人乳头瘤病毒16型基因组体外转化活性的研究

Research on transformation of NIH3T3 cells in vitro with human papillomavirus type 16 genome

目的应用重组的人乳头瘤病毒16型（HPV－16）基因组体外转化NIH／3T3细胞，以评价HPV－16的转化活性。方法将HPV－16基因组正向括人pSV2／neo质粒，构建成pSV2－neo／HPV－16真核细胞表达质粒；用磷酸钙转染法，将其导人体外培养的NIH／3T3细胞；对经转化的细胞进行G418选择培养和核酸杂交分析。结果经选择培养获得了具有恶性表型的转化细胞，其主要表现为接触性抑制消失和非锚地依赖性生长；核酸杂交证明，转化细胞内含有HPV—16DNA序列。结论HPV—16基因组具有体外转化NIH/3T3细胞的活性。

To evaluate transforming activity from human papillomavirus type 16 (HPV-16) bytransformation of NIH/3T3 cells in vitro with recombinant HPV-16 genome. methods A eukeqoticexpression plasmid (pSV2-neo/HPV - 16) was costructed by insexting HPV - 16 genome into PSV/neo vectorin sense orientation. The expression plasmid was clrawn into NIH/3T3 cells by using calcium phosphatetansfection. Results The transformed cells showed disappearance of contact inhibihon and anchorageindependent growth. Dot blot analysis of DNA from the transformed cells indicated the transformed cellscontained HPV - 16 DNA sequence. Condusions HPV-16 genome can transform NIH/3T3 cells in vitro.