Marsigliante S, Vetrugno C, Muscella A
Dipartimento di Scienze e Tecnologie Biologiche e Ambientali (Di.S.Te.B.A.), Laboratorio di Fisiologia Cellulare, Università del Salento, Via Provinciale per Monteroni, Lecce, Italy.
Unità di Neuropatologia, Istituto di Neurologia sperimentale e Divisione di Neuroscienze, Istituto Scientifico IRCCS San Raffaele, Milano, Italy.
Mol Carcinog. 2016 Jul;55(7):1175-86. doi: 10.1002/mc.22360. Epub 2015 Jul 8.
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-α (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-δ 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-α, 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. © 2015 Wiley Periodicals, Inc.
我们之前发现CCL20可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CCL20调节原代培养的健康乳腺上皮细胞上皮-间质转化（EMT）以及肿瘤周边区域血管生成这一假说。主要结果显示，CCL20：（a）下调E-钙黏蛋白和ZO-1；（b）上调N-钙黏蛋白、波形蛋白和Snail的表达；（c）增加VEGF的mRNA水平和分泌量；（d）增加血管生成微血管的出芽。因此，我们对CCL20引发的信号转导通路进行了研究。我们发现，NF-κB p65下调（通过小干扰RNA，即siRNA）可逆转CCL20诱导的Snail表达，并阻断波形蛋白和N-钙黏蛋白mRNA的上调。此外，PI3K/AKT抑制（通过LY294002）可完全阻断CCL20诱导的Snail表达和NF-κB激活。抑制JNK1/2（通过SP60125）或PKC-α（通过siRNA）或src（通过PP1）可阻断NF-κB激活和Snail表达，这表明这些激酶均位于NF-κB/Snail的上游。抑制mTOR（通过雷帕霉素）可消除CCL20对N-钙黏蛋白和波形蛋白蛋白合成所产生的影响。此外，PKC-δ的siRNA可抑制CCL20诱导的mTOR和S6磷酸化，增加波形蛋白和N-钙黏蛋白的表达，并最终阻断CCL20诱导的EMT。CCL20通过PKC-α、src、Akt、NF-κB和Snail信号通路增加健康乳腺细胞VEGF的mRNA水平和分泌量。总之，肿瘤细胞通过CCL20向周围健康细胞发出信号，诱导参与EMT的分子表达发生改变，并通过分泌血管生成的主要促成因子VEGF直接和间接地促进血管生成。© 2015威利期刊公司
