Mechanisms of drug resistance in ovarian cancer

卵巢癌的耐药机制

• 批准号: 7732232
• 负责人: Patrice Morin
• 金额: $ 26.54万
• 依托单位: NATIONAL INSTITUTE ON AGING
• 依托单位国家: 美国
• 资助国家: 美国
• 起止时间: 至
• 项目状态: 未结题
• 来源: https://reporter.nih.gov/project-details/7732232
• 关键词: ABCB1 gene; Cancer cell line; Candidate Disease Gene; Cell Line; Cells; Cisplatin; Cisplatin/Doxorubicin/Paclitaxel; Doxorubicin; Drug resistance; GAGE; Gene Expression; Genes; Immunohistochemistry; Malignant neoplasm of ovary; Modeling; P-Glycoprotein; PRSS8 gene; Paclitaxel; Patients; Pharmaceutical Preparations; Phenotype; Refractory; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Series; Testing; Tissues; chemotherapy; expression cloning; human PRSS8 protein; ovarian neoplasm; tumor

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 planning to validate these genes in patient tissues using RT-PCR and immunohistochemistry.

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。我们还计划使用RT-PCR和免疫组织化学在患者组织中验证这些基因。