Hu Zheng, Yu Lan, Zhu Da, Ding Wencheng, Wang Xiaoli, Zhang Changlin, Wang Liming, Jiang Xiaohui, Shen Hui, He Dan, Li Kezhen, Xi Ling, Ma Ding, Wang Hui
Department of Obstetrics and Gynecology, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
Cancer Biology Research Center, Tongji Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan, Hubei 430030, China.
Biomed Res Int. 2014;2014:612823. doi: 10.1155/2014/612823. Epub 2014 Jul 20.
High-risk human papillomavirus (HR-HPV) has been recognized as a major causative agent for cervical cancer. Upon HPV infection, early genes E6 and E7 play important roles in maintaining malignant phenotype of cervical cancer cells. By using clustered regularly interspaced short palindromic repeats- (CRISPR-) associated protein system (CRISPR/Cas system), a widely used genome editing tool in many organisms, to target HPV16-E7 DNA in HPV positive cell lines, we showed for the first time that the HPV16-E7 single-guide RNA (sgRNA) guided CRISPR/Cas system could disrupt HPV16-E7 DNA at specific sites, inducing apoptosis and growth inhibition in HPV positive SiHa and Caski cells, but not in HPV negative C33A and HEK293 cells. Moreover, disruption of E7 DNA directly leads to downregulation of E7 protein and upregulation of tumor suppressor protein pRb. Therefore, our results suggest that HPV16-E7 gRNA guided CRISPR/Cas system might be used as a therapeutic strategy for the treatment of cervical cancer.
高危型人乳头瘤病毒（HR-HPV）已被确认为宫颈癌的主要致病因子。HPV感染后，早期基因E6和E7在维持宫颈癌细胞的恶性表型中发挥重要作用。通过使用成簇规律间隔短回文重复序列（CRISPR）相关蛋白系统（CRISPR/Cas系统，一种在许多生物体中广泛使用的基因组编辑工具）靶向HPV阳性细胞系中的HPV16-E7 DNA，我们首次表明HPV16-E7单向导RNA（sgRNA）引导的CRISPR/Cas系统可在特定位点破坏HPV16-E7 DNA，诱导HPV阳性的SiHa和Caski细胞凋亡并抑制其生长，但对HPV阴性的C33A和HEK293细胞无此作用。此外，E7 DNA的破坏直接导致E7蛋白表达下调和肿瘤抑制蛋白pRb表达上调。因此，我们的结果表明，HPV16-E7 gRNA引导的CRISPR/Cas系统可能用作治疗宫颈癌的一种策略。
