论文部分内容阅读
目的:探讨hnRNP E1对HPV16早期基因E2、E6表达的调控及宫颈癌细胞生物学功能的影响。方法:采用体外细胞实验方法，对HPV16阳性人宫颈鳞癌SiHa细胞株采用hnRNP E1cDNA质粒上调hnRNP E1表达，于上调前、后采用Real-time PCR和Western blot分别检测各组细胞HPV16 E2、E6 mRNA和蛋白表达水平，同时，应用CCK-8法和流式细胞术检测细胞增殖、周期及凋亡。采用SPSS 22.0和Graphpad Prism 7.0软件进行相关资料分析。结果:随着转染时间的增加（24、48、72 h），hnRNP E1过表达组的SiHa细胞活性及处于增殖状态的细胞数逐渐减少（n P<0.05），而G0/G1期细胞比例增高，S、G2/M期细胞比例及增殖指数降低（n P<0.05），晚期凋亡率和细胞总凋亡率逐渐增加（n P<0.05）。hnRNP E1过表达组HPV16 E6 mRNA和蛋白表达水平低于空白组和空质粒组，差异有统计学意义（n P<0.05），且随着HPV16 E6 mRNA和蛋白表达水平的降低，SiHa细胞增殖指数下降，而总凋亡率有增加趋势。3组间HPV16 E2 mRNA表达量差异无统计学意义（n P=0.427），HPV16 E2蛋白未被检测到。n 结论:hnRNP E1可抑制HPV16 E6的转录和翻译，进而抑制宫颈癌细胞增殖，促使凋亡，hnRNP E1可作为抑制宫颈癌变的潜在靶标。但本研究未发现hnRNP E1与HPV16 E2在SiHa细胞中的关系。“,”Objective:To explore the effects of hnRNP E1 on the expression of early genes E2, E6 of HPV16 and the biological function in cervical cancer SiHa cell lines.Methods:The cell experiments n in vitro were carried out in cervical cancer cell lines SiHa. The expression levels of E2, E6 mRNA and protein of HPV16 were detected by Real-time PCR and Western blot, respectively, before and after up-regulating hnRNP E1. Meanwhile, the cell proliferation, cycle and apoptosis were evaluated by CCK-8 and flow cytometry. Data analyses were performed using SPSS 22.0 and Graphpad Prism 7.0 software.n Results:Compared with the blank and the blank plasmid group, the cells activity and proliferation decreased at 24, 48 and 72 h after up-regulating hnRNP E1 (n P<0.05), while the percentage of cells in G0/G1 phase increased and the percentage in S and G2/M phase and proliferation index decreased (n P<0.05). Moreover, the late apoptotic rate and the total apoptotic rate increased (n P<0.05). The expression levels of E6 mRNA and protein of HPV16 in hnRNP E1 up-regulated group were significantly lower than that in both blank group and blank plasmid group, the differences were significant (n P<0.05), showing the tendency of cells proliferation index decrease and total apoptotic rate increase with decreased HPV16 E6 expression. There were no significant differences in the expression of E2 mRNA of HPV16 among the three groups (n P=0.427), and no E2 protein of HPV16 was detected.n Conclusions:hnRNP E1 could inhibit the transcription and translation of E6 oncogene of HPV16 and further inhibit the proliferation and promote apoptosis of cervical cancer cells, suggesting that hnRNP E1 might be a potential target marker to prevent cervical lesions. But no association between hnRNP E1 and HPV16 E2 was found in SiHa cells.