Hayle A J, Darling D L, Taylor A R, Tarin D
Nuffield Department of Pathology, John Radcliffe Hospital, University of Oxford, Headington, UK.
Differentiation. 1993 Oct;54(3):177-89. doi: 10.1111/j.1432-0436.1993.tb01600.x.
From a large series of experiments involving transfer of high molecular weight total genomic DNA from highly metastatic human and mouse tumour cell lines to other mouse tumour cell lines we have derived a few cell lines with greatly augmented metastatic properties. In one of these experiments the transfected cell line (designated AH8 Test) not only colonised the lungs but also formed secondary tumour colonies in several extrapulmonary sites including the skin, skeletal muscles, bone, liver diaphragm, spleen and heart. There were no qualitative and quantitative effects of this magnitude when we used DNA from several non-metastatic or non-tumourigenic sources. Secondary transfection of metastatic capability with DNA obtained from a metastasis formed by one of the primary transfectant lines (AH8 Test) has also been accomplished. Concomitant transfer of human DNA through both transfection cycles in this experiment was confirmed by a variety of methods including Southern blot analysis, in situ hybridisation and polymerase chain reaction (PCR) amplification of DNA using primers recognising human-specific Alu repeat sequences. The findings offer opportunities for the isolation of sequences programming metastatic behaviour and we have cloned and sequenced a fragment of human DNA, which has not been previously characterised, from the transfected cells.
在一系列大量实验中，我们将高分子量的总基因组DNA从高转移性的人类和小鼠肿瘤细胞系转移到其他小鼠肿瘤细胞系，从中获得了一些转移特性大大增强的细胞系。在其中一项实验中，转染后的细胞系（命名为AH8 Test）不仅在肺部形成了殖民地，还在包括皮肤、骨骼肌、骨骼、肝脏、横膈膜、脾脏和心脏在内的多个肺外部位形成了继发性肿瘤殖民地。当我们使用来自几种非转移性或非致瘤性来源的DNA时，并没有出现如此程度的定性和定量影响。我们还完成了用从一个原发性转染细胞系（AH8 Test）形成的转移灶中获得的DNA对转移能力进行的二次转染。在这个实验中，通过包括Southern印迹分析、原位杂交以及使用识别人类特异性Alu重复序列的引物对DNA进行聚合酶链反应（PCR）扩增等多种方法，证实了人类DNA在两个转染周期中的伴随转移。这些发现为分离编码转移行为的序列提供了机会，并且我们已经从转染细胞中克隆并测序了一段此前未被表征的人类DNA片段。
