Cloning of DLM-1, a novel gene that is up-regulated in activated macrophages, using RNA differential display
Cloning of DLM-1, a novel gene that is up-regulated in activated macrophages, using RNA differential display
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DOI：
10.1016/s0378-1119(99)00419-9
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发表时间：
1999-11-15
期刊：
影响因子：
3.5
通讯作者：
Kocher, O
中科院分区：
文献类型：
--
作者：
Fu, YN;Comella, N;Kocher, O
肿瘤与其环境相互作用，对宿主细胞进行重新编程，以诱导血管生成、炎症、免疫和免疫抑制等反应。为了了解这些过程，重要的是识别和分离在宿主组织中诱导表达的新基因，以响应肿瘤。腹水肿瘤为分离这些基因提供了一个有吸引力的模型，因为响应的宿主腹膜衬里组织可以与在腹膜腔内悬浮生长的肿瘤细胞干净地分开。我们在此报道了通过差异显示克隆的新基因DLm-1，它在携带MOT腹水肿瘤的小鼠的腹膜衬里组织中高度上调。小鼠腹膜巨噬细胞在干扰素-γ或脂多糖刺激下也表达大量的DLm-1。干扰素-γ刺激后4h，DLm-1的表达上调明显，但不能被放线菌酮阻断，提示在其转录调控区存在干扰素反应元件。Northern印迹分析在正常小鼠肺、肠上皮、肝脏和胸腺组织中检测到显著水平的DLm-1RNA。原位杂交显示，MOT和HT-29小鼠皮下移植实体瘤中DLm-1在宿主反应性基质细胞中有较强的表达，而在肿瘤细胞中无表达。对全长克隆的序列分析表明，该克隆编码一个约为2.0kD的蛋白质。M-R 44330，具有多个潜在的蛋白激酶C和酪蛋白激酶II磷酸化位点。我们的数据表明，DLm-1在肿瘤的宿主反应等重要过程中发挥作用。(C)1999 Elsevier Science B.V.保留所有权利。
Tumors interact with their environment, reprogramming host cells to induce responses such as angiogenesis, inflammation, immunity and immune suppression. To understand these processes, it is important to identify and isolate new genes whose expression is induced in host tissues in response to tumors. Ascites tumors offer an attractive model for isolating such genes, because responding host peritoneal lining tissues can be cleanly separated from tumor cells growing in suspension within the peritoneal cavity. We here report the cloning by differential display of a novel gene, DLM-1, that is highly up-regulated in the peritoneal lining tissue of mice bearing MOT ascites tumors. Mouse peritoneal macrophages, stimulated by IFN-gamma or LPS, also expressed significant amounts of DLM-1. Up-regulation of DLM-1 became evident by 4 h after stimulation with IFN-gamma and was not blocked by cycloheximide, suggesting the presence of IFN responding elements in its transcription regulation region. DLM-1 RNA was detected at significant levels in normal mouse lung, intestinal epithelium, liver and thymus by Northern blot analysis. In situ hybridization of MOT and HT-29 mouse subcutaneous transplanted solid tumors revealed strong DLM-1 expression in the host reactive stromal cells, but not the tumor cells. Sequence analysis of the full-length cDNA clone revealed that it encodes a protein of approx. M-r 44330 with multiple potential protein kinase C and casein kinase II phosphorylation sites. Our data suggest that DLM-1 plays a role in such important processes as host response in neoplasia. (C) 1999 Elsevier Science B.V. All rights reserved.