Sabile A, Louha M, Bonte E, Poussin K, Vona G, Mejean A, Chretien Y, Bougas L, Lacour B, Capron F, Roseto A, Bréchot C, Paterlini-Bréchot P
Institut National de Sante et Recherche Medicale U 370, CHU Necker, Paris, France.
Am J Clin Pathol. 1999 Aug;112(2):171-8. doi: 10.1093/ajcp/112.2.171.
We compared the efficacy of 4 methods for isolating circulating tumor cells: immunocapture with Ber-EP4-coated magnetic beads, density gradient separation, ammonium chloride, and distilled water-mediated erythrocyte lysis. Human blood from healthy volunteers was mixed with serial dilutions of prostate (LNCaP) and liver (HepG2) derived tumor cells. Isolation of circulating tumor cells was followed by reverse transcriptase-polymerase chain reaction with primers specific for prostate-specific antigen and alpha-fetoprotein. Ber-EP4 antigen expression was evaluated by immunohistochemistry in 27 hepatocellular carcinomas and 34 prostate adenocarcinomas. Peripheral blood samples from 12 patients with hepatocellular carcinoma and 10 with prostate adenocarcinoma also were tested. Density gradient separation and Ber-EP4 immunocapture were the most sensitive techniques for isolating circulating tumor cells in in vitro tests. Isolation by density gradient separation was significantly more sensitive than Ber-EP4 immunocapture when applied to peripheral blood samples of patients with cancer, a result consistent with the variable expression of Ber-EP4 antigen that we found by immunohistochemistry in prostate adenocarcinomas and hepatocellular carcinomas.
我们比较了4种分离循环肿瘤细胞的方法的效果：用Ber-EP4包被的磁珠进行免疫捕获、密度梯度分离、氯化铵法以及蒸馏水介导的红细胞裂解。将健康志愿者的血液与前列腺来源（LNCaP）和肝脏来源（HepG2）的肿瘤细胞的系列稀释液混合。分离循环肿瘤细胞后，使用针对前列腺特异性抗原和甲胎蛋白的引物进行逆转录-聚合酶链反应。通过免疫组织化学在27例肝细胞癌和34例前列腺腺癌中评估Ber-EP4抗原表达。还检测了12例肝细胞癌患者和10例前列腺腺癌患者的外周血样本。在体外试验中，密度梯度分离和Ber-EP4免疫捕获是分离循环肿瘤细胞最敏感的技术。当应用于癌症患者的外周血样本时，密度梯度分离法的分离效果明显比Ber-EP4免疫捕获更敏感，这一结果与我们通过免疫组织化学在前列腺腺癌和肝细胞癌中发现的Ber-EP4抗原的可变表达一致。
