Mesenchymal stem cells enhance ovarian cancer cell infiltration through IL6 secretion in an amniochorionic membrane based 3D model

Journal of Translational Medicine

Arash Rafii³  Massoud Mirshahi⁴  Eliane Mery¹  Hamda Althawadi²  Mohamed Warfa²  Christophe M Raynaud²  Halema Al Farsi²  Raphael Lis⁵  Cyril Touboul⁴ 

[1] Department of Pathology, Institut Claudius Regaud, Toulouse, France;Stem cell and microenvironment laboratory, Weill Cornell Medical College in Qatar, Education City, Qatar Foundation, Doha, Qatar;Department of Genetic Medicine and Obstetrics and Gynecology, Weill Cornell Medical College, Stem cell and microenvironment laboratory, Weill Cornell Medical College in Qatar, Qatar-Foundation, 24144, Doha, Qatar;UMRS 872 INSERM, Université Pierre et Marie Curie, Equipe 18, Centre de Recherche des Cordeliers, 15 rue de l’Ecole de Medecine, Paris Cedex 06, 75270, France;Department Genetic Medicine, Weill Cornell Medical College, New York, USA

关键词: Mesenchymal stem cell;    3d model;    Tumor infiltration;    IL6;    Ovarian cancer;   

Others  :  1205884
DOI  :  10.1186/1479-5876-11-28

 received in 2012-10-23, accepted in 2013-01-09,  发布年份 2013

【 摘 要 】

Background

The early peritoneal invasion of epithelial ovarian cancer (EOC) by tumoral aggregates presents in ascites is a major concern. The role of the microenvironment seems to be important in this process but the lack of adequate models to study cellular interactions between cancer cells and stromal cells does not allow to uncover the molecular pathways involved. Our goal was to study the interactions between ovarian cancer cells (OCC) and mesenchymal stem cells (MSC) using a 3D model.

Methods

We used millimetric pieces of amniochorionic membrane - referred to as amniotic membrane scaffold (AMS) - to create 3D peritoneal nodules mimicking EOC early invasion. We were able to measure the distribution and the depth of infiltration using confocal microsopy. We extracted MSC from the amniochorionic membrane using the markers CD34-, CD45-, CD73+, CD90+, CD105+ and CD29+ at the Fluorescence Activated Cell Sorting (FACS) analysis. We used transwell and wound healing tests to test OCC migration and invasion in vitro.

Results

Here we show that OCC tumors were located in regions rich in MSC (70%). The tumors infiltrated deeper within AMS in regions rich in MSC (p<0.001). In vitro tests revealed that higher IL6 secretion in a context of MSC-OCC co-culture could enhance migration and invasion of OCC. After IL6 receptor antagonism, OCC infiltration was significantly decreased, mostly in regions rich in MSCs, indicating that recruitment and tridimensional invasion of OCC was dependent of IL6 secretion.

Conclusions

The use of tridimensional models using AMS could be a useful tool to decipher early molecular events in ovarian cancer metastasis. Cytokine inhibitors interrupting the cross-talk between OCCs and MSCs such as IL6 should be investigated as a new therapeutic approach in ovarian cancer.

【 授权许可】

   
2013 Touboul et al.; licensee BioMed Central Ltd.

【 图 表 】

Figure 1.

Figure 2.

Figure 3.

Figure 4.

Figure 5.

【 参考文献 】

• [1]Cannistra SA: Cancer of the ovary. N Engl J Med 2004, 351:2519-2529.
• [2]Tretarre B, Remontet L, Menegoz F, Mace-Lesec'h J, Grosclaude P, Buemi A, Guizard AV, Velten M, Arveux P, Peng J, et al.: Ovarian cancer: incidence and mortality in France. J Gynecol Obstet Biol Reprod (Paris) 2005, 34:154-161.
• [3]Pfisterer J, Ledermann JA: Management of Platinum-Sensitive Recurrent Ovarian Cancer. Semin Oncol 2006, 33:12-16.
• [4]Ceelen WP, Bracke ME: Peritoneal minimal residual disease in colorectal cancer: mechanisms, prevention, and treatment. Lancet Oncol 2009, 10:72-79.
• [5]Bourguignon LY, Gilad E, Peyrollier K: Heregulin-mediated ErbB2-ERK signaling activates hyaluronan synthases leading to CD44-dependent ovarian tumor cell growth and migration. J Biol Chem 2007, 282:19426-19441.
• [6]Lis R, Touboul C, Raynaud CM, Malek JA, Suhre K, Mirshahi M, Rafii A: Mesenchymal cell interaction with ovarian cancer cells triggers pro-metastatic properties. PLoS One 2012, 7:e38340.
• [7]Lis R, Touboul C, Mirshahi P, Ali F, Mathew S, Nolan DJ, Maleki M, Abdalla SA, Raynaud CM, Querleu D, et al.: Tumor associated mesenchymal stem cells protects ovarian cancer cells from hyperthermia through CXCL12. Int J Cancer 2011, 128:715-725.
• [8]Rafii A, Mirshahi P, Poupot M, Faussat AM, Simon A, Ducros E, Mery E, Couderc B, Lis R, Capdet J, et al.: Oncologic trogocytosis of an original stromal cells induces chemoresistance of ovarian tumours. PLoS One 2008, 3:e3894.
• [9]Yamada KM, Cukierman E: Modeling tissue morphogenesis and cancer in 3D. Cell 2007, 130:601-610.
• [10]Freedman RS, Deavers M, Liu J, Wang E: Peritoneal inflammation - A microenvironment for Epithelial Ovarian Cancer (EOC). J Transl Med 2004, 2:23. BioMed Central Full Text
• [11]Ilancheran S, Moodley Y, Manuelpillai U: Human fetal membranes: a source of stem cells for tissue regeneration and repair? Placenta 2009, 30:2-10.
• [12]van der Linden PJ, de Goeij AF, Dunselman GA, Erkens HW, Evers JL: Endometrial cell adhesion in an in vitro model using intact amniotic membranes. Fertil Steril 1996, 65:76-80.
• [13]Groothuis PG, Koks CA, de Goeij AF, Dunselman GA, Arends JW, Evers JL: Adhesion of human endometrium to the epithelial lining and extracellular matrix of amnion in vitro: an electron microscopic study. Hum Reprod 1998, 13:2275-2281.
• [14]Raynaud CM, Maleki M, Lis R, Ahmed B, Al-Azwani I, Malek J, Safadi FF, Rafii A: Comprehensive characterization of mesenchymal stem cells from human placenta and fetal membrane and their response to osteoactivin stimulation. Stem Cells Int 2012, 2012:658356.
• [15]Charles KA, Kulbe H, Soper R, Escorcio-Correia M, Lawrence T, Schultheis A, Chakravarty P, Thompson RG, Kollias G, Smyth JF, et al.: The tumor-promoting actions of TNF-alpha involve TNFR1 and IL-17 in ovarian cancer in mice and humans. J Clin Invest 2009, 119:3011-3023.
• [16]Kenny HA, Kaur S, Coussens LM, Lengyel E: The initial steps of ovarian cancer cell metastasis are mediated by MMP-2 cleavage of vitronectin and fibronectin. J Clin Invest 2008, 118:1367-1379.
• [17]Kenny H, Nieman K, Mitra A, Lengyel E: The First Line of Intra-abdominal Metastatic Attack: Breaching the Mesothelial Cell Layer. Cancer Discovery 2011, 1:100-102.
• [18]Iwanicki M, Davidowitz R, Ng M, Besser A, Muranem T, Merritt M, Danuser G, Ince T, Brugge J: Ovarian Cancer Spheroids Use Myosin-Generated Force to Clear the mesothelium. Cancer Discovery 2011, 1:144-147.
• [19]Coffelt SB, Marini FC, Watson K, Zwezdaryk KJ, Dembinski JL, LaMarca HL, Tomchuck SL, zu Bentrup KH, Danka ES, Henkle SL, Scandurro AB: The pro-inflammatory peptide LL-37 promotes ovarian tumor progression through recruitment of multipotent mesenchymal stromal cells. Proc Natl Acad Sci 2009, 106:3806-3811.
• [20]Jeon ES, Moon HJ, Lee MJ, Song HY, Kim YM, Cho M, Suh D-S, Yoon M-S, Chang CL, Jung JS, Kim JH: Cancer-Derived Lysophosphatidic Acid Stimulates Differentiation of Human Mesenchymal Stem Cells to Myofibroblast-Like Cells. Stem Cells 2008, 26:789-797.
• [21]Seo JH, Jeong KJ, Oh WJ, Sul HJ, Sohn JS, Kim YK, Cho do Y, Kang JK, Park CG, Lee HY: Lysophosphatidic acid induces STAT3 phosphorylation and ovarian cancer cell motility: their inhibition by curcumin. Cancer Lett 2010, 288:50-56.
• [22]Kidd S, Spaeth E, Watson K, Burks J, Lu H, Klopp A, Andreeff M, Marini FC: Origins of the tumor microenvironment: quantitative assessment of adipose-derived and bone marrow-derived stroma. PLoS One 2012, 7:e30563.
• [23]Spaeth EL, Dembinski JL, Sasser AK, Watson K, Klopp A, Hall B, Andreeff M, Marini F: Mesenchymal Stem Cell Transition to Tumor-Associated Fibroblasts Contributes to Fibrovascular Network Expansion and Tumor Progression. PLoS One 2009, 4:e4992.
• [24]Pasquet M, Golzio M, Mery E, Rafii A, Benabbou N, Mirshahi P, Hennebelle I, Bourin P, Allal B, Teissie J, et al.: Hospicells (ascites-derived stromal cells) promote tumorigenicity and angiogenesis. Int J Cancer 2010, 126:2090-2101.
• [25]McLean K, Gong Y, Choi Y, Deng N, Yang K, Bai S, Cabrera L, Keller E, McCauley L, Cho KR, Buckanovich RJ: Human ovarian carcinoma-associated mesenchymal stem cells regulate cancer stem cells and tumorigenesis via altered BMP production. J Clin Invest 2011, 121:3206-3219.
• [26]Chen J, Yao Y, Gong C, Yu F, Su S, Liu B, Deng H, Wang F, Lin L, Yao H, et al.: CCL18 from tumor-associated macrophages promotes breast cancer metastasis via PITPNM3. Cancer Cell 2011, 19:541-555.
• [27]Valastyan S, Weinberg RA: Tumor metastasis: molecular insights and evolving paradigms. Cell 2011, 147:275-292.
• [28]Grivennikov SI, Greten FR, Karin M: Immunity, inflammation, and cancer. Cell 2010, 140:883-899.
• [29]Rath KS, Funk HM, Bowling MC, Richards WE, Drew AF: Expression of soluble interleukin-6 receptor in malignant ovarian tissue. Am J Obstet Gynecol 2010, 203:230 e231-238.
• [30]Giridhar PV, Funk HM, Gallo CA, Porollo A, Mercer CA, Plas DR, Drew AF: Interleukin-6 receptor enhances early colonization of the murine omentum by upregulation of a mannose family receptor, LY75, in ovarian tumor cells. Clin Exp Metastasis 2011, 28:887-897.
• [31]Hong DS, Angelo LS, Kurzrock R: Interleukin-6 and its receptor in cancer: implications for translational therapeutics. Cancer 2007, 110:1911-1928.
• [32]Taga T, Hibi M, Hirata Y, Yamasaki K, Yasukawa K, Matsuda T, Hirano T, Kishimoto T: Interleukin-6 triggers the association of its receptor with a possible signal transducer, gp130. Cell 1989, 58:573-581.
• [33]Stahl N, Farruggella TJ, Boulton TG, Zhong Z, Darnell JE Jr, Yancopoulos GD: Choice of STATs and other substrates specified by modular tyrosine-based motifs in cytokine receptors. Science 1995, 267:1349-1353.
• [34]Yamanaka Y, Nakajima K, Fukada T, Hibi M, Hirano T: Differentiation and growth arrest signals are generated through the cytoplasmic region of gp130 that is essential for Stat3 activation. EMBO J 1996, 15:1557-1565.
• [35]Wegenka UM, Buschmann J, Lutticken C, Heinrich PC, Horn F: Acute-phase response factor, a nuclear factor binding to acute-phase response elements, is rapidly activated by interleukin-6 at the posttranslational level. Mol Cell Biol 1993, 13:276-288.
• [36]Kidd S, Spaeth E, Dembinski JL, Dietrich M, Watson K, Klopp A, Battula VL, Weil M, Andreeff M, Marini FC: Direct Evidence of Mesenchymal Stem Cell Tropism for Tumor and Wounding Microenvironments Using In Vivo Bioluminescent Imaging. Stem Cells 2009, 27:2614-2623.
• [37]Colomiere M, Ward AC, Riley C, Trenerry MK, Cameron-Smith D, Findlay J, Ackland L, Ahmed N: Cross talk of signals between EGFR and IL-6R through JAK2/STAT3 mediate epithelial-mesenchymal transition in ovarian carcinomas. Br J Cancer 2009, 100:134-144.
• [38]Scheel C, Eaton EN, Li SH, Chaffer CL, Reinhardt F, Kah KJ, Bell G, Guo W, Rubin J, Richardson AL, Weinberg RA: Paracrine and autocrine signals induce and maintain mesenchymal and stem cell states in the breast. Cell 2011, 145:926-940.
• [39]Liu S, Ginestier C, Ou SJ, Clouthier SG, Patel SH, Monville F, Korkaya H, Heath A, Dutcher J, Kleer CG, et al.: Breast cancer stem cells are regulated by mesenchymal stem cells through cytokine networks. Cancer Res 2011, 71:614-624.