ANALYSIS OF DRUG RESISTANCE BY FLOW MICROFLUOROCYTOMETRY

流式微荧光细胞术耐药性分析

• 批准号: 3774616
• 负责人: J B TREPEL
• 金额: --
• 依托单位: DIVISION OF CANCER TREATMENT
• 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/3774616
• 关键词: P glycoprotein; drug resistance; flow cytometry; fluorescent dye /probe; folate; gene expression; genetic regulation; human tissue; method development; monoclonal antibody; phenotype; receptor; single cell analysis; thymidylate synthase

We have established assays to examine expression of the drug resistance phenotype using flow cytometric analysis of intact single cells. There are significant advantages to this approach, including the ability to derive rapid, semiquantitative data on tens of thousands of cells and the ability to study the heterogeneity of drug resistance within clinical specimens. The techniques developed include analysis of P-glycoprotein expression using the monoclonal antibody MRK-16, flow cytometric drug cytotoxicity assays based on the esterase activity of surviving cells, expression of thymidylate synthase using monoclonal antibody developed by Drs. Johnston and Allegra, and folate receptor studies using fluoresceinated methotrezate or anti-folate receptor antibody with Drs. Chung and Elwood. In the past year, we have used these techniques in a wide variety of applications in collaborative studies. These applications include examination of the regulation of P-glycoprotein expression in breast cancer cell lines transfected with the gene for protein kinase C alpha or protein kinase C gamma with Drs. Ahmad and Glazer; quantitation of intracellular thymidylate synthase levels in parental and 5-FU- resistant colon and breast cancer cells; examination of the expression of the folate receptor in cells transfected with a folate receptor expression vector with Drs. Chung and Elwood; and analysis of the role of myeloperoxidase in vinca resistance in acute myeloblastic leukemia with Drs. Schlaifer and Myers.

我们已经建立了检测耐药性表达的方法， 使用完整单细胞的流式细胞术分析表型。 那里 是这种方法的显著优点，包括能够 获得了数以万计的细胞的快速半定量数据， 研究临床耐药性异质性的能力 标本 开发的技术包括P-糖蛋白分析 使用单克隆抗体MRK-16、流式细胞术药物 基于存活细胞的酯酶活性的细胞毒性测定， 使用由以下开发的单克隆抗体表达胸苷酸合酶： Drs.约翰斯顿和阿莱格拉，和叶酸受体的研究，使用 荧光甲氨蝶呤或抗叶酸受体抗体与Dr. 钟和埃尔伍德。 在过去的一年里，我们在一个 在协作研究中的广泛应用。 这些应用 包括检查P-糖蛋白表达的调节， 用蛋白激酶C基因转染的乳腺癌细胞系 α或蛋白激酶C γ与Ahmad和Glazer博士;定量 细胞内胸苷酸合成酶水平的亲本和5-FU- 耐药结肠癌和乳腺癌细胞; 在用叶酸受体表达转染的细胞中的叶酸受体 与钟博士和埃尔伍德的矢量;和分析的作用， 髓过氧化物酶在急性髓细胞白血病伴长春新碱耐药中的作用 Drs.施莱费尔和迈尔斯。