Zhu Yingping, Yang Wenjuan, Zhuang Yulong, Wang Feifei, Ge Yanlu, Jiang Jun, Feng Danping
Department of Obstetrics and Gynecology, The First Affiliated Hospital of Zhejiang Chinese Medical University (Zhejiang Provincial Hospital of Chinese Medicine), P.R. China.
First School of Clinical Medicine, Zhejiang Chinese Medical University, P.R. China.
J Microbiol Biotechnol. 2024 Dec 28;34(12):2484-2491. doi: 10.4014/jmb.2408.08041. Epub 2024 Oct 30.
Current cancer burden caused by persistent infection with human papillomaviruse genotype 16 (HPV16) cannot be ignored. The related mechanisms of oncoproteins E6 and E7 from HPV16 on keratinocyte malignant transformation need to be further elucidated. GSE3292 dataset analysis revealed the upregulation of ETS transcription factor 3 (ELF3) and cyclin E2 (CCNE2). To verify whether there is an interaction between ELF3 and CCNE2, E74 like ELF3 and CCNE2 expression profiles as well as their putative binding sites were analyzed using bioinformatics. Retroviruses encoding HPV16 E6 and E7 genes were used to induce immortalization of human foreskin keratinocytes (HFKs) in vitro. Dual luciferase reporters assay was used to verify the binding of ELF3 and CCNE2. The effect of ELF3 on the immortalized cells was investigated using CCK-8 assay, cell cycle analysis and western blot. ELF3 and CCNE2 presented overexpression patterns in head and neck squamous cell carcinoma. HPV16 E6/E7-expressing HFKs showed enhanced viability, accelerated cell cycle as well as upregulated ELF3 and CCNE2. ELF3 overexpression enhanced the activity of CCNE2 promoter. ELF3 silencing reduced viability, induced cell cycle arrest and suppressed expressions of CCNE2, E6 and E7 in HPV16 E6/E7-expressing HFKs. Downregulation of ELF3 played an inhibiting role in the malignant transformation of HPV16 E6/E7-immortalized HFKs by decreasing CCNE2 expression.
由人乳头瘤病毒16型（HPV16）持续感染引起的当前癌症负担不容忽视。HPV16的癌蛋白E6和E7对角质形成细胞恶性转化的相关机制需要进一步阐明。GSE3292数据集分析显示ETS转录因子3（ELF3）和细胞周期蛋白E2（CCNE2）上调。为了验证ELF3和CCNE2之间是否存在相互作用，使用生物信息学分析了E74样ELF3和CCNE2的表达谱及其假定的结合位点。编码HPV16 E6和E7基因的逆转录病毒用于在体外诱导人包皮角质形成细胞（HFK）永生化。双荧光素酶报告基因检测用于验证ELF3和CCNE2的结合。使用CCK-8检测、细胞周期分析和蛋白质印迹研究ELF3对永生化细胞的影响。ELF3和CCNE2在头颈部鳞状细胞癌中呈现过表达模式。表达HPV16 E6/E7的HFK显示活力增强、细胞周期加速以及ELF3和CCNE2上调。ELF3过表达增强了CCNE2启动子的活性。ELF3沉默降低了活力，诱导细胞周期停滞，并抑制了表达HPV16 E6/E7的HFK中CCNE2、E6和E7的表达。ELF3的下调通过降低CCNE2表达在HPV16 E6/E7永生化HFK的恶性转化中发挥抑制作用。
