人乳头瘤病毒16型E6和E7基因及其突变体转化活性的研究

Transforming Activities of the Mutants of HPV16 E6 and E7 Genes for Balb/c 3T3 Cells

为筛选出可用于研制HPV治疗性疫苗的HPV16型E6和E7基因突变体 ,故将HPV16型原型株 (德国株 )E6和E7基因及其各种突变体分别转染Balb/c3T3细胞 ,观察转染后的细胞在软琼脂培养中的集落形成能力和在裸鼠体内的成瘤能力。结果表明 ,单独转染和共转染HPV16野生型E6和E7基因的Balb/c3T3细胞系 ,在软琼脂中呈集落样生长 ,并在裸鼠体内成瘤 ;而转染E6基因突变体mE6 (5 0G)、E7基因的两种突变体mE7- 1(2 4G2 6G)和mE7- 3(2 4G2 6G6 7R)以及共转染mE6和mE7- 1的Balb/c3T3细胞 ,在软琼脂培养中极少形成集落 ,也不能在裸鼠体内成瘤。提示经结构改造后的HPV16E6和E7基因已失去了对Balb/c3T3细胞的转化活性 ,而保留了免疫原性 ,可用于HPV16相关肿瘤治疗性疫苗的构建。

To select the mutants of HPV type 16 E6 and E7 genes suitable for construction of vaccine for treatment of cervical cancer,the transforming activities of the mutants of HPV16 E6 and E7 genes were studied.Balb/c 3T3 cells were transfected with HPV16 wild and mutated E6 and E7 genes.The G418-resistant clones from each group were examined for their anchorage-independent growth in soft agar and tumorigenicity in nude mice.The results showed that Balb/c 3T3 cell lines transfected with wE6,wE7 and wE6wE7 genes exhibited anchorage-independent growth in soft agar and tumorigenicity in nude mice.Whereas the Balb/c 3T3 cell lines transfeted with mE6(50G),mE7-1(24G26G),mE7-3(24G26G67R) and mE6mE7-1 lost the ability of anchorage-independent growth in soft agar and tumorigenicity in nude mice.The mutants mE6(50G),mE7-1(24G26G) and mE7-3(24G26G67R) could be used as the candidate antigens to develop therapeutic vaccine of HPV16 associated tumors.