应用mRNA差异显示技术筛选卵巢癌耐药相关基因  

The selection of taxol resistance associated genes in ovarian cancer cell line by mRNA DD

作　　者：程国钧[1] 田方[2] 李亚里[1] 李春海[2] 王宏[2] 祝华 杨秀玉[3] 

机构地区：[1]中国人民解放军总医院妇产科,北京100853 [2]中国人民解放军军事医学科学院 [3]北京协和医院妇产科

出　　处：《中华医学杂志》2000年第7期541-543,共3页National Medical Journal of China

摘　　要：目的　筛选卵巢癌泰素耐药相关基因。方法　采用间歇诱导法建立了卵巢癌泰素耐药细胞株Skov3/Taxol 2 5 ;采用mRNADD比较Skov3/Taxol 2 5和敏感细胞Skov3的mRNA表达差异 ,筛选卵巢癌泰素耐药相关基因。结果　Skov3/Taxol 2 5对泰素耐药增加 44倍 ,同时对顺铂也产生了耐药。比较Skov3和Skov3/Taxol 2 5的mRNA ,获得 2 2个差异条带。对其中 5个差异条带进行了DNA序列测定 ,有 3个与已知的基因高度同源 ,分别为tPA、Kiaa0 372和LDLC。 2个为未知新基因片段 ,Genbank登录号分别为AF12 0 32 7和AF12 0 32 8。逆转录 聚合酶链反应 (RT PCR)显示 ,tPA和Kiaa0 372为Skov3/Taxol 2 5特异表达基因 ,而LDLC为假阳性。结论　卵巢癌对泰素耐药后有多个基因表达差异。Objective One of the main reasons of the treatment failure for ovarian cancer is the emergence of drug resistance in which several genes involved. The purpose of the study was to select the genes related to Taxol resistance in ovarian cancer. Methods Resistant ovarian cancer cell line, Skov3/Taxol 25, was established by interval exposure Skov3 to Taxol. mRNA was compared between Skov3 and Skov3/Taxol 25 by mRNA DD. Results Skov3/Taxol 25 showed 44 times increase resisance to Taxol after 12 months of induction. There were 22 new differential bands obtained from Skov3/Taxol 25 as compared with Skov3. 18 bands were special by repeated PCR. In the 5 sub clones, three clones were homologous to the known gene. 105/106 base pair of clone 1 were homologous to tPA (tissue plasminogen activator), 261/268 of clone 3 to Kiaa 0372, and 406/409 of clone 18 to LDLC. The other 2 ESTs were new gene segments, which were banked to Genbank with the number of AF120327 and AF120328. The identification by RT PCR revealed that both tPA and Kiaa0372 were special genes from Skov3/Taxol 25, while LDLC was pseudopositive. Conclusion Several genes were involved in the resistance to Taxol in ovarian cancer. The enhanced expression of tPA indicated the increased ability of metastasis after the acquisition of resistance to taxol.

关 键 词：卵巢肿瘤 抗药性 MRNA差异显示 而药基因 

分 类 号：R737.31[医药卫生—肿瘤]