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[异质性核糖核蛋白E1对人乳头瘤病毒16型早期基因E2、E6表达及宫颈癌细胞生物学功能的影响]

[Effects of hnRNP E1 on expression of early genes E2, E6 of HPV16 and biological function in cervical cancer cells].

作者信息

Song L, Ding L, Feng M J, Li X X, Gao W, Qi Z, Liu H, Wang M, Wang J T

机构信息

Department of Epidemiology, School of Public Health, Shanxi Medical University, Taiyuan 030001, China.

出版信息

Zhonghua Liu Xing Bing Xue Za Zhi. 2021 Feb 10;42(2):321-326. doi: 10.3760/cma.j.cn112338-20191009-00723.

Abstract

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. The cell experiments 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. 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 (<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 (<0.05). Moreover, the late apoptotic rate and the total apoptotic rate increased (<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 (<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 (=0.427), and no E2 protein of HPV16 was detected. 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.

摘要

探讨异质性核糖核蛋白E1(hnRNP E1)对人乳头瘤病毒16型(HPV16)早期基因E2、E6表达及宫颈癌SiHa细胞系生物学功能的影响。细胞实验在宫颈癌SiHa细胞系中进行。上调hnRNP E1前后,分别采用实时荧光定量聚合酶链反应(Real-time PCR)和蛋白质免疫印迹法(Western blot)检测HPV16的E2、E6 mRNA及蛋白表达水平。同时,采用细胞计数试剂盒-8(CCK-8)法和流式细胞术评估细胞增殖、周期及凋亡情况。数据分析采用SPSS 22.0和Graphpad Prism 7.0软件。与空白组和空质粒组相比,上调hnRNP E1后24、48和72小时细胞活性及增殖能力降低(<0.05),而G0/G1期细胞百分比增加,S期和G2/M期细胞百分比及增殖指数降低(<0.05)。此外,晚期凋亡率和总凋亡率升高(<0.05)。hnRNP E1上调组中HPV16的E6 mRNA和蛋白表达水平显著低于空白组和空质粒组,差异有统计学意义(<0.05),呈现随着HPV16 E6表达降低细胞增殖指数下降和总凋亡率升高的趋势。三组间HPV16的E2 mRNA表达无显著差异(=0.427),且未检测到HPV16的E2蛋白。hnRNP E1可抑制HPV16 E6癌基因的转录和翻译,进而抑制宫颈癌细胞增殖并促进其凋亡,提示hnRNP E1可能是预防宫颈病变的潜在靶向标志物。但在SiHa细胞中未发现hnRNP E1与HPV16 E2之间存在关联。

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