IL-6 induces MUC4 expression through gp130/STAT3 pathway in gastric cancer cell lines
IL-6 induces MUC4 expression through gp130/STAT3 pathway in gastric cancer cell lines
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DOI：
10.1016/j.bbamcr.2008.05.020
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发表时间：
2008-10-01
影响因子：
5.1
通讯作者：
de Bolos, Carme
中科院分区：
文献类型：
--
作者：
Mejias-Luque, Raquel;Peiro, Sandra;de Bolos, Carme
据报道，在幽门螺杆菌感染的受试者中，促炎细胞因子白细胞介素6（IL-6）的胃粘膜水平增加，并且在胃腺癌中，已经检测到肠粘蛋白基因（MUC 2和MUC 4）的上调。为了分析IL-6对肠粘蛋白激活的调节作用，用几种浓度的IL-6处理6种胃癌细胞系不同时间，并评估MUC 2和MUC 4的表达。用定量RT-PCR、Western blot和免疫荧光检测IL-6诱导MUC 4表达。而MUC 2表达不受影响。在受体水平阻断gp 130/STAT 3通路后，MUC 4 mRNA水平降低，并且使用AG 490特异性抑制剂在STAT 3活化水平阻断MUC 4 mRNA水平。MUC 4呈现两个STAT因子的推定结合位点，所述STAT因子可在促炎刺激物如IL-6后调节MUC 4转录。通过EMSA、ChIP和定点突变，我们发现STAT 3与顺式元件在-123/-115处结合，该顺式元件传递IL-6介导的MUC 4转录活性上调。结论：IL-6通过gp 130/STAT 3途径诱导胃癌细胞MUC 4的表达，提示IL-6对胃癌细胞MUC 4基因的激活有直接作用。(c)2008 Elsevier B. V.保留所有权利。
The gastric mucosal levels of the pro-inflammatory cytokine Interleukin 6 (IL-6) have been reported to be increased in Helicobacter pylori-infected subjects and, in gastric adenocarcinomas, the up-regulation of intestinal mucin genes (MUC2 and MUC4) has been detected. To analyse the regulatory effects of IL-6 on the activation of intestinal mucins, six gastric cancer cell lines were treated for different times with several concentrations of IL-6, and the expression of MUC2 and MUC4 was evaluated. IL-6 induced MUC4 expression, detected by quantitative RT-PCR, Western blot and immunofluorescence. and MUC2 expression was not affected. MUC4 mRNA levels decreased after blocking the gp130/STAT3 pathway at the level of the receptor, and at the level of STAT3 activation using the AG490 specific inhibitor. MUC4 presents two putative binding sites for STAT factors that May regulate MUC4 transcription after a pro-inflammatory stimulus as IL-6. By EMSA, ChIP and site-directed mutagenesis we show that STAT3 binds to a cis-element at - 123/-115, that conveys IL-6 mediated up-regulation of MUC4 transcriptional activity. We also demonstrated that p-STAT3 binds to MUC4 promoter and a three-fold increase in p-STAT3 binding was observed after treating GP220 cells with IL-6.In conclusion, IL-6 treatment induced MUC4 expression through the gp130/STAT3 pathway, indicating the direct role of IL-6 on the activation of the intestinal mucin gene MUC4 in gastric cancer cells. (c) 2008 Elsevier B.V. All rights reserved.