Cloning and characterization of a growth factor-inducible cyclooxygenase gene from rat intestinal epithelial cells.
Cloning and characterization of a growth factor-inducible cyclooxygenase gene from rat intestinal epithelial cells.
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大鼠肠上皮细胞生长因子诱导环氧合酶基因的克隆和表征。
DOI：
10.1152/ajpgi.1994.266.5.g822
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发表时间：
1994
期刊：
影响因子：
--
通讯作者：
Lanahan,A
中科院分区：
文献类型：
--
作者：
DuBois,RN;Tsujii,M;Bishop,P;Awad,JA;Makita,K;Lanahan,A
生长因子在肠上皮细胞的生长调控和转化中发挥重要作用。利用标准的差异克隆技术，我们从大鼠肠上皮细胞中分离出一个生长因子诱导基因（RS-2），该基因在氨基酸水平上与小鼠丝裂原诱导的环氧化酶（COX-2）具有约95%的同源性。该cDNA与来自转化生长因子(TGF)- α刺激的大鼠肠上皮细胞（RIE-1）的约4.5 kb mRNA杂交，并在成年大鼠肾脏和大脑中组成性表达。核运行实验表明，tgf - α刺激后RS-2 mRNA的增加部分是由于该基因的转录率增加。将RS-2编码区亚克隆到pCMV-2表达载体上，在COS-1细胞中表达RS-2蛋白。从转染RS-2表达载体的COS-1细胞分离的微粒体片段具有环加氧酶活性。除了产生前列腺素外，重组RS-2蛋白还催化了其他三种类二十烷酸产物的形成。综上所述，我们从大鼠肠细胞中克隆了一个生长因子刺激后诱导的丝裂原诱导的环加氧酶基因。
Growth factors have been shown to play a role in intestinal epithelial growth regulation and transformation. Utilizing standard differential cloning techniques, we have isolated a growth factor-inducible gene (RS-2) from rat intestinal epithelial cells that has approximately 95% homology to the mouse mitogen-inducible cyclooxygenase (COX-2) at the amino acid level. This cDNA hybridizes to a approximately 4.5-kb mRNA from transforming growth factor (TGF)-alpha-stimulated rat intestinal epithelial (RIE-1) cells and is constitutively expressed in vivo in adult rat kidney and brain. Nuclear run-on experiments demonstrate that the increase of RS-2 mRNA after TGF-alpha stimulation is in part due to an increased transcription rate of the gene. The coding region for RS-2 was subcloned into a pCMV-2 expression vector, and the RS-2 protein was expressed in COS-1 cells. Microsomal fractions isolated from the COS-1 cells transfected with the RS-2 expression vector contained cyclooxygenase activity. In addition to the production of prostaglandins, the recombinant RS-2 protein also catalyzed the formation of three other eicosanoid products. In summary, we have cloned a mitogen-inducible cyclooxygenase gene from rat intestinal cells that is induced following growth factor stimulation.