乳腺癌循环肿瘤细胞中多巴胺受体（DR）表达水平的临床意义及其作用机制研究

项目名称： 乳腺癌循环肿瘤细胞中多巴胺受体（DR）表达水平的临床意义及其作用机制研究

项目编号： No.81502546

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 张桢珍

作者单位： 中国人民解放军第二军医大学

项目金额： 18万元

中文摘要： 实时监控肿瘤负荷，准确评估肿瘤转移潜能，是降低乳腺癌死亡率的关键。循环肿瘤细胞CTC是一种无创高效的肿瘤负荷监控指标，然而不同表型的CTC转移潜能不尽相同，寻找最具转移性和侵袭性的CTC是其临床研究和应用的关键。循环肿瘤干细胞被认为是少数具有高度生存力和转移性的CTC。多巴胺受体DR是一个潜在的肿瘤干细胞标志物，且不表达于造血干细胞和其他成体干细胞，是一个理想的循环肿瘤干细胞标志物。本项目将CTC阴性富集结合免疫荧光染色和原位荧光杂交，检测乳腺癌CTC的DR表达；用数字PCR技术检测DR各亚型的基因表达，对比患者的病理信息和随访数据，分析乳腺癌CTC中DR表达水平对肿瘤转移的预后指导意义。并对DR阳性和DR阴性的CTC分别进行显微切割和单细胞RNA-Seq，分析DR通路因子及作用机理。针对DR的靶向药物已进入临床验证阶段。本项目的开展将为该靶向药物在乳腺癌中的应用提供用药指导依据。

中文关键词： C21_乳腺肿瘤；循环肿瘤细胞；循环肿瘤干细胞；多巴胺受体；肿瘤转移

英文摘要： Monitoring tumor dynamics and accurate assessment of tumor metastatic potential are critical in reducing breast cancer mortality. Circulating tumor cells (CTC) is a noninvasive tumor biomarker and working well in tumor diagnosis and detection. However, CTCs with different phenotypic characteristics indicate different metastatic potential, and identifying the most aggressive metastatic tumor cells is a key in CTCs clinical research and application. Circulating tumor stem cells are thought to be the few highly viability and metastatic CTCs. Dopamine receptor (DR) is a potential marker of tumor stem cells, and not expressed in normal hematopoietic stem cells and other adult stem cells, which made DR an ideal circulating tumor stem cell marker. We will combine CTC negative enrichment with immunofluorescence staining and fluorescence in situ hybridization to detect the DR expression on CTCs in breast cancer patients; whereas gene expression of DR subtypes in CTC will be detected by digital PCR as well. The DR expression of breast cancer CTC on both protein and gene level will be compared to the patient's pathology information and follow-up data, and the relevance of DR expression in breast cancer CTC and tumor metastasis will be analyzed. DR-positive and DR-negative CTC will be collected using Laser Micro-Desection system, and their transcription data will be detected by RNA-Seq technology to find out the relevant factors and mechanism of DR-related pathways. Targeted drugs for DR molecules have entered clinical validation phase. This project will provide a simple non-invasive method for the application of medication guide for targeted drug in breast cancer.

英文关键词： breast tumor;circulating tumor cell;circulating tumor stem cell;dopamine receptor;tumor metastasis