显高于空载体组。此外,凋亡相关蛋白Bax的蛋白表达水平明显增加,Bcl2蛋白表达水平明显减少,表明HPV16 E2基因促进宫颈癌SiHa细胞凋亡。另外,转染p-CAG-MYC-HPV16 E2后,SiHa细胞NF-κB p65蛋白表达显著高于空载体组。HPV16 E2基因促进宫颈癌SiHa细胞凋亡,其机制涉及调控NF-κB通路,以期为宫颈癌的防治提供新思路。

To investigate the role of HPV16 E2 gene in apoptosis of cervical cancer siHa cell and further explore its mechanism,p-CAG-MYC-HPV16 E2 was transferred to SiHa Cell to overexpression of HPV16 E2 gene.The cell viability was measured by MTT assay.The rate of apoptotic cells were analyzed by flow cytometry using Annexin V-FITC/PI staining.The apoptosis-related genes mRNA levels and protein expressions were detected by RT-PCR as well as Western blotting,respectively.The NF-κB signaling was analyzed by Western blotting.The cell viability was gradually decreased in time dependence after p-CAG-MYC-HPV16 E2 transfection compared to vehicle.The apoptosis rate in p-CAG-MYC-HPV16 E2 group was significantly higher than vehicle.In addition,the mRNA level and protein expression of Bax were dramatically increased,while the mRNA level and protein expression of Bcl-2 were decreased after p-CAG-MYC-HPV16 E2 transfection.Furthermore,transfection of p-CAG-MYC-HPV16 E2 significantly upregulated the expression of NF-κB p65.HPV16 E2 promotes apoptosis of SiHa cells via regulating NF-κB signaling pathway,which provided a new perspective for Cervical Cancer treatment.