Department of Epidemiology and Health Statistics, College of Public Health, Zhengzhou University, No. 100 Kexue Avenue, Zhengzhou, Henan 450001, People's Republic of China.
Department of Epidemiology and Health Statistics, College of Public Health, Zhengzhou University, No. 100 Kexue Avenue, Zhengzhou, Henan 450001, People's Republic of China.
Gene. 2024 Aug 30;921:148527. doi: 10.1016/j.gene.2024.148527. Epub 2024 May 6.
The E6 protein is a known oncogene in cervical cancer and plays a key role in the development and progression of cervical cancer by reducing the expression level of the tumor suppressor protein P53 and ultimately leading to enhanced cell proliferation and reduced apoptosis. Therefore, antiviral agents that inhibit the expression of E6 oncoprotein are expected to be potential therapies for human cervical cancer. Here we developed CRISPR/Cas13a: crRNA dual plasmid system and demonstrated that CRISPR/Cas13a could effectively and specifically knock down human papillomavirus 18 E6 mRNA, downregulate the expression level of E6 protein, and restore the expression of the tumor suppressor gene P53 protein, thereby inhibiting the growth of cervical cancer cells and increasing their apoptosis, the E6-2, E6-3, and E6-5 groups resulted in apoptosis rates of 25.4%, 22.4%, and 22.2% in HeLa cells. Moreover, CRISPR/Cas13a enhances the proliferation inhibition and apoptosis induction of cisplatin in cervical cancer HeLa cells. The CRISPR/Cas13a system targeting HPV E6 mRNA may be a promising therapeutic approach for the treatment of human papillomavirus-associated cervical cancer.
E6 蛋白是宫颈癌中的一种已知致癌基因，通过降低肿瘤抑制蛋白 P53 的表达水平，在宫颈癌的发生和发展中发挥关键作用，最终导致细胞增殖增强和细胞凋亡减少。因此，抑制 E6 癌蛋白表达的抗病毒药物有望成为治疗人类宫颈癌的潜在疗法。在这里，我们开发了 CRISPR/Cas13a：crRNA 双质粒系统，并证明 CRISPR/Cas13a 可以有效地特异性敲低人乳头瘤病毒 18 E6 mRNA，下调 E6 蛋白的表达水平，并恢复肿瘤抑制基因 P53 蛋白的表达，从而抑制宫颈癌细胞的生长并增加其细胞凋亡，E6-2、E6-3 和 E6-5 组导致 HeLa 细胞的凋亡率分别为 25.4%、22.4%和 22.2%。此外，CRISPR/Cas13a 增强了顺铂对宫颈癌 HeLa 细胞的增殖抑制和凋亡诱导作用。靶向 HPV E6 mRNA 的 CRISPR/Cas13a 系统可能是治疗人乳头瘤病毒相关宫颈癌的一种有前途的治疗方法。
