人乳头瘤病毒HPV31型宫颈癌细胞模型的建立

宫颈癌与人乳头瘤病毒感染密切相关,建立宫颈癌实验模型可为宫颈癌的研究提供理想的模拟实验条件。我们通过应用基因重组技术,分别以HPV31型E6和E7基因为目的基因,通过原核表达、蛋白纯化和免疫小鼠等获得其特异性检测抗体。我们还通过构建E6和E7基因真核表达载体、转染C33A细胞、博莱霉素抗性筛选和表达检测等步骤,获得一种稳定的体外宫颈癌细胞系。经酶切鉴定及测序证实细胞基因组已重组插入质粒中的目的基因。我们已成功筛选到稳定的目的mRNA和蛋白表达的阳性细胞系,建立了稳定的人乳头状瘤病毒31型(HPV31)的宫颈癌细胞株,为研究宫颈癌提供了体外实验模型。

Establishment of the Human Papillomavirus Type 31 Positive Cervical Cancer Cell Line

The establishment of in vitro model will provide optimal conditions for the study of human papillomavirus(HPV)-associated cervical cancer.In this study,E6 and E7 gens of HPV31 were cloned and expressed in E.coli.The recombinant proteins were purified and used as antigens to immunize mice for the production of polyclonal antibody.Mammalian expression plasmid pBudCE4.1-HPV31-E6/E7 was also constructed and transfected into C33A cells.The transfected cells were then selected by Zeocin.The expressions of the E6 and E7 mRNAs and proteins were detected by RT-PCR and Western blot respectively.A stable cervical cancer cell line was established as an in vitro model for the study of human papillomavirus type 31(HPV31) associated cervical cancer.