Zhong X Y, Kaul S, Diel I, Eichler A, Bastert G
Department of Obstetrics and Gynaecology, University Hospital, Heidelberg, Germany.
J Cancer Res Clin Oncol. 1999;125(5):286-91. doi: 10.1007/s004320050275.
The work aimed to evaluate the sensitivity and specificity of the cytokeratin (CK) 19 reverse transcriptase/polymerase chain reaction (RT-PCR) for the detection of occult breast cancer in bone marrow and leukapheresis products.
Peripheral blood and bone marrow samples, obtained from 96 and 8 healthy donors respectively, served as negative controls. A total of 115 bone marrow samples and 29 leukapheresis samples from routine patients with breast cancer were analysed by CK19 RT-PCR. The PCR results were compared with those from routine immunocytology for CK8, 18, 19.
The CK19 RT-PCR technique with primer pairs from Datta et al. (J Clin Oncol 12: 475-482, 1994), using an annealing temperature of 72 degrees C, allowed the detection of one tumour cell in 10(7) mononuclear cells. None of the control samples (96 peripheral blood and 8 bone marrow) that were positive for beta2-microglobulin by RT-PCR showed a signal for CK19. However, expression of CK19 mRNA was observed in 40.87% (70/115) of bone marrow and in 24.13% (7/29) of leukapheresis samples of patients with breast cancer. Standard immunocytology and PCR were combined for the detection of tumour cells. Five of the 65 bone marrow samples were found to be positive by CK19 RT-PCR, but were negative with the immunocytology method.
RT-PCR using CK19-specific primers and optimal experimental conditions is a reliable and specific method for the detection of micrometastatic breast cancer cells.
本研究旨在评估细胞角蛋白（CK）19逆转录酶/聚合酶链反应（RT-PCR）检测骨髓和白细胞分离产物中隐匿性乳腺癌的敏感性和特异性。
分别从96名健康供体和8名健康供体获取外周血和骨髓样本作为阴性对照。对115例乳腺癌常规患者的骨髓样本和29例白细胞分离样本进行CK19 RT-PCR分析。将PCR结果与CK8、18、19的常规免疫细胞学结果进行比较。
采用Datta等人（《临床肿瘤学杂志》12: 475 - 482, 1994）的引物对，退火温度为72℃的CK19 RT-PCR技术能够在10⁷个单核细胞中检测到一个肿瘤细胞。RT-PCR检测β2-微球蛋白呈阳性的对照样本（96例外周血和8例骨髓）均未显示CK19信号。然而，在乳腺癌患者的40.87%（70/115）的骨髓样本和24.13%（7/29）的白细胞分离样本中观察到CK19 mRNA表达。将标准免疫细胞学和PCR结合用于检测肿瘤细胞。65例骨髓样本中有5例经CK19 RT-PCR检测呈阳性，但免疫细胞学方法检测为阴性。
使用CK19特异性引物和最佳实验条件的RT-PCR是检测微转移乳腺癌细胞的可靠且特异的方法。
