Endo Y, Sasaki T, Harada F, Noguchi M
Department of Experimental Therapeutics, Kanazawa University.
Jpn J Cancer Res. 1990 Aug;81(8):723-6. doi: 10.1111/j.1349-7006.1990.tb02635.x.
We have established a highly sensitive method for specific detection of metastasized human tumor cells in embryonic chicks using the polymerase chain reaction (PCR). The cells (HT-1080 or KMST-6) were inoculated into the chorioallantoic membrane vein of the chick embryo and DNA of each embryonic organ was extracted. Then, a human beta-globin-related sequence (576 bp) in the DNA from the embryonic liver and lung was specifically amplified and detected by gel electrophoresis and by a specific oligonucleotide probe. The amplified fragments from the liver DNA samples increased gradually from 2 h to 7 days after HT-1080 inoculation. On the other hand, with inoculation of non-tumorigenic human embryonal fibroblast KMST-6 cells, the DNA from the embryonic liver 7 days after inoculation did not show the PCR-amplified product. This detection technique can contribute significantly to the precise detection of microscopic metastasis.
我们建立了一种高度灵敏的方法，利用聚合酶链反应（PCR）在胚胎小鸡中特异性检测转移的人类肿瘤细胞。将细胞（HT-1080或KMST-6）接种到鸡胚的绒毛尿囊膜静脉中，并提取每个胚胎器官的DNA。然后，通过凝胶电泳和特异性寡核苷酸探针，特异性扩增并检测来自胚胎肝脏和肺脏的DNA中的人类β-珠蛋白相关序列（576 bp）。接种HT-1080后2小时至7天，肝脏DNA样本的扩增片段逐渐增加。另一方面，接种非致瘤性人类胚胎成纤维细胞KMST-6后，接种7天后胚胎肝脏的DNA未显示PCR扩增产物。这种检测技术可显著有助于精确检测微小转移。
