QUALITATIVE AND QUANTITATIVE ANALYSIS OF GENES EXPRESSED IN CULTURED HUMAN GLOMERULAR MESANGIAL CELLS.

对培养的人肾小球系膜细胞中表达的基因进行定性和定量分析。

• 批准号: 07457240
• 负责人: MIYATA Toshio
• 金额: $ 4.48万
• 依托单位: NAGOYA UNIVERSITY
• 依托单位国家: 日本
• 项目类别: Grant-in-Aid for Scientific Research (B)
• 财政年份: 1995
• 资助国家: 日本
• 起止时间: 1995 至 1996
• 项目状态: 已结题
• 来源: https://kaken.nii.ac.jp/grant/KAKENHI-PROJECT-07457240/
• 关键词: Glomerular mesangial cells (GMC); gene expression profile; fibronectin; serine protease inhibitor (serpin) family; HNC特異的遺伝子

Glomerular mesangial cells (GMC) are thought to have a pivotal role in the development of glomerular diseases. The present study was undertaken to collect the gene expression profile of GMC,a database that describes which genes are expressed and to what extent. We used a 3'-directed regional cDNA library which was constructed from cultured human GMC by cleaving cDNA with Mbo Irestriction enzyme to avoid variable cloning efficiencies reflecting the size or base composition. We randomly selected 1197 clones from the library and sequenced them with an automatic DNA sequencer. The Sequence data were compared among the clones and also with the DNA databank (NCBI). We regarded two sequences as identical when they showed more than 90 percent homology. Among the 1197 sequenced clones, 450 clones appeared more than twice (representing 83 different species) and 747 clones appeared once or twice (representing 713 different species). Among 450 redundant clones, 344 clones corresponded to 53 genes, … More the functions of which are already known. When we classified these known genes according to their functions, a significant fraction of the genes expressed in human GMC are for secretable proteins (71 clones, 7 species) and for protein synthesis (74 clones, 11 species). The most abundant gene in human GMC is fibronectin (47 clones corresponding to 4.2 percent of total clones)/ A comparison of the expression profiles among different cells such as fibroblast, aortic endothelium, granulocyte, osteoblast, and HepG2, or tissues such as colon mucosa and lung, has allowed active genes to be classified as housekeepers or those with GMC cell-specific functions. We obtained 10 clones which were expressed especially in GMC,as compared to other cell types. By northern blot analysis using RNAs isolated from several human tissues, an intense signal for these clones were detected in kidney as well as cultured human GMC.One of 10 clones (GS9422) was shown to be a part of an unidentified gene with a homology to the serine protease inhibitor (serpin) family. In conclusion, the present study provide a database of genes expressed in human cultured GMC. Less

肾小球系膜细胞（GMC）被认为在肾小球疾病的发展中起着关键作用。本研究旨在收集GMC的基因表达谱，这是一个描述哪些基因表达以及表达程度的数据库。我们使用了一个3 ′-定向区域cDNA文库，该文库是通过用Mbo I限制性酶切割cDNA从培养的人GMC构建的，以避免反映大小或碱基组成的可变克隆效率。我们从文库中随机挑选了1197个克隆，用自动DNA测序仪进行测序。将序列数据在克隆之间进行比较，并与DNA数据库（NCBI）进行比较。当两个序列的同源性超过90%时，我们认为它们是相同的。在1197个测序克隆中，450个克隆出现两次以上（代表83个不同物种），747个克隆出现一次或两次（代表713个不同物种）。在450个冗余克隆中，344个克隆对应53个基因， ...更多信息 其功能是已知的。当我们根据它们的功能对这些已知基因进行分类时，在人GMC中表达的基因的显著部分是用于分泌蛋白（71个克隆，7个物种）和用于蛋白质合成（74个克隆，11个物种）。人GMC中最丰富的基因是纤连蛋白（47个克隆，占总克隆的4.2%）。比较不同细胞（如成纤维细胞、主动脉内皮细胞、粒细胞、成骨细胞和HepG 2）或组织（如结肠粘膜和肺）之间的表达谱，可以将活性基因分类为管家基因或具有GMC细胞特异性功能的基因。我们获得了10个克隆，特别是在GMC中表达，与其他细胞类型相比。通过北方印迹分析，发现这些克隆在肾脏和培养的人GMC中有较强的信号，其中有一个克隆（GS 9422）是一个与丝氨酸蛋白酶抑制剂（serepase inhibitor，serpin）家族同源的未知基因的一部分。总之，本研究提供了一个人类培养GMC中表达基因的数据库。