Datta Y H, Adams P T, Drobyski W R, Ethier S P, Terry V H, Roth M S
Department of Medicine, University of Michigan Medical School, Ann Arbor.
J Clin Oncol. 1994 Mar;12(3):475-82. doi: 10.1200/JCO.1994.12.3.475.
Detection of occult carcinoma in patients with breast cancer may aid the establishment of prognosis and development of new therapeutic approaches. To improve on existing methods of detection, we have developed a reverse-transcriptase polymerase chain reaction (RT-PCR) assay for keratin 19 (K19) transcripts to identify mammary carcinoma cells in the peripheral blood and bone marrow of patients with breast cancer.
Peripheral-blood or bone marrow samples obtained from 34 patients with stages I to IV breast cancer and 39 control subjects without breast cancer were screened for K19 mRNA by nested primer PCR.
In reconstitution experiments, K19 RT-PCR reliably detected 10 mammary carcinoma cells in 1 million normal peripheral-blood mononuclear (PBMN) cells. Four of 19 patients with stage IV breast cancer had detectable K19 transcript in peripheral blood. Five of six patients with histologically negative bone marrow biopsies following preablative chemotherapy and before autologous bone marrow transplant (BMT) were positive by this assay. Stem-cell apheresis harvests obtained from one of these patients and three additional patients immediately before BMT were all K19-negative. K19 RT-PCR analysis of CSF from a breast cancer patient with known carcinomatous meningitis was also positive. Thirty-eight of 39 non-breast cancer patients had negative K19 RT-PCR assays. The one exception was a patient with chronic myelogenous leukemia.
RT-PCR of K19 is a sensitive, specific, and rapid method for detection of occult mammary carcinoma cells in the peripheral blood and bone marrow of patients with breast cancer. The presence of residual breast cancer cells in histologically normal bone marrow aspirates but not in stem-cell apheresis harvests is a frequent finding. This assay may be useful in diagnosing metastatic disease, as well as in monitoring the effectiveness of systemic therapy.
检测乳腺癌患者中的隐匿性癌可能有助于判断预后并开发新的治疗方法。为改进现有的检测方法，我们开发了一种用于角蛋白19（K19）转录本的逆转录聚合酶链反应（RT-PCR）检测法，以识别乳腺癌患者外周血和骨髓中的乳腺癌细胞。
通过巢式引物PCR对34例Ⅰ至Ⅳ期乳腺癌患者和39例无乳腺癌的对照受试者的外周血或骨髓样本进行K19 mRNA筛查。
在重组实验中，K19 RT-PCR能可靠地检测出100万个正常外周血单个核（PBMN）细胞中的10个乳腺癌细胞。19例Ⅳ期乳腺癌患者中有4例在外周血中检测到可检测的K19转录本。6例在清髓性化疗后和自体骨髓移植（BMT）前骨髓活检组织学阴性的患者中有5例通过该检测呈阳性。在BMT前立即从其中1例患者和另外3例患者采集的干细胞单采收获物均为K19阴性。对一名已知患有癌性脑膜炎的乳腺癌患者的脑脊液进行K19 RT-PCR分析也呈阳性。39例非乳腺癌患者中有38例K19 RT-PCR检测为阴性。唯一的例外是一名慢性粒细胞白血病患者。
K19的RT-PCR是检测乳腺癌患者外周血和骨髓中隐匿性乳腺癌细胞的一种灵敏、特异且快速的方法。组织学正常的骨髓穿刺物中存在残留乳腺癌细胞，但干细胞单采收获物中不存在，这是常见的发现。该检测法可能有助于诊断转移性疾病，以及监测全身治疗的效果。
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