title: Paracrine CCL20 loop induces epithelial-mesenchymal transition in breast epithelial cells.
abstract: 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.
authors.affiliation: 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.
authors.forename: S
authors.initials: S
authors.lastname: Marsigliante
authors.affiliation: Unità di Neuropatologia, Istituto di Neurologia sperimentale e Divisione di Neuroscienze, Istituto Scientifico IRCCS San Raffaele, Milano, Italy.
authors.forename: C
authors.initials: C
authors.lastname: Vetrugno
authors.affiliation: Dipartimento di Scienze e Tecnologie Biologiche e Ambientali (Di.S.Te.B.A.), Laboratorio di Patologia Molecolare, Università del Salento, Via Provinciale per Monteroni, Lecce, Italy.
authors.forename: A
authors.initials: A
authors.lastname: Muscella
mesh_terms: D000818:Animals; D001943:Breast Neoplasms / Q000235:genetics / Q000378:metabolism*; D002478:Cells, Cultured; D054418:Chemokine CCL20 / Q000235:genetics / Q000378:metabolism*; D004847:Epithelial Cells / Q000166:cytology / Q000378:metabolism* / Q000473:pathology; D058750:Epithelial-Mesenchymal Transition; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D019899:Paracrine Communication; D051381:Rats; D015398:Signal Transduction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
chemical_list: C108355:CCL20 protein, human; D054418:Chemokine CCL20; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A
keywords: CCL20; CCR6; EMT; MAPK; NF-kB; PKC; breast; mTOR; snail