摘要
目的 人乳头瘤病毒 16型 E6 E7区基因编码的蛋白具有转导人上皮细胞 ,使之获得永生化特性的作用。为建立卵巢癌永生细胞系 ,我们对 HPV16 E6 E7转导卵巢癌细胞的方法及转导细胞对顺铂的耐药性进行了研究。 方法 采用脂质体方法 ,将含有 HPV16 E6 E7的重组逆转录病毒载体 p L XSN16 E6 E7,转染逆转录病毒包装细胞 PT6 7,经 G418筛选 ,病毒滴度测定 ,筛选出 1株高滴度 (1.9× 10 7/m l)的产病毒克隆 ,将之转导卵巢癌细胞系SKOV3、3AO,各筛选出 1株稳定表达 HPV16 E6 E7的克隆 ,SKOV3/E6 E7、3AO/E6 E7。经 RT- PCR、免疫细胞化学检测证实 HPV16 E6 E7的表达 ,之后采用 MTT法检测了 HPV16 E6 E7转导的卵巢癌细胞系对顺铂耐药性的变化。结果 HPV16 E6 E7转导的卵巢癌细胞系 SKOV3/E6 E7、3AO/E6 E7对顺铂的耐药性无改变。 结论 HPV16 E6 E7转导的卵巢癌细胞没有发生顺铂耐药性的改变。本研究为采用此法建立卵巢癌永生细胞系 ,特别是建立耐药卵巢癌细胞系奠定了基础。
Objective\ Human papillomavirus type 16 E6E7 region(HPV16E6E7)gene coded protein can transform human epithelial cells,and make them immortalized.In order to establish immortal ovarian carcinoma cells, we established the method of HPV16E6E7 transforming ovarian carcinoma cells,of which the cisplatin\|resistant changes were also studied. Methods\ Using lipofectin method to introduce the recombinant retrovirus plasmid pLXSN16E6E7 containing the HPV16E6E7 into a retrovirus packaging cell line PT67,after G418 screening,and determination of the virus titer,a cell strain with high virus titer(1\^9×10 7)was chosen to transduct ovarian carcinoma cell lines SKOV3,3AO,a transformed cell line permanently expressing HPV16E6E7 was selected respectively,which was identified by RT\|PCR and immonocytochemistry.The drug\|resistant ability to cisplatinum of the transformed cells was tested by means of MTT method. Results\ No change was found in the cisplatinum resistance of the HPV16E6E7 transformed ovarian carcinoma cell lines. Conclusion\ The drug\|resistant changes were not found in HPV16E6E7 transformed ovarian cells.The above study lay a foundation for the establishment of immortal ovarian carcinoma cell lines by means of HPV16E6E7 transduction,especially the drug\|resistant ovarian ones.
出处
《解剖学报》
CAS
CSCD
北大核心
2000年第2期148-151,I015,共5页
Acta Anatomica Sinica
基金
国家自然科学基金!( 3 9870 85 4)
关键词
卵巢肿瘤
耐药性
人乳头瘤病毒
顺铂
Ovarian neoplasms/drug\|resistance
Human Papillomavirus
