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近年来的研究证实,人乳头状瘤病毒16型(HPV16)E6和E7基因表达在人类生殖道肿瘤,特别是宫颈癌的发生及其恶性表型的维持中起着重要作用。我们针对HPV16 E7 mRNA设计的ribozyme经体外实验证明能特异地切割E7 RNA片断,本文利用T7-痘苗病毒表达体系研究了ribozyme在真核细胞内的表达.结果表明,利用痘苗病毒表达系统在CV-1细胞中表达了具有体外切割活性的抗HPV16 E7-ribozyme。同时观察到ribozyme能使细胞内HPV16 E7 RNA的表达水平降低。提示ribozyme在细胞内具有一定的生物学活性,有可能利用ribozyme逆转HPV16阳性的宫颈癌细胞的恶性表型。
Recent studies have confirmed that human papillomavirus type 16 (HPV16) E6 and E7 gene expression plays an important role in the development of human genital tumors, especially the occurrence of cervical cancer and the maintenance of its malignant phenotype. Our ribozyme designed for HPV16 E7 mRNA has been demonstrated in vitro to specifically cleave E7 RNA fragments. In this study, we used T7-vaccinia virus expression system to study the expression of ribozyme in eukaryotic cells. The results showed that the use of vaccinia virus expression system in CV- An anti-HPV16 E7-ribozyme with in vitro cleavage activity was expressed in the cells. At the same time, it was observed that ribozyme can reduce the expression level of HPV16 E7 RNA in cells. It is suggested that ribozyme has a certain biological activity in cells, and it is possible to use ribozyme to reverse the malignant phenotype of HPV16-positive cervical cancer cells.