Qiao D, Stratagouleas E D, Martinez J D
Arizona Cancer Center, Department of Radiation Oncology, University of Arizona, 1501 North Campbell Avenue, Tucson, AZ 85724, USA.
Carcinogenesis. 2001 Jan;22(1):35-41. doi: 10.1093/carcin/22.1.35.
The bile acid deoxycholic acid (DCA) is a known tumor promoter and it has been suggested that DCA-induced apoptosis plays an important role in colon tumor development. In this study we have characterized the capacity of DCA to stimulate mitogen-activated protein kinase (MAPK) activity and examined the effect that MAPK activity had on DCA-induced apoptosis. Analysis of MAPK activity in DCA-treated HCT116 cells using phosphorylation-specific antibodies and in vitro kinase assays indicated that both the extracellular signal-regulated kinase (ERK) and p38 MAPK (p38), but not the c-Jun N-terminal kinase (JNK), were activated. Using pharmacological inhibitors we determined that only ERK could influence DCA cytotoxicity and that elevated ERK activity could suppress DCA-induced apoptosis. This observation was confirmed genetically. Suppressing ERK activity by overexpressing a dominant negative form of the ERK MAP kinase resulted in increased sensitivity to DCA-induced apoptosis whereas elevated ERK activity artificially produced by overexpression of the wild-type ERK kinase blunted DCA-induced apoptosis. Taken together, our results suggest that DCA can stimulate pro-apoptotic and anti-apoptotic signaling pathways and that sensitivity to DCA-induced apoptosis can be modulated by the ERK MAP kinase.
胆汁酸脱氧胆酸(DCA)是一种已知的肿瘤促进剂,有人认为DCA诱导的细胞凋亡在结肠肿瘤发展中起重要作用。在本研究中,我们已对DCA刺激丝裂原活化蛋白激酶(MAPK)活性的能力进行了表征,并研究了MAPK活性对DCA诱导的细胞凋亡的影响。使用磷酸化特异性抗体和体外激酶测定法分析DCA处理的HCT116细胞中的MAPK活性,结果表明细胞外信号调节激酶(ERK)和p38 MAPK(p38)均被激活,但c-Jun氨基末端激酶(JNK)未被激活。使用药理学抑制剂,我们确定只有ERK能影响DCA的细胞毒性,并且升高的ERK活性可抑制DCA诱导的细胞凋亡。这一观察结果通过遗传学方法得到了证实。通过过表达ERK MAP激酶的显性负性形式来抑制ERK活性,导致对DCA诱导的细胞凋亡的敏感性增加,而通过过表达野生型ERK激酶人工产生的升高的ERK活性则减弱了DCA诱导的细胞凋亡。综上所述,我们的结果表明DCA可以刺激促凋亡和抗凋亡信号通路,并且对DCA诱导的细胞凋亡的敏感性可由ERK MAP激酶调节。