Sakaguchi M, Virmani A K, Ashfaq R, Rogers T E, Rathi A, Liu Y, Kodagoda D, Cunningham H T, Gazdar A F
Hamon Center for Therapeutic Oncology Research, University of Texas Southwestern Medical Center, Dallas 75235-8593, USA.
Br J Cancer. 1999 Feb;79(3-4):416-22. doi: 10.1038/sj.bjc.6690065.
We developed a sensitive and specific method for the detection of epithelial cancer cells in effusions with a two-stage molecular-based assay which combined enrichment for cancer cells by immunomagnetic bead selection and reverse transcriptase polymerase chain reaction (RT-PCR) detection of epithelial glycoprotein 2 (EGP-2) RNA. Preliminary experiments indicated that immunobead selection was essential to avoid occasional false-positive RT-PCR results, and this method detected ten breast cancer cells electively added to 10(7) cytologically negative effusion cells. We studied 110 cases of pleural (n = 68) and peritoneal (n = 42) effusions (30 from patients with known carcinoma and 80 from those without known carcinoma), and the results were compared with cytological findings. Of 18 effusions that were cytologically positive or suspicious for malignant cells, 17 (94%) were positive for EGP-2 RNA (the one negative sample was from a patient who recently received combination chemotherapy). Of 92 cytologically negative samples, 11 (12%) were positive for EGP-2, including six patients with a history of previous or current carcinoma. Our method appears to be highly specific and increases the sensitivity of detection of malignant cells; it may be a useful adjunct to routine cytopathological examination.
我们开发了一种灵敏且特异的方法，用于检测积液中的上皮癌细胞，该方法采用两阶段分子检测法，将免疫磁珠筛选富集癌细胞与逆转录聚合酶链反应（RT-PCR）检测上皮糖蛋白2（EGP-2）RNA相结合。初步实验表明，免疫磁珠筛选对于避免偶尔出现的RT-PCR假阳性结果至关重要，且该方法能检测出选择性添加到10⁷个细胞学检查阴性的积液细胞中的10个乳腺癌细胞。我们研究了110例胸腔积液（n = 68）和腹腔积液（n = 42）病例（30例来自已知患有癌症的患者，80例来自无已知癌症的患者），并将结果与细胞学检查结果进行比较。在18例细胞学检查呈阳性或怀疑有恶性细胞的积液中，17例（94%）EGP-2 RNA呈阳性（1例阴性样本来自近期接受联合化疗的患者）。在92例细胞学检查阴性的样本中，11例（12%）EGP-2呈阳性，其中包括6例有既往或当前癌症病史的患者。我们的方法似乎具有高度特异性，并提高了恶性细胞检测的灵敏度；它可能是常规细胞病理学检查的有用辅助手段。
