Ishikawa Tomoyoshi, Miwa Masanao, Uchida Kazuhiko
Department of Biochemistry and Molecular Oncology, University of Tsukuba, Ibaraki, Japan.
Thyroid. 2006 May;16(5):435-42. doi: 10.1089/thy.2006.16.435.
We applied quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) to detect tissue-specific mRNAs in circulating cancer cells for the diagnosis of early-stage cancer. By Northern blotting, the thyroid peroxidase gene (TPO) was strictly expressed in the thyroid. We also used RT-PCR to examine TPO and thyroid stimulating hormone receptor (TSHR) mRNAs in peripheral blood in 33 thyroid papillary carcinoma patients at stages I (23 cases), II (8 cases) and III (3 cases), 49 noncancer patients with benign thyroid diseases, and 20 healthy volunteers. TPO mRNA was detected in 14 of 23 (61%) cases of stage I carcinoma but only 2 of 49 cases with benign thyroid disease. TPO mRNA was not detected in 20 healthy volunteers. By real-time quantitative RT-PCR, the estimated number of thyrocytes in the circulation ranged from 0.24 and 2700 cells per milliliter of whole blood in 7 of 9 patients at stages I and II, and thyrocyte number did not correlate with tumor size or serum thyroglobulin level. Our results might suggest that detection and quantification of tissue-specific mRNAs (e.g., TPO) in peripheral blood could serve as a means to identify potential tumor markers at early stages of cancer.
我们应用定量逆转录聚合酶链反应(RT-PCR)来检测循环癌细胞中的组织特异性mRNA,以用于早期癌症的诊断。通过Northern印迹法,甲状腺过氧化物酶基因(TPO)在甲状腺中严格表达。我们还使用RT-PCR检测了33例I期(23例)、II期(8例)和III期(3例)甲状腺乳头状癌患者、49例患有良性甲状腺疾病的非癌症患者以及20名健康志愿者外周血中的TPO和促甲状腺激素受体(TSHR)mRNA。I期癌的23例患者中有14例(61%)检测到TPO mRNA,但49例良性甲状腺疾病患者中仅2例检测到。20名健康志愿者均未检测到TPO mRNA。通过实时定量RT-PCR,I期和II期的9例患者中有7例每毫升全血中循环甲状腺细胞的估计数量在0.24至2700个细胞之间,且甲状腺细胞数量与肿瘤大小或血清甲状腺球蛋白水平无关。我们的结果可能表明,检测和定量外周血中的组织特异性mRNA(如TPO)可作为在癌症早期识别潜在肿瘤标志物的一种手段。