Mechanisms of drug resistance in ovarian cancer

以卵巢癌细胞系OV 90为起始细胞系，我们已经建立了一系列对3种不同药物（顺铂、阿霉素和紫杉醇）具有耐药性的细胞系。获得了每个抗性表型的三个独立克隆，总共得到9个细胞系用于与亲本细胞进行比较（3个顺铂抗性克隆OV 90 C、3个多柔比星抗性克隆OV 90D和3个紫杉醇抗性克隆OV 90 P）。我们使用Illumina微阵列分析这些克隆的基因表达，并确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共有的几个基因。例如，ABCB 1和GAGE基因在耐药细胞中持续升高，而PRSS 8和MSMB 1持续下调。我们目前正在这些细胞系中通过定量RT-PCR验证这些和其他候选基因。我们还使用RT-PCR和免疫组织化学在患者组织中验证这些基因。最后，我们正在研究这些基因在卵巢癌耐药中的功能作用，使用一些分子和细胞生物学方法。

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.