文章：

CRISPR/Cas9系统在宫颈癌发生中的应用

The application of CRISPR/Cas9 system in cervical carcinogenesis 

原文发布日期：2021-08-04 

英文摘要：

Integration of high-risk HPV genomes into cellular chromatin has been confirmed to promote cervical carcinogenesis, with HPV16 being the most prevalent high-risk type. Herein, we evaluated the therapeutic effect of the CRISPR/Cas9 system in cervical carcinogenesis, especially for cervical precancerous lesions. In cervical cancer/pre-cancer cell lines, we transfected the HPV16 E7 targeted CRISPR/Cas9, TALEN, ZFN plasmids, respectively. Compared to previous established ZFN and TALEN systems, CRISPR/Cas9 has shown comparable efficiency and specificity in inhibiting cell growth and colony formation and inducing apoptosis in cervical cancer/pre-cancer cell lines, which seemed to be more pronounced in the S12 cell line derived from the low-grade cervical lesion. Furthermore, in xenograft formation assays, CRISPR/Cas9 inhibited tumor formation of the S12 cell line in vivo and affected the corresponding protein expression. In the K14-HPV16 transgenic mice model of HPV-driven spontaneous cervical carcinogenesis, cervical application of CRISPR/Cas9 treatment caused mutations of the E7 gene and restored the expression of RB, E2F1, and CDK2, thereby reversing the cervical carcinogenesis phenotype. In this study, we have demonstrated that CRISPR/Cas9 targeting HPV16 E7 could effectively revert the HPV-related cervical carcinogenesis in vitro, as well as in K14-HPV16 transgenic mice, which has shown great potential in clinical treatment for cervical precancerous lesions. 

摘要翻译： 

高危型HPV基因组整合至细胞染色质已被证实会促进宫颈癌发生，其中HPV16是最常见的高风险亚型。本研究评估了CRISPR/Cas9系统在宫颈癌发生过程中的治疗效果，尤其针对宫颈癌前病变。我们在宫颈癌/癌前细胞系中分别转染了靶向HPV16 E7的CRISPR/Cas9、TALEN和ZFN质粒。与既往建立的ZFN和TALEN系统相比，CRISPR/Cas9在抑制宫颈癌/癌前细胞系生长、克隆形成及诱导凋亡方面展现出相当的效率和特异性，这种效应在源自低度宫颈病变的S12细胞系中尤为显著。此外，在异种移植瘤实验中，CRISPR/Cas9有效抑制了S12细胞系的体内成瘤能力并影响相关蛋白表达。在HPV驱动的自发性宫颈癌K14-HPV16转基因小鼠模型中，宫颈局部施用CRISPR/Cas9治疗可引起E7基因突变，恢复RB、E2F1和CDK2的表达，从而逆转宫颈癌表型。本研究证实：靶向HPV16 E7的CRISPR/Cas9能有效逆转HPV相关宫颈癌变进程，这一效应在体外实验和K14-HPV16转基因小鼠模型中均得到验证，展现出治疗宫颈癌前病变的巨大临床潜力。

原文链接：

The application of CRISPR/Cas9 system in cervical carcinogenesis