利用基因芯片技术筛选人卵巢癌耐药差异表达基因

Screening of Diffierential Expression Gene Related with Drugresistance in Human Ovarain Cancer Cell Lines by Gene Chip

[目的]比较卵巢上皮癌铂类耐药和非铂类耐药细胞间的基因表达差异,并进行相关基因的筛选和鉴定。[方法]采用人类全基因组表达谱芯片对卵巢癌铂类耐药细胞SKOV3/CB和亲本细胞SKOV3进行差异基因的筛选和分析;用半定量逆转录聚合酶联反应(RT-PCR)对部分芯片结果进行核对验证。[结果]共分离筛选出差异基因3506个,SKOV3/CB比SKOV3上调的差异基因共有1712个,其中上调10倍以上的共有163个,下调的差异基因共有1794个,其中下调10倍以上的共有70个。SKOV3/CB比SKOV3上调10倍以上的3个差异基因,即ZNF198、ERCC5和BIRC3经半定量RT-PCR验证,表明上述3个基因在耐药细胞株中的表达的确高于敏感细胞株。[结论]卵巢癌的多药耐药是一个多种基因参与的复杂过程,ZNF198、ERCC5和BIRC3参与卵巢癌铂类耐药。

[Objective] To investigate the gene differential expression of platinum-resistance cell lines of ovarian epithelial cancer compared with the parent cell lines,and to screen and identify the associated genes. [Methods] Gene expression profiles were screened and analyzed by using whole genome oligonucleotide microarrays for carboplatin-sensitive ovarian cancer cell lines（SKOV3） and carboplatin-resistant cell lines（SKOV3/CB）,and the candidate genes which related to drug resistance were validated by semi-quantitative reverse transcription polymerase chain reaction（RT-PCR）.[Results] A total of 3506 differential genes were screened out,of which up- and down-regulated genes were 1712 and 1794 respectively in SKOV3/CB. And 163 genes and 70 genes were up-regulated more than 10 times in SKOV3/CB and SKOV3 respectively,which were related to many kinds of genes. And the expression levels of 3 genes,ZNF198,ERCC5 and BIRC3,selected from the more than 10 times in SKOV3/CB were verified by RT-PCR. [Conclusion] Many different kinds of genes are involved in the mechanism of mutidrug resistance.ZNF198,ERCC5 and BIRC3 are related to carboplatin-resistance in ovarian cancer.