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HPV16E6E7转导的卵巢癌细胞系顺铂耐药性的变化 |
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曹善津 钱和年 冯捷 付天云 叶雪 刘蓓 姚煜 刘广芝
【摘要】 目的 人乳头瘤病毒16型E6E7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HPV16E6E7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HPV16E6E7的重组逆转录病毒载体pLXSN16E6E7,转染逆转录病毒包装细胞PT67,经G418筛选,病毒滴度测定,筛选出1株高滴度(1.9×107/ml)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HPV16E6E7的克隆,SKOV3/E6E7、3AO/E6E7。经RT-PCR、免疫细胞化学检测证实HPV16E6E7的表达,之后采用MTT法检测了HPV16E6E7转导的卵巢癌细胞系对顺铂耐药性的变化。结果 HPV16E6E 7转导的卵巢癌细胞系SKOV3/E6E7、3AO/E6E7对顺铂的耐药性无改变。 结论 HPV16E6E7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。 【关键词】 卵巢肿瘤/耐药性;人乳头瘤病毒 【中图分类号】 Q28 【文献标识码】 A 【文章编号】 0529-1356(2000)02-148
CISPLATIN-RESISTANT CHANGES OF HPV16E6E7 TRANSFORMED OVARIAN CARCINOMA CELL LINES
CAO Shan-jin,QIAN He-nian,FENG Jie,FU Tian-yun,YE Xue,LIU Bei,YAO Yu,LIU Guang-zhi (Gynecologic Oncology,People s Hospital,Beijing Medical University, Beijing 100034, China)
【Abstract】 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(1.9×107)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-[1] [2] 下一页 上一个医学论文: 体外培养的施万细胞在平面和聚乙醇酸纤维网架上的迁移 下一个医学论文: 生长因子对成年大鼠肺泡 型细胞增生的影响
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