Wang Lili, Wang Yanyan, Liu Yajing, Cheng Min, Wu Xu, Wei Haiming
Hefei National Laboratory for Physical Sciences at Microscale and School of Life Sciences, University of Science and Technology of China, Anhui, PR China.
J Exp Clin Cancer Res. 2009 Apr 28;28(1):57. doi: 10.1186/1756-9966-28-57.
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.
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.
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.
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.
免疫细胞化学和逆转录-聚合酶链反应已广泛用于检测乳腺癌患者循环肿瘤细胞,但它们的特异性有限。我们的目的是利用一种便捷且特异的技术检测乳腺癌患者的循环肿瘤细胞。
为确定我们方法的敏感性和特异性,将A431细胞与人外周血白细胞进行系列稀释,并用细胞角蛋白19(CK19)染色。通过流式细胞术检测包括25名健康志愿者以及48名乳腺癌和良性肿瘤患者在内的总共73份血液标本,以量化CK19的表达。
A431细胞的可检测上限为104个人类白细胞中有1个癌细胞。27%的乳腺癌患者检测到CK19,但对照组均无阳性结果。癌细胞数量随疾病分期逐渐增加,其中I期最少(0%),IV期最多(1.29%)。对15例患者术后进行了为期三个月的化疗观察,治疗后其CK19表达水平大多下降。
我们的研究证实,通过流式细胞术检测外周血中的CK19也是监测乳腺癌循环肿瘤细胞的一种特异且可行的方法。