人乳头瘤病毒16型亚基因DNA体外转化功能的细胞学研究

利用HZIP16和HZIP16K(见材料和方法)质粒,将人乳头瘤病毒16型(HPV-16)的全早期区基因及其开放读码框架(ORF)E6-E7分别转入ψ2细胞,所产生的重组病毒能诱导NIH3T3细胞发生转化。转化细胞具有恶性细胞的生物学和形态学特征,可在0.3％软琼脂中形成集落,可使裸鼠致瘤。Southern blot证明,HPV-16 E6-E7 ORFs序列以整合形式存在于转化细胞和裸鼠肿瘤细胞DNA中,表明HPV-16 DNA具有体外诱导NIH3T3细胞恶性转化的作用,E6-E7 ORFs是诱导细胞转化的关键基因。

TRANSFORMING ACTIVITY OF HUMAN PAPILLOMAVIRUS TYPE 16 SUBGENOMIC DNA IN VITRO

There are increasing evidences that human papillomavirus type 16(HP-V-16)is a most prevalent virus associated with human uterine cervical cancer. In order to prove the transforming potentiality of HPV-16 and identify the relevant transforming gene region of this virus, the NIH3T3 cells were transfected with the total early gene or E6-E7 ORFs of HPV-16 by using the recombinant retrovirus. The transformed NIH3T3 cells appeared morphological characteristic of malignant cells. The polyclonal transformed cells shown anchorage independent growth in 0.3% soft agar. Tumors were developed in all nude mice in 2 weeks after subcutaneous incula-tion of transformed cells and the tumors were diagnosed histologically as fibroscarcoma with low differentiation. Southern blot showed that HpV-16 E6-E7 ORFs seqence were integrated into the chromosomal DNA of transformed cells and cancers produced in nude mice. These results shown that HPV-16 DNA have the transforming potentiality in NIH3T3 cells and E6-E7 ORFs may be the transforming genes of the virus.The transformation of NIH3T3 cells of our study provides a model for analyzing the functions of HPV-16 DNA in the genesis of the human cervical carcinoma.