Cloning of the genes for human stromelysin and stromelysin 2: differential expression in rheumatoid synovial fibroblasts.
Cloning of the genes for human stromelysin and stromelysin 2: differential expression in rheumatoid synovial fibroblasts.
复制标题
人溶基质素和溶基质素 2 基因的克隆：类风湿滑膜成纤维细胞中的差异表达。
DOI：
10.1021/bi00448a004
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发表时间：
1989
期刊：
影响因子：
2.9
通讯作者：
Brinckerhoff,CE
中科院分区：
文献类型：
--
作者：
Sirum,KL;Brinckerhoff,CE
达特茅斯医学院生物化学和医学系，汉诺威，新罕布什尔州 03756 收稿日期：1989 年 5 月 4 日；修订稿于 1989 年 6 月 30 日收到 摘要：Stromelysin 是参与正常和患病过程中细胞外基质重塑的金属蛋白酶基因家族的成员。类风湿滑膜细胞的原代培养物产生大量金属蛋白酶 mRNA 和蛋白质。我们从类风湿滑膜细胞 cDNA 文库中克隆了人溶基质素 cDNA，并使用该 cDNA 分离了人溶基质素基因和相关基因溶基质素 2。我们对部分基因进行了测序，发现这两个基因都包含在约 14 KB 对 DNA 上。使用含有基质溶解素 2 基因组克隆的外显子片段作为 Northern 印迹分析中的特异性探针，我们证明了基质溶解素和基质溶解素 2 在类风湿滑膜细胞、人包皮成纤维细胞和兔滑膜成纤维细胞中的差异表达。此外，使用与细菌基因氯霉素乙酰转移酶（CAT）连接的溶基质素启动子的嵌合构建体，我们发现肿瘤启动子佛波肉豆蔻酸乙酸酯（PMA）、表皮生长因子（EGF）和白细胞介素10（IL-10）诱导所需的元件包含在307个碱基对片段上，其中包括约270个碱基对（bp）的5'侧翼DNA。人类基质溶解素和基质溶解素 2 基因的克隆、其差异表达的记录以及基质溶解素基因中转录调控区域的鉴定将有助于正常过程和类风湿性关节炎等疾病中金属蛋白酶基因表达的研究。基质溶解素是一种中性金属蛋白酶，能够降解结缔组织的非胶原成分，包括蛋白聚糖、纤连蛋白和层粘连蛋白（Okada 等，1986）。胶原酶是一种相关的金属蛋白酶，具有启动间质胶原分解的独特能力（Harris，1985）。人类胶原酶之间有约 50% 的同源性（Brinckerhoff 等人，1987 年；Whitham 等人，
Departments of Biochemistry and Medicine, Dartmouth Medical School, Hanover, New Hampshire 03756 Received May 4, 1989; Revised Manuscript Received June 30, 1989 abstract: Stromelysin is a member of a gene family of metalloproteinases involved in extracellularmatrix remodeling in normal and diseased processes. Primary cultures of rheumatoid synovialcells produce large amounts of metalloproteinase mRNA and proteins. We cloned a cDNA for human stromelysin from a rheumatoid synovial cell cDNA library, and we used the cDNA to isolate the gene for human stromelysin and a related gene, stromelysin 2. We sequenced parts of the genes and found that both are contained on~ 14 kilobase pairs of DNA. Using an exon-containing fragment of the stromelysin 2 genomic clone as a specific probe in Northern blot analysis, we demonstrate the differential expression of stromelysin and stromelysin 2 in rheumatoid synovialcells, human foreskin fibroblasts, and rabbit synovial fibroblasts. In addition, using chimeric constructs of the stromelysin promoter linked to the bacterial gene chloramphenicol acetyltransferase (CAT), we show that the elements required for the tumor promoter phorbolmyristate acetate (PMA), epidermal growth factor (EGF), and interleukin 10 (IL-10) induction are contained on a 307 base pair fragment which includes~ 270 base pairs (bp) of 5'-flanking DNA. The cloning of the human stromelysin and stromelysin 2 genes, the documentation of their differential expression, and the identification of transcriptional regulatory regions in the stromelysin gene will facilitate the study of me-talloproteinase gene expressionin normal processes andin diseases such as rheumatoid arthritis.Stromelysin is a neutral metalloproteinase that has the ability to degrade non-collagen components of connective tissue including proteoglycans, fibronectin, and laminin (Okada et al., 1986). Collagenase is a related metalloproteinase with the singular ability to initiate breakdown of the interstitial col-lagens (Harris, 1985). The~ 50% homology between the human collagenase (Brinckerhoff et al., 1987; Whitham et al.,