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

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。这项研究是为了收集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‘定向的区域文库，该文库是从培养的人肾小球系膜细胞中通过Mbo限制性内切酶切割而构建的，以避免反映大小或碱基组成的不同的克隆效率。我们从文库中随机选择了1197个克隆，并用自动DNA测序仪对它们进行了测序。序列数据在克隆之间进行比较，并与DNA数据库(NCBI)进行比较。当两个序列显示出90%以上的同源性时，我们认为它们是相同的。在1197个测序克隆中，450个克隆出现2次以上(代表83个不同物种)，747个克隆出现1次或2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，…更多的功能已经为人所知。当我们根据功能对这些已知基因进行分类时，在人类GMC中表达的基因中，很大一部分是分泌蛋白(71个克隆，7个物种)和蛋白质合成(74个克隆，11个物种)。在人类GMC中最丰富的基因是纤维连接蛋白(47个克隆，占总克隆的4.2%)/通过比较不同细胞(如成纤维细胞、主动脉内皮细胞、粒细胞、成骨细胞和HepG2)或组织(如结肠粘膜和肺)的表达谱，可以将活性基因分为管家和具有GMC细胞特异性功能的基因。我们获得了10个克隆，与其他类型的细胞相比，这些克隆在GMC中特异表达。用从几个人体组织中分离的RNA进行Northern印迹分析，在肾脏和培养的人GMC中检测到这些克隆的强烈信号。10个克隆中的一个(GS9422)被证明是一个未知基因的一部分，与丝氨酸蛋白酶抑制物(Serpin)家族同源。总之，本研究提供了一个在人培养的GMC中表达的基因数据库。较少

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