[en] Epithelial-to-mesenchymal transition (EMT) programs provide cancer cells with invasive and survival capacities that might favor metastatic dissemination. Whilst signaling cascades triggering EMT have been extensively studied, the impact of EMT on the crosstalk between tumor cells and the tumor microenvironment remains elusive. We aimed to identify EMT-regulated soluble factors that facilitate the recruitment of host cells in the tumor. Our findings indicate that EMT phenotypes relate to the induction of a panel of secreted mediators, namely IL-8, IL-6, sICAM-1, PAI-1 and GM-CSF, and implicate the EMT-transcription factor Snail as a regulator of this process. We further show that EMT-derived soluble factors are pro-angiogenic in vivo (in the mouse ear sponge assay), ex vivo (in the rat aortic ring assay) and in vitro (in a chemotaxis assay). Additionally, conditioned medium from EMT-positive cells stimulates the recruitment of myeloid cells. In a bank of 40 triple-negative breast cancers, tumors presenting features of EMT were significantly more angiogenic and infiltrated by a higher quantity of myeloid cells compared to tumors with little or no EMT. Taken together, our results show that EMT programs trigger the expression of soluble mediators in cancer cells that stimulate angiogenesis and recruit myeloid cells in vivo, which might in turn favor cancer spread.
Xu J, Lamouille S, Derynck R,. TGFβ-induced epithelial to mesenchymal transition. Cell Res 2009; 19: 156-172.
Yang J, Weinberg RA,. Epithelial-mesenchymal transition: at the crossroads of development and tumor metastasis. Dev Cell 2008; 14: 818-829.
Peinado H, Olmeda D, Cano A,. Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer 2007; 7: 415-428.
de Herreros AG, Peiro S, Nassour M, et al., Snail family regulation and epithelial-mesenchymal transitions in breast cancer progression. J Mamm Gland Biol Neoplas 2010; 15: 135-147.
Miyoshi A, Kitajima Y, Sumi K, et al., Snail and SIP1 increase cancer invasion by upregulating MMP family in hepatocellular carcinoma cells. Br J Cancer 2004; 90: 1265-1273.
Bindels S, Mestdagt M, Vandewalle C, et al., Regulation of vimentin by SIP1 in human epithelial breast tumor cells. Oncogene 2006; 25: 4975-4985.
Wang WS, Yang XS, Xia M, et al., Silencing of twist expression by RNA interference suppresses epithelial-mesenchymal transition, invasion, and metastasis of ovarian cancer. Asian Pac J Cancer Prev 2012; 13: 4435-4439.
De Craene B, Berx G,. Regulatory networks defining EMT during cancer initiation and progression. Nat Rev Cancer 2013; 13: 97-110.
Chiang C, Ayyanathan K,. Snail/Gfi-1 (SNAG) family zinc finger proteins in transcription regulation, chromatin dynamics, cell signaling, development, and disease. Cytokine Growth Factor Rev 2013; 24: 123-131.
Blanco MJ, Moreno-Bueno G, Sarrio D, et al., Correlation of Snail expression with histological grade and lymph node status in breast carcinomas. Oncogene 2002; 21: 3241-3246.
Blechschmidt K, Sassen S, Schmalfeldt B, et al., The E-cadherin repressor Snail is associated with lower overall survival of ovarian cancer patients. Br J Cancer 2008; 98: 489-495.
Du F, Nakamura Y, Tan TL, et al., Expression of snail in epidermal keratinocytes promotes cutaneous inflammation and hyperplasia conducive to tumor formation. Cancer Res 2010; 70: 10080-10089.
Kudo-Saito C, Shirako H, Takeuchi T, et al., Cancer metastasis is accelerated through immunosuppression during Snail-induced EMT of cancer cells. Cancer Cell 2009; 15: 195-206.
Peinado H, Marin F, Cubillo E, et al., Snail and E47 repressors of E-cadherin induce distinct invasive and angiogenic properties in vivo. J Cell Sci 2004; 117: 2827-2839.
Hwang WL, Yang MH, Tsai ML, et al., SNAIL regulates interleukin-8 expression, stem cell-like activity, and tumorigenicity of human colorectal carcinoma cells. Gastroenterology 2011; 141: 279-291, e271-275.
Bonnomet A, Brysse A, Tachsidis A, et al., Epithelial-to-mesenchymal transitions and circulating tumor cells. J Mamm Gland Biol Neoplas 2010; 15: 261-273.
Bonnomet A, Syne L, Brysse A, et al., A dynamic in vivo model of epithelial-to-mesenchymal transitions in circulating tumor cells and metastases of breast cancer. Oncogene 2012; 31: 3741-3753.
Christiansen JJ, Rajasekaran AK,. Reassessing epithelial to mesenchymal transition as a prerequisite for carcinoma invasion and metastasis. Cancer Res 2006; 66: 8319-8326.
Tsai JH, Donaher JL, Murphy DA, et al., Spatiotemporal regulation of epithelial-mesenchymal transition is essential for squamous cell carcinoma metastasis. Cancer Cell 2012; 22: 725-736.
Chaffer CL, Brennan JP, Slavin JL, et al., Mesenchymal-to-epithelial transition facilitates bladder cancer metastasis: role of fibroblast growth factor receptor-2. Cancer Res 2006; 66: 11271-11278.
Aktas B, Tewes M, Fehm T, et al., Stem cell and epithelial-mesenchymal transition markers are frequently overexpressed in circulating tumor cells of metastatic breast cancer patients. Breast Cancer Res 2009; 11: R46.
Yu M, Bardia A, Wittner BS, et al., Circulating breast tumor cells exhibit dynamic changes in epithelial and mesenchymal composition. Science 2013; 339: 580-584.
Lesniak D, Sabri S, Xu Y, et al., Spontaneous epithelial-mesenchymal transition and resistance to HER-2-targeted therapies in HER-2-positive luminal breast cancer. PLoS One 2013; 8: e71987.
Sarrio D, Rodriguez-Pinilla SM, Hardisson D, et al., Epithelial-mesenchymal transition in breast cancer relates to the basal-like phenotype. Cancer Res 2008; 68: 989-997.
Taube JH, Herschkowitz JI, Komurov K, et al., Core epithelial-to-mesenchymal transition interactome gene-expression signature is associated with claudin-low and metaplastic breast cancer subtypes. Proc Natl Acad Sci USA 2010; 107: 15449-15454.
Sethi S, Sarkar FH, Ahmed Q, et al., Molecular markers of epithelial-to-mesenchymal transition are associated with tumor aggressiveness in breast carcinoma. Transl Oncol 2011; 4: 222-226.
Herold CI, Anders CK,. New targets for triple-negative breast cancer. Oncology 2013; 27: 846-854.
Singh S, Sadanandam A, Singh RK,. Chemokines in tumor angiogenesis and metastasis. Cancer Metast Rev 2007; 26: 453-467.
Waugh DJ, Wilson C,. The interleukin-8 pathway in cancer. Clin Cancer Res 2008; 14: 6735-6741.
Gerber PA, Hippe A, Buhren BA, et al., Chemokines in tumor-associated angiogenesis. Biol Chem 2009; 390: 1213-1223.
Wilson J, Balkwill F,. The role of cytokines in the epithelial cancer microenvironment. Semin Cancer Biol 2002; 12: 113-120.
Chavey C, Bibeau F, Gourgou-Bourgade S, et al., Oestrogen receptor negative breast cancers exhibit high cytokine content. Breast Cancer Res 2007; 9: R15.
Soria G, Ben-Baruch A,. The inflammatory chemokines CCL2 and CCL5 in breast cancer. Cancer Lett 2008; 267: 271-285.
Lenoir B, Wagner DR, Blacher S, et al., Effects of adenosine on lymphangiogenesis. PLoS One 2014; 9: e92715.
Berndt S, Blacher S, Perrier d'Hauterive S, et al., Chorionic gonadotropin stimulation of angiogenesis and pericyte recruitment. J Clin Endocrinol Metab 2009; 94: 4567-4574.
Zhou BP, Deng J, Xia W, et al., Dual regulation of Snail by GSK-3β-mediated phosphorylation in control of epithelial-mesenchymal transition. Nat Cell Biol 2004; 6: 931-940.
Heidemann J, Ogawa H, Dwinell MB, et al., Angiogenic effects of interleukin 8 (CXCL8) in human intestinal microvascular endothelial cells are mediated by CXCR2. J Biol Chem 2003; 278: 8508-8515.
Koch AE, Polverini PJ, Kunkel SL, et al., Interleukin-8 as a macrophage-derived mediator of angiogenesis. Science 1992; 258: 1798-1801.
Li A, Dubey S, Varney ML, et al., IL-8 directly enhanced endothelial cell survival, proliferation, and matrix metalloproteinases production and regulated angiogenesis. J Immunol 2003; 170: 3369-3376.
Gho YS, Kleinman HK, Sosne G,. Angiogenic activity of human soluble intercellular adhesion molecule-1. Cancer Res 1999; 59: 5128-5132.
Wilkins-Port CE, Higgins PJ,. Regulation of extracellular matrix remodeling following transforming growth factor-β1/epidermal growth factor-stimulated epithelial-mesenchymal transition in human premalignant keratinocytes. Cells Tissues Organs 2007; 185: 116-122.
Brysse A, Mestdagt M, Polette M, et al., Regulation of CXCL8/IL-8 expression by zonula occludens-1 in human breast cancer cells. Mol Cancer Res 2012; 10: 121-132.
Bates RC, DeLeo MJ, 3rd, Mercurio AM,. The epithelial-mesenchymal transition of colon carcinoma involves expression of IL-8 and CXCR-1-mediated chemotaxis. Exp Cell Res 2004; 299: 315-324.
Yu J, Ren X, Chen Y, et al., Dysfunctional activation of neurotensin/IL-8 pathway in hepatocellular carcinoma is associated with increased inflammatory response in microenvironment, more epithelial-mesenchymal transition in cancer and worse prognosis in patients. PLoS One 2013; 8: e56069.
Freytag J, Wilkins-Port CE, Higgins CE, et al., PAI-1 mediates the TGF-β1 + EGF-induced 'scatter' response in transformed human keratinocytes. J Invest Dermatol 2010; 130: 2179-2190.
Su S, Liu Q, Chen J, et al., A positive feedback loop between mesenchymal-like cancer cells and macrophages is essential to breast cancer metastasis. Cancer Cell 2014; 25: 605-620.
Morishita Y, Watanabe M, Nakazawa E, et al., The interaction of LFA-1 on mononuclear cells and ICAM-1 on tubular epithelial cells accelerates TGF-β1-induced renal epithelial-mesenchymal transition. PLoS One 2011; 6: e23267.
Li Q, Liu BC, Lv LL, et al., Monocytes induce proximal tubular epithelial-mesenchymal transition through NF-κB dependent upregulation of ICAM-1. J Cell Biochem 2011; 112: 1585-1592.
Fernando RI, Castillo MD, Litzinger M, et al., IL-8 signaling plays a critical role in the epithelial-mesenchymal transition of human carcinoma cells. Cancer Res 2011; 71: 5296-5306.
Sullivan NJ, Sasser AK, Axel AE, et al., Interleukin-6 induces an epithelial-mesenchymal transition phenotype in human breast cancer cells. Oncogene 2009; 28: 2940-2947.
Senoo T, Hattori N, Tanimoto T, et al., Suppression of plasminogen activator inhibitor-1 by RNA interference attenuates pulmonary fibrosis. Thorax 2010; 65: 334-340.
Lim S, Becker A, Zimmer A, et al., SNAI1-mediated epithelial-mesenchymal transition confers chemoresistance and cellular plasticity by regulating genes involved in cell death and stem cell maintenance. PLoS One 2013; 8: e66558.
Lyons JG, Patel V, Roue NC, et al., Snail up-regulates proinflammatory mediators and inhibits differentiation in oral keratinocytes. Cancer Res 2008; 68: 4525-4530.
Fabre-Guillevin E, Malo M, Cartier-Michaud A, et al., PAI-1 and functional blockade of SNAI1 in breast cancer cell migration. Breast Cancer Res 2008; 10: R100.
Li S, Kendall SE, Raices R, et al., TWIST1 associates with NF-κB subunit RELA via carboxyl-terminal WR domain to promote cell autonomous invasion through IL8 production. BMC Biol 2012; 10: 73.
Fantozzi A, Gruber DC, Pisarsky L, et al., VEGF-mediated angiogenesis links EMT-induced cancer stemness to tumor initiation. Cancer Res 2014; 74: 1566-1575.
Abdulkhalek S, Geen O, Brodhagen L, et al., Transcriptional factor snail controls tumor neovascularization, growth and metastasis in mouse model of human ovarian carcinoma. Clin Translat Med 2014; 3: 28.
Olmeda D, Jorda M, Peinado H, et al., Snail silencing effectively suppresses tumour growth and invasiveness. Oncogene 2007; 26: 1862-1874.
Nilsson MB, Langley RR, Fidler IJ,. Interleukin-6, secreted by human ovarian carcinoma cells, is a potent proangiogenic cytokine. Cancer Res 2005; 65: 10794-10800.
Hernandez-Rodriguez J, Segarra M, Vilardell C, et al., Elevated production of interleukin-6 is associated with a lower incidence of disease-related ischemic events in patients with giant-cell arteritis: angiogenic activity of interleukin-6 as a potential protective mechanism. Circulation 2003; 107: 2428-2434.
Nagasaki T, Hara M, Nakanishi H, et al., Interleukin-6 released by colon cancer-associated fibroblasts is critical for tumour angiogenesis: anti-interleukin-6 receptor antibody suppressed angiogenesis and inhibited tumour-stroma interaction. Br J Cancer 2014; 110: 469-478.
Bajou K, Maillard C, Jost M, et al., Host-derived plasminogen activator inhibitor-1 (PAI-1) concentration is critical for in vivo tumoral angiogenesis and growth. Oncogene 2004; 23: 6986-6990.
Bajou K, Noel A, Gerard RD, et al., Absence of host plasminogen activator inhibitor 1 prevents cancer invasion and vascularization. Nat Med 1998; 4: 923-928.
Bajou K, Herkenne S, Thijssen VL,. PAI-1 mediates the antiangiogenic and profibrinolytic effects of 16 K prolactin. 2014; 20: 741-747.
Asfaha S, Dubeykovskiy AN, Tomita H, et al., Mice that express human interleukin-8 have increased mobilization of immature myeloid cells, which exacerbates inflammation and accelerates colon carcinogenesis. Gastroenterology 2013; 144: 155-166.
Cole S, Montero A, Garret-Mayer E, et al., Elevated circulating myeloid derived suppressor cells (MDSC) are associated with inferior overall survival (OS) and correlate with circulating tumor cells (CTC) in patients with metastatic breast cancer. Cancer Res 2010; 69: 4135-4135.
Diaz-Montero CM, Salem ML, Nishimura MI, et al., Increased circulating myeloid-derived suppressor cells correlate with clinical cancer stage, metastatic tumor burden, and doxorubicin-cyclophosphamide chemotherapy. Cancer Immunol Immunother 2009; 58: 49-59.
Toh B, Wang X, Keeble J, et al., Mesenchymal transition and dissemination of cancer cells is driven by myeloid-derived suppressor cells infiltrating the primary tumor. PLoS Biol 2011; 9: e1001162.
Yang L, DeBusk LM, Fukuda K, et al., Expansion of myeloid immune suppressor Gr + CD11b + cells in tumor-bearing host directly promotes tumor angiogenesis. Cancer Cell 2004; 6: 409-421.
Huang Y, Yuan J, Righi E, et al., Vascular normalizing doses of antiangiogenic treatment reprogram the immunosuppressive tumor microenvironment and enhance immunotherapy. Proc Natl Acad Sci USA 2012; 109: 17561-17566.
Morales JK, Kmieciak M, Knutson KL, et al., GM-CSF is one of the main breast tumor-derived soluble factors involved in the differentiation of CD11b-Gr1-bone marrow progenitor cells into myeloid-derived suppressor cells. Breast Cancer Res Treatment 2010; 123: 39-49.
Serafini P, Carbley R, Noonan KA, et al., High-dose granulocyte-macrophage colony-stimulating factor-producing vaccines impair the immune response through the recruitment of myeloid suppressor cells. Cancer Res 2004; 64: 6337-6343.