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目的:探讨人乳头瘤病毒(human papillomavirus,HPV)16E6对宫颈癌细胞中高迁移率族蛋白B1(high mobility group box-B1,HMGB1)主动释放及细胞侵袭能力的影响。方法:构建HPV16E6表达质粒,并转染至宫颈癌C33A细胞,FCM法分析细胞存活率;半定量RT-PCR法检测转染24h前后C33A细胞中HPV16E6基因的表达水平;蛋白质印迹法检测转染前后C33A细胞的细胞核、细胞质以及细胞培养液中HMGB1蛋白水平的变化情况;Transwell实验检测细胞侵袭能力的变化。结果:HPV16E6基因转染C33A细胞后,细胞核中HMGB1蛋白水平明显降低(P<0.01),而细胞质及细胞培养液中HMGB1蛋白水平均明显升高(P<0.01)。转染后细胞存活率未见明显差异,但细胞穿膜个数明显多于未转染组(P<0.01)。结论:HPV16E6可诱导宫颈癌细胞内HMGB1的主动释放,提高肿瘤细胞的侵袭能力。
Objective: To investigate the effect of human papillomavirus (HPV) 16E6 on the active release of high mobility group box-1 (HMGB1) and cell invasion in cervical cancer cells. Methods: The HPV16E6 expression plasmid was constructed and transfected into C33A cells. The cell viability was analyzed by FCM. The expression of HPV16E6 gene in C33A cells was detected by semi-quantitative RT-PCR. The expression of HPV16E6 gene was detected by Western blotting before and after transfection C33A cells in the nucleus, cytoplasm and cell culture medium HMGB1 protein levels changes; Transwell assay to detect changes in cell invasiveness. Results: The level of HMGB1 protein in the nucleus of C33A cells transfected with HPV16E6 gene was significantly decreased (P <0.01), while the levels of HMGB1 protein in cytoplasm and cell culture medium were significantly increased (P <0.01). There was no significant difference in cell viability after transfection, but the number of cell transplants was significantly more than that of untransfected cells (P <0.01). Conclusion: HPV16E6 can induce the active release of HMGB1 in cervical cancer cells and increase the invasion ability of tumor cells.