腺病毒伴随病毒表达载体表达人乳头瘤病毒18型E6E7基因构建及其转化作用的鉴定  

Construction of human papilloma virus type 18 E6E7 genes in adeno associated virus expression vector and checking its activity for malignant transformation

作　　者：岑山[1] 滕智平[1] 张月[1] 沈忠英[2] 许锦阶[2] 杜宾[3] 曾毅[1] 

机构地区：[1]中国预防医学科学院病毒学研究所肿瘤室,北京100052 [2]汕头大学医学院病理教研室 [3]哈佛大学

出　　处：《中华实验和临床病毒学杂志》2003年第1期5-9,共5页Chinese Journal of Experimental and Clinical Virology

摘　　要：目的　探讨人乳头瘤病毒 (HPV)的E6E7基因在细胞恶性转化中所起的作用。方法　将人乳头瘤病毒 (HPV)的E6E7基因克隆至腺病毒伴随病毒表达载体中 ,通过包装的重组病毒感染 ,将E6E7基因导入并整合到永生 2 93细胞的基因组中。结果　本研究成功地构建了HPV18E6E7AAV病毒并感染了永生 2 93细胞 ,PCR Southern杂交分析表明E6E7基因在转化细胞 2 93TL中确有表达 ,转化细胞 2 93TC和 2 93TL具有明显的转化表型 ,和亲本 2 93细胞相比 ,生长速度快 ,接触抑制消失 ,集落形成率提高 2 0倍 ,且集落明显增大 ,形成时间短。结论　成功地构建了HPV18E6E7AAV病毒 ,HPV18E6E7基因可引起永生化人上皮细胞 2 93的恶性转化。此病毒可用于感染正常上皮细胞 。Objective To construct human papillomavirus type 18 (HPV18 E6E7) adeno associated virus (AAV) for studying the role of HPV E6E7 in the development of human cancer. Methods HPV18 E6E7 genes were inserted into adeno associated virus expression vector and then infected 293 cell line.The expression of HPV18 E6E7 genes were confirmed by using RT PCR/Southern blot assay. Results There was HPV18 E6E7 genes in the malignantly transformed cell line. The 293TL cells compared with the parent cells transformed cells grew more rapidly,lost their contact inhibition and formed more and large colonies in soft agar.Conclusion HPV18 E6E7 AAV was successfully constructed and could induce malignant transformation. HPV18 E6E7 AAV can be use for studying the immortalization and malignant transformation of human normal epithelial cells.

关 键 词：鉴定 人乳头状瘤病毒 遗传转化 依赖病毒 基因表达 

分 类 号：R373[医药卫生—病原生物学]