近二十年来越来越多的证据表明,人乳头瘤病毒16型(HPV-16)与宫颈癌有密切关系.病毒体外转化细胞功能的研究成为确定HPV-16是否为宫颈癌病因的重要依据. 本实验先用HPV-16全基因体外诱导NIH3T3细胞转化.含有HPV-16全基因的重组质粒pSV2neo/HPV-16,通过磷酸钙沉淀方法转染NIH3T3细胞.转染后细胞经过4～6周选择培养,筛选出转化细胞克隆.任意挑选5个克隆扩大培养成系,转化细胞在形态上变圆,折光性增强,生长速度快,接触抑制消失.核酸杂交显示HPV-16DNA以多位点整合形式存在于所有5个单克隆转化细胞系中,并且病毒基因的整 More and more evidences over the past two decades indicate that human papillomavirus type 16 (HPV-16) is closely related to cervical cancer and that the study of the function of the virus in vitro transformed cells is an important basis for determining whether HPV-16 is the cause of cervical cancer In this experiment, NIH3T3 cells were induced by in vitro transduction of all HPV-16 genes, and the recombinant plasmid pSV2neo / HPV-16 containing the entire HPV-16 gene was transfected into NIH3T3 cells by calcium phosphate precipitation Select the culture, filter out the transformed cell clones.Any selection of 5 clones expanded culture lines, the transformed cells in the morphological round, enhanced refractive index, growth rate, contact inhibition disappear.Nucleic acid hybridization showed that HPV-16DNA multi-site integration The form is present in all 5 monoclonal transformed cell lines and the entire viral gene