Mechanisms of drug resistance in ovarian cancer

卵巢癌的耐药机制

• 批准号: 8335858
• 负责人: Patrice Morin
• 金额: $ 23万
• 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/8335858
• 关键词: ABCB1 gene; Antineoplastic Agents; Cancer cell line; Candidate Disease Gene; Cell Line; Cells; Cellular biology; Cisplatin; Doxorubicin; Drug resistance; GAGE; Gene Expression; Genes; Immunohistochemistry; Malignant neoplasm of ovary; Modeling; Molecular Biology; PRSS8 gene; Paclitaxel; Patients; Pharmaceutical Preparations; Phenotype; Refractory; Resistance; Reverse Transcriptase Polymerase Chain Reaction; Role; Series; Testing; Tissues; chemotherapy; expression cloning; 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 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.

使用卵巢癌细胞系 OV90 作为起始细胞系，我们开发了一系列对 3 种不同药物（顺铂、阿霉素和紫杉醇）单独耐药的细胞系。获得了每种抗性表型的三个独立克隆，总共得到了 9 个细胞系用于与亲本细胞进行比较（3 个顺铂抗性克隆 OV90C、3 个阿霉素抗性克隆 OV90D 和 3 个紫杉醇抗性克隆 OV90P）。我们使用 Illumina 微阵列分析这些克隆的基因表达，并鉴定出在各种耐​​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种基因。例如，ABCB1 和 GAGE 基因在耐药细胞中持续升高，而 PRSS8 和 MSMB1 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，我们正在使用多种分子和细胞生物学方法研究这些基因在卵巢癌耐药性中的功能作用。