Paracrine CCL20 Loop Induces Epithelial-Mesenchymal Transition In Breast Epithelial Cells
Paracrine CCL20 Loop Induces Epithelial-Mesenchymal Transition In Breast Epithelial Cells
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DOI：
10.1002/mc.22360
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发表时间：
2016-07-01
影响因子：
4.6
通讯作者：
Muscella, A.
中科院分区：
文献类型：
--
作者：
Marsigliante, S.;Vetrugno, C.;Muscella, A.
我们以前发现，CCL 20诱导原代培养的健康乳腺细胞增殖和迁移。本研究的目的是探讨CCL 20调节原代培养的健康乳腺上皮细胞的上皮间质转化（EMT）和肿瘤邻近区域的血管生成的假设。关键结果表明，CCL 20（a）下调E-钙粘蛋白和ZO-1;（B）上调N-钙粘蛋白、波形蛋白和Snail表达;（c）增加VEGF的mRNA和分泌;（d）增加血管生成微血管出芽。因此，研究了由CCL 20诱发的信号转导途径。我们发现，NF-κ B p65下调（通过小干扰RNA，siRNA）逆转了CCL 20诱导的Snail，并阻断了波形蛋白和N-钙粘蛋白mRNA的上调。此外，PI 3 K/AKT抑制（通过LY 294002）完全阻断CCL 20诱导的Snail和NF-kB活化。抑制JNK 1/2（通过SP 60125）或PKC-α（通过siRNA）或src（通过PP 1）阻断NF-kB活化和Snail表达，表明这些激酶都在NF-kB/Snail的上游。抑制mTOR（通过雷帕霉素）消除了CCL 20对N-钙粘蛋白和波形蛋白蛋白合成的影响。此外，PKC-δ的siRNA抑制CCL 20诱导的mTOR和S6的磷酸化，增加波形蛋白和N-cadherin的表达，并最终阻断CCL 20诱导的EMT。CCL 20通过使用PKC-α、src、Akt、NF-kB和Snail信号传导增加健康乳腺细胞VEGF的mRNA和分泌。总之，肿瘤细胞通过CCL 20向周围健康细胞发出信号，诱导调节参与EMT的分子的表达，并通过分泌VEGF（血管生成的主要贡献者）直接和间接促进血管生成。(C)2015年威利期刊公司
We previously found that CCL20 induced primarily cultured healthy breast cell proliferation and migration. The objective of this study was to investigate the hypothesis that CCL20 modulated the epithelial-mesenchymal transition (EMT) of primarily cultured healthy breast epithelial cells and the angiogenesis in areas adjacent to the tumor. Key results showed that CCL20 (a) down-regulated E-cadherin and ZO-1; (b) up-regulated N-cadherin, vimentin, and Snail expressions; (c) increased mRNA and secretion of VEGF and (d) increased angiogenic micro vessel sprouting. Thus, the signal transduction pathways evoked by CCL20 were investigated. We showed that NF-kB p65 down-regulation (by small interfering RNA, siRNA) reversed CCL20-induced Snail and blocked the up-regulation of vimentin and N-cadherin mRNAs. Furthermore, PI3K/AKT inhibition (by LY294002) completely blocked CCL20-induced Snail and NF-kB activation. Inhibition of JNK1/2 (by SP60125) or PKC-alpha (by siRNA) or src (by PP1) blocked NF-kB activation and Snail expression suggesting that these kinases are all upstream of NF-kB/Snail. Inhibition of mTOR (by rapamycin) abolished the effects of CCL20 on N-cadherin and vimentin protein synthesis. Furthermore, siRNA of PKC-delta inhibited the phosphorylation of CCL20-induced mTOR and S6, increased vimentin and N-cadherin expressions and, finally, blocked the CCL20 induced-EMT. CCL20 increased mRNA and secretion of VEGF by healthy breast cells by using PKC-alpha, src, Akt, NF-kB, and Snail signalling. In summary, tumor cells signal to the surrounding healthy cells through CCL20 inducing the modulation of the expression of molecules involved in EMT and promoting angiogenesis directly and indirectly through the secretion of VEGF, a major contributor to angiogenesis. (C) 2015 Wiley Periodicals, Inc.