HUMAN CYCLOOXYGENASE-2 CDNA
HUMAN CYCLOOXYGENASE-2 CDNA
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DOI：
10.1073/pnas.89.16.7384
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发表时间：
1992-08-15
影响因子：
11.1
通讯作者：
NEILSON, K
中科院分区：
文献类型：
--
作者：
HLA, T;NEILSON, K
环氧合酶（Cox），也被称为前列腺素（PG） H合成酶（EC 1.14.99.1），催化炎症性PG形成的限速步骤。PG生物合成的一个主要调控步骤是在Cox水平上：生长因子、细胞因子和肿瘤启动子诱导Cox活性。我们从人脐静脉内皮细胞（HUVEC）中克隆了第二种形式的Cox基因（Cox-2）。该cDNA编码604个氨基酸的多肽，与先前分离的人类Cox-1多肽相同61%。人(h)Cox-2转录物在兔网织细胞裂解物中的体外翻译导致了一个70 kda的蛋白的合成，该蛋白被抗血清免疫沉淀到羊Cox。hCox-2开放阅读框在Cos-7猴肾细胞中的表达导致环加氧酶活性的提高。hCox-2 cDNA在HUVEC中杂交到一个4.5千碱基的mRNA物种，而hCox-1 cDNA杂交到3千碱基和5.3千碱基的mRNA物种。Cox-1和Cox-2 mrna均在HUVEC、血管平滑肌细胞、单核细胞和成纤维细胞中表达。在人内皮细胞和单核细胞中，12-肉豆蔻酸13-醋酸酯和脂多糖优先诱导Cox-2 mRNA表达。总之，这些数据表明，Cox酶是由至少两个基因编码的，这些基因在各种细胞类型中表达和差异调节。hCox-2转录物在间充质源性炎症细胞中的高水平诱导表明在炎症条件中起作用。
Cyclooxygenase (Cox), also known as prostaglandin (PG) H synthase (EC 1.14.99.1), catalyzes the rate-limiting step in the formation of inflammatory PGs. A major regulatory step in PG biosynthesis is at the level of Cox: growth factors, cytokines, and tumor promoters induce Cox activity. We have cloned the second form of the Cox gene (Cox-2) from human umbilical vein endothelial cells (HUVEC). The cDNA encodes a polypeptide of 604 amino acids that is 61 % identical to the previously isolated human Cox-1 polypeptide. In vitro translation of the human (h)Cox-2 transcript in rabbit reticulocyte lysates resulted in the synthesis of a 70-kDa protein that is immunoprecipitated by antiserum to ovine Cox. Expression of the hCox-2 open reading frame in Cos-7 monkey kidney cells results in the elaboration of cyclooxygenase activity. hCox-2 cDNA hybridizes to a 4.5-kilobase mRNA species in HUVEC, whereas the hCox-1 cDNA hybridizes to 3- and 5.3-kilobase species. Both Cox-1 and Cox-2 mRNAs are expressed in HUVEC, vascular smooth muscle cells, monocytes, and fibroblasts. Cox-2 mRNA was preferentially induced by phorbol 12-myristate 13-acetate and lipopolysaccharide in human endothelial cells and monocytes. Together, these data demonstrate that the Cox enzyme is encoded by at least two genes that are expressed and differentially regulated in a variety of cell types. High-level induction of the hCox-2 transcript in mesenchymal-derived inflammatory cells suggests a role in inflammatory conditions.