Kong Qingqin, Wang Wenfeng, Li Ping
Department of Obstetrics and Gynecology, Linyi People's Hospital Linyi City, Shandong Province, China.
Int J Clin Exp Pathol. 2015 Dec 1;8(12):15808-13. eCollection 2015.
Cervical cancer is the second leading malignant tumor in women. Human papillomavirus 16 (HPV16) is one risk factor for cervical cancer, with its expressed E7 protein can facilitate the transformation of cervical epithelial cells. MicroRNA-21 (miR-21) is one important tumor growth regulatory factor involving in angiogenesis, tumor invasion and metastasis. This study thus aimed to investigate the role of high-risk HPV16 E7 protein in regulating miR-21 expression in cervical carcinoma and its related functions. Hela cells were transfected with pcDNA-HPV16 E7 expressing vectors. The expression level of E7 was determined by Western blotting, while miR-21 level was quantified by real-time PCR. The alternation of tumor cell proliferation is determined by transfecting miR-21 inhibitor into E7-overexpressing Hela cells. Cell apoptosis was studied by caspase-3 assay, while cell invasion was illustrated in Transwell chamber. The overexpression of HPV E7 protein facilitated the expression of miR-21, which potentiated Hela cell proliferation and invasion. The inhibition of miR-21 in E7-overexpressin Hela cells can inhibit both proliferation and invasion, but without significant effects on caspase-3 activity. HPV16 E7 protein can up-regulate host miR-21 expression, thus elevating cervical carcinoma cell growth, proliferation and invasion. Therefore, E7 protein is one critical factor in occurrence and progression of cervical carcinoma.
宫颈癌是女性第二大常见恶性肿瘤。人乳头瘤病毒16型(HPV16)是宫颈癌的一个风险因素,其表达的E7蛋白可促进宫颈上皮细胞的转化。微小RNA-21(miR-21)是一种重要的肿瘤生长调节因子,参与血管生成、肿瘤侵袭和转移。因此,本研究旨在探讨高危型HPV16 E7蛋白在调节宫颈癌中miR-21表达及其相关功能中的作用。用表达载体pcDNA-HPV16 E7转染Hela细胞。通过蛋白质免疫印迹法测定E7的表达水平,同时通过实时聚合酶链反应定量miR-21水平。将miR-21抑制剂转染到过表达E7的Hela细胞中来确定肿瘤细胞增殖的变化。通过半胱天冬酶-3检测研究细胞凋亡,而在Transwell小室中观察细胞侵袭情况。HPV E7蛋白的过表达促进了miR-21的表达,从而增强了Hela细胞的增殖和侵袭能力。在过表达E7的Hela细胞中抑制miR-21可同时抑制增殖和侵袭,但对半胱天冬酶-3活性无显著影响。HPV16 E7蛋白可上调宿主miR-21的表达,从而促进宫颈癌细胞的生长、增殖和侵袭。因此,E7蛋白是宫颈癌发生和发展的一个关键因素。