Abstract：
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.
摘要：
E6蛋白是宫颈癌中已知的癌基因，通过降低肿瘤抑制蛋白P53的表达水平并最终导致细胞增殖增强和凋亡减少，在宫颈癌的发展和进展中起关键作用。因此,抑制E6癌蛋白表达的抗病毒剂有望成为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可以有效且特异性地敲除人乳头瘤病毒18E6mRNA，下调E6蛋白的表达水平，并恢复抑癌基因P53蛋白的表达，从而抑制宫颈癌细胞的生长并增加其凋亡，E6-2、E6-3和E6-5组细胞凋亡率为25.4%,22.4%,和22.2%的HeLa细胞。此外，CRISPR/Cas13a增强顺铂在宫颈癌HeLa细胞中的增殖抑制和凋亡诱导。针对HPVE6mRNA的CRISPR/Cas13a系统可能是治疗人乳头瘤病毒相关宫颈癌的有希望的治疗方法。
