Subbaramaiah Kotha, Norton Larry, Gerald William, Dannenberg Andrew J
Department of Medicine, New York Presbyterian Hospital, Weill Medical College of Cornell University, Memorial Sloan-Kettering Cancer Center, New York, New York 10021, USA.
J Biol Chem. 2002 May 24;277(21):18649-57. doi: 10.1074/jbc.M111415200. Epub 2002 Mar 18.
Markedly increased levels of cyclooxygenase-2 (COX-2) mRNA, protein, and prostaglandin E(2) synthesis were detected in HER-2/neu-transformed human mammary epithelial cells (184B5/HER) compared with its nontransformed partner cell line (184B5). HER-2/neu stimulated COX-2 transcription via the Ras --> Raf --> MAPK pathway. The inductive effects of HER-2/neu were mediated, in part, by enhanced binding of AP-1 (c-Jun, c-Fos, and ATF-2) to the cyclic AMP-response element (-59/-53) of the COX-2 promoter. The potential contribution of the transcription factor PEA3 was also investigated. Elevated levels of PEA3 were detected in 184B5/HER cells. A PEA3 site (-75/-72) was identified juxtaposed to the cyclic AMP-response element. HER-2/neu-mediated activation of the COX-2 promoter was blocked by mutagenizing the PEA3 site or overexpressing antisense to PEA3. To determine whether HER-2/neu status was also a determinant of COX-2 expression in vivo, we compared levels of COX-2 protein in HER-2/neu-positive and -negative human breast cancers. Increased amounts of COX-2 were detected in HER-2/neu-positive tumors. Taken together, these results suggest that closely spaced PEA3 and cyclic AMP-response elements are required for HER-2/neu-mediated induction of COX-2 transcription. The clear relationship between HER-2/neu status and COX-2 expression in human breast tumors suggests that this mechanism is likely to be operative in vivo.
与未转化的配对细胞系(184B5)相比,在HER-2/neu转化的人乳腺上皮细胞(184B5/HER)中检测到环氧合酶-2(COX-2)的信使核糖核酸(mRNA)、蛋白质水平以及前列腺素E2合成显著增加。HER-2/neu通过Ras→Raf→丝裂原活化蛋白激酶(MAPK)途径刺激COX-2转录。HER-2/neu的诱导作用部分是由活化蛋白-1(AP-1,即c-Jun、c-Fos和活化转录因子2(ATF-2))与COX-2启动子的环磷酸腺苷反应元件(-59/-53)增强结合介导的。还研究了转录因子PEA3的潜在作用。在184B5/HER细胞中检测到PEA3水平升高。在环磷酸腺苷反应元件旁发现了一个PEA3位点(-75/-72)。通过诱变PEA3位点或过表达PEA3反义核酸可阻断HER-2/neu介导的COX-2启动子激活。为了确定HER-2/neu状态是否也是体内COX-2表达的决定因素,我们比较了HER-2/neu阳性和阴性人乳腺癌中COX-2蛋白水平。在HER-2/neu阳性肿瘤中检测到COX-2含量增加。综上所述,这些结果表明,紧密相邻的PEA3和环磷酸腺苷反应元件是HER-2/neu介导的COX-2转录诱导所必需的。HER-2/neu状态与人乳腺肿瘤中COX-2表达之间的明确关系表明,这种机制可能在体内起作用。