应用Cell-search System检测肾癌患者循环肿瘤细胞的方法探索

【作者】 周春；

【导师】 李汉忠；

【作者基本信息】 北京协和医学院 ， 临床医学， 2012， 博士

【摘要】 背景与目的循环肿瘤细胞(Circulating Tumor Cell, CTC)已成为现在肿瘤临床邻域的研究热点之一。目前对CTC研究较多且证实临床价值的肿瘤主要是乳腺癌、前列腺癌、结直肠癌。在这些癌症中CTC已证实与肿瘤的无进展生存期(PFS)、总生存期(OS)及治疗监测(MOT)有明确的关系,其潜在的临床价值包括肿瘤诊断和预测临床分期。肾癌是泌尿外科最常见的恶性肿瘤之一。国内外对肾癌CTC检测的研究罕见。本研究拟探索目前国际上检测CTC最准确、最新的方法—细胞搜索系统(Cell-search System, CSS)用以检测肾癌(RCC)患者CTC的可行性,及改良方法。方法选取2012年在北京协和医院泌尿外科住院及复诊的TNM分期Ⅲ期和Ⅳ期的肾癌患者作为研究对象,采取患者外周血之至少7.5ml,15-30℃保存于专用试管内,在96h内送检,CTC标记物选用与检测乳腺癌、前列腺癌、结直肠癌相同的抗细胞角蛋白(CK)8/18/19荧光抗体,用CSS对患者外周血进行CTC定量检测,评价CSS检测转移性肾癌CTC的检出率和可行性,并探讨改进方法。结果1、8例肾细胞癌患者外周血CTC定量检测结果均为阴性。2、1例患者外周血中发现较多CK8/18/19+和CD45+的双阳性细胞。3、用CSS检测Ⅲ期和Ⅳ期肾癌CTC检出率无差异。结论1、用上皮细胞肿瘤标记物抗CK8/18/19抗体标记肾癌CTC, CSS检测Ⅲ期和Ⅳ期肾癌患者血中CTC的方法检出率低,可行性差。2、肾癌CTC进入外周血后肿瘤细胞抗原发生变异,如CK8/18/19低表达甚至不表达,肿瘤细胞表达新抗原。3、不同的上皮肿瘤细胞表达抗原存在差异,抗CK8/18/19抗体不能作为所有上皮来源肿瘤的细胞标记物。4、要修正CSS检测肾癌CTC的方法,需寻找不同于传统上皮肿瘤细胞CK抗体的新标记物。更多还原

【Abstract】 Background and Objective It is now one of the hotspots in the field of clinical oncology to study Circulating Tumor Cell (CTC). Up to now the studies of CTC and its clinical value have focused on breast cancer, prostate cancer and colorectal cancer, in which CTC has been confirmed to concern with progression-free survival(PFS)> overall survival(OS) and monitoring of therapy(MOT). The potential clinical value of CTC includes cancer diagnosis and prediction of clinical stage. Renal cell carcinoma(RCC) is one of the most common malignancies in department of urology. The studies of CTC test in patients with renal cell carcinoma are rare either in home and abroad. In this study, we intend to explore the feasibility of the method testing CTC in patients with renal cell carcinoma by Cell-search System(CSS) which is now the newest and most accurate way internationally, and how to improve it.Method TNM stage Ⅲ and Ⅳ patients with renal cell carcinoma hospitalized in the PUMC urology ward and for further consultation in2012are enrolled in this study. At least7.5ml peripheral blood samples from patients with renal cell carcinoma were collected and saved in special test tube by the temperature of15-30℃. The test has to been done in96hours after the blood sample has been drown. We selected fluorescent anti-CK8/18/19antibodies as CTC makers in renal cell carcinoma patients which are same with the CTC makers with which CTC tests were done by CSS in breast cancer, prostate cancer and colorectal cancer patients. Then the number of CTC was quantitative detected by CSS. The feasibility and detection rate of the method testing CTC in patients with RCC by CSS is to be evaluated, and the improvement of method shall be discussed.Results1.Quantitative test results of CTC in8patients with renal cell carcinoma are negative.2.Quite many CK+CD45+"double positive cells" were found in one patient’s peripheral blood.3.The detection rate of CTC has no difference between TNM stage Ⅲ and Ⅳ patients.Conclusionl.The detection rate of the method which tests CTC in TNM stage Ⅲ and Ⅳ renal cell carcinoma patients using CSS with epithelial tumor cell maker anti-CK8/18/19is zero, and the way is unworkable.2.The antigens vary when CTCs in renal cell carcinoma patients enter peripheral blood circulation, for example, CK8/18/19expression comes to a low level, even Ck8/18/19are not expressed or the tumor cells express new cell makers.3.Different types of cancer cells express different cell makers,so anti-CK8/18/19 antibodies can not be used as cell makers in all epithelial cell cancer.4.New renal carcinoma CTC makers shall be searched which are different from traditional epithelial tumor cell makers anti-CK8/18/19antibodies since the method using traditional tumor cell maker is not practical.更多还原