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Flow cytometric analysis of CK19 expression in the peripheral blood of breast carcinoma patients: relevance for circulating tumor cell detection

机译:乳腺癌患者外周血CK19表达的流式细胞术分析:与循环肿瘤细胞检测的相关性

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Background Immunocytochemistry and RT-PCR have been widely used for the detection of circulating tumor cells in patients with breast cancer but their specificity is limited. Our purpose is to utilize a convenient and specific technology to detect circulating tumor cells in breast cancer patients. Methods To determine the sensitivity and specificity of our method, A431 cells were serially diluted with human peripheral blood leukocytes and stained with CK19. A total of 73 blood specimens including 25 healthy volunteers and 48 patients with breast carcinoma and benign tumor were tested by flow cytometry to quantify the expression of CK19. Results The detectable upper limit of A431 cells was 1 cancer cell among 104 human white blood cells. CK19 was detected in 27% of breast cancer patients but none control gives positive result. The number of cancer cells increased gradually along with the disease stages for it was the least in stage I (0%) and the most in stage IV (1.29%). Fifteen patients were observed during three month chemotherapy after surgery, and most of their CK19 expression levels declined after treatment. Conclusion Our research convinces that the detection of CK19 in peripheral blood by flow cytometry is also a specific and feasible method to monitor circulating tumor cells in breast cancer.
机译:背景技术免疫细胞化学和RT-PCR已被广泛用于检测乳腺癌患者的循环肿瘤细胞,但其特异性有限。我们的目的是利用便捷且特定的技术来检测乳腺癌患者中循环中的肿瘤细胞。方法用人外周血白细胞连续稀释A431细胞,并用CK19染色,以测定本方法的敏感性和特异性。通过流式细胞术检测了73例血液样本,包括25名健康志愿者和48例乳腺癌和良性肿瘤患者,以定量CK19的表达。结果检测到的A431细胞的上限为10 4 人白细胞中的1个癌细胞。在27%的乳腺癌患者中检测到CK19,但没有一个对照组给出阳性结果。癌细胞的数量随着疾病阶段的增加而逐渐增加,因为它在第一阶段最少(0%),在第四阶段最多(1.29%)。术后三个月的化疗期间观察到15名患者,治疗后大多数CK19表达水平下降。结论我们的研究相信,通过流式细胞仪检测外周血中的CK19也是监测乳腺癌中循环肿瘤细胞的一种特定且可行的方法。

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