Mechanisms of drug resistance in ovarian cancer

以卵巢癌细胞株OV90为起点，建立了一系列对顺铂、阿霉素和紫杉醇三种不同药物分别耐药的细胞系。每种耐药表型各获得3个独立克隆，共获得9个细胞株用于与亲本细胞的比较(3个顺铂抗性克隆OV90C，3个阿霉素抗性克隆OV90D，3个紫杉醇抗性克隆OV90P)。我们使用Illumina微阵列分析了这些克隆的基因表达，并鉴定了在各种耐药表型中差异表达的基因。有趣的是，我们已经确定了几种抗性类型的共同基因。例如，ABCB1和GAGE基因在耐药细胞中持续上调，而PRSS8和MSMB1基因持续下调。我们目前正在通过定量RT-PCR在这些细胞系中验证这些和其他候选基因。我们还在使用RT-PCR和免疫组织化学方法验证患者组织中的这些基因。最后，我们正在使用一些分子和细胞生物学方法研究这些基因在卵巢癌耐药中的功能作用。

Using ovarian cancer cell line OV90 as a starting line, we have developed a series of lines individually resistant to 3 different drugs (cisplatin, doxorubicin and paclitaxel). Three independent clones for each resistant phenotype was obtained, giving us a total of 9 cell lines for comparison to the parental cells (3 cisplatin resistant clones OV90C, 3 doxorubicin resistant clones OV90D, and 3 paclitaxel resistant clones OV90P). We have used Illumina microarrays to analyze gene expression of these clones and have identified genes differentially expressed in the various drug resistance phenotypes. Interestingly, we have identified several genes that are common to several resistance types. For example, ABCB1 and GAGE genes were consistently elevated in resistant cells, while PRSS8 and MSMB1 were consistently downregulated. We are currently validating these and other candidate genes by quantitative RT-PCR in these cell lines. We are also validating these genes in patient tissues using RT-PCR and immunohistochemistry. Finally, we are investigating the functional roles of these genes in ovarian cancer drug resistance using a number of molecular and cell biology approaches.