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Head and neck squamous cancer stromal fibroblasts produce growth factors influencing phenotype of normal human keratinocytes

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Abstract

Epithelial–mesenchymal interaction between stromal fibroblasts and cancer cells influences the functional properties of tumor epithelium, including the tumor progression and spread. We compared fibroblasts prepared from stroma of squamous cell carcinoma and normal dermal fibroblasts concerning their biological activity toward normal keratinocytes assessed by immunocytochemistry and profiling of gene activation for growth factors/cytokines by microarray chip technology. IGF-2 and BMP-4 were determined as candidate factors responsible for tumor-associated fibroblast activity that influences normal epithelia. This effect was confirmed by addition of recombinant IGF-2 and BMP4, respectively, to the culture medium. This hypothesis was also verified by inhibition experiments where blocking antibodies were employed in the medium conditioned by cancer-associated fibroblast. Presence of these growth factors was also detected in tumor samples.

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References

  • Aberdam D (2004) Derivation of keratinocyte progenitor cells and skin formation from embryonic stem cells. Int J Dev Biol 48:203–206

    Article  CAS  PubMed  Google Scholar 

  • Arciniegas E, Neves YC, Carrillo LM (2006) Potential role for insulin-like growth factor II and vitronectin in the endothelial–mesenchymal transition process. Differentiation 74:277–292

    Article  CAS  PubMed  Google Scholar 

  • Bendall SC, Stewart MH, Menendez P, Georgie D, Vijayaragavan K, Werbowetski-Ogilvie T, Ramos-Mejia V, Rouleau A, Yang J, Bosse M, Lajoie G, Bhatia M (2007) IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent human cells in vitro. Nature 448:1015–1021

    Article  CAS  PubMed  Google Scholar 

  • Brady G, Crean S, Naik P, Kapas S (2007) Upregulation of IGF-2 and IGF-1 receptor expression in oral cancer cell lines. Int J Oncol 31:875–881

    CAS  PubMed  Google Scholar 

  • Casanova ML, Bravo A, Martinez-Palacio J, Fernandez-Acenero MJ, Villanueva C, Larcher F, Conti CJ, Jorcano JL (2004) Epidermal abnormalities and increased malignancy of skin tumors in human epidermal keratin 8-expressing transgenic mice. FASEB J 18:1556–1558

    CAS  PubMed  Google Scholar 

  • De Wever O, Mareel M (2003) Role of tissue stroma in cancer cell invasion. J Pathol 200:429–447

    Article  PubMed  CAS  Google Scholar 

  • Diehl D, Oesterle D, Elmlinger MW, Hoeflich A, Wolf E, Lam H (2006) IGF-II transgenic mice display increased aberrant colon crypt multiplicity and tumor volume after 1, 2 dimethylhydrazine treatment. J Karcinogen 5:24

    Article  CAS  Google Scholar 

  • Dunning M, Smith M, Camilier I, Ritchie M (2007) Beadarray: quality control and low-level analysis of Beadarrays. R package version 1.6.0

  • Durinck S, Moreau Y, Kasprzyk A, Davis S, De Moor B, Brazma A, Huber W (2005) BioMart and Bioconductor: a powerful link between biological databases and microarray data analysis. Bioinformatics 21:3439–3440

    Article  CAS  PubMed  Google Scholar 

  • Dvorak HF (1986) Tumors: wounds that do not heal. New Engl J Med 315:1650–1689

    Article  CAS  PubMed  Google Scholar 

  • Ellis JM, Jenkins S, Hanfelt J, Redington ME, Taylor M, Leek R, Siddle K, Harris A (1998) Insulin-like growth factors in human breast cancer. Breast Cancer Res Treat 52:175–184

    Article  CAS  PubMed  Google Scholar 

  • Froňková V, Holíková Z, Liu F-T, Homolka J, Rijken D-C, André S, Bovin NV, Smetana K Jr, Gabius H-J (1999) Simultaneous detection of endogenous lectins and their binding capability at the single cell-level-a technical note. Folia Biol 45:157–162

    Google Scholar 

  • Fuchs E (2007) Scratching the surface of skin development. Nature 445:834–842

    Article  CAS  PubMed  Google Scholar 

  • García-Echeverría C, Pearson MA, Marti A, Meyer T, Mestan J, Zimmermann J, Gao J, Brueggen J, Capraro H-G, Cozens R, Evans DB, Fabbro D, Furet P, Graus Porta D, Liebetanz J, Martiny-Baron G, Ruetz S, Hofmann F (2004) In vivo antitumor activity of NVP-AEW541-A novel, potent, and selective inhibitor of the IGF-IR dinase. Cancer Cell 5:231–239

    Article  PubMed  Google Scholar 

  • Gires O, Menz M, Schaffrik M, Kleu C, Rauch J, Ahlemann M, Berle D, Mack B, Wolleneberg B, Lang S, Hofmann T, Hammerschmidt W, Zeidler R (2004) Profile identification of disease-associated humoral antigens using AMIDA, a novel proteomics-based technology. Cell Mol Life Sci 61:1198–1207

    Article  CAS  PubMed  Google Scholar 

  • Grando SA, Schofield O, Skubitz APN, Kist DA, Zelickson BD, Zachary CB (1996) Nodular basal cell carcinomain vivo vs in vitro. Arch Dermatol 132:1185–1193

    Article  CAS  PubMed  Google Scholar 

  • Grbesa I, Ivkic M, Pegan B, Gall-Troselj K (2006) Loss of imprinting and promoter usage of the IGF2 in laryngeal squamous cell carcinoma. Cancer Lett 238:224–229

    Article  CAS  PubMed  Google Scholar 

  • Grinnell KL, Bickenbach JR (2007) Skin keratinocytes pre-treated with embryonic stem cell-conditioned medium or BMP4 can be directed to an alternative cell lineages. Cell Prolif 40:685–705

    Article  CAS  PubMed  Google Scholar 

  • Hamada S, Satoh K, Hirota M, Kimura K, Kanno A, Masamune A, Shimosegawa T (2007) Bone morphogenetic protein 4 induces epithelial-mesenchymal transition through MSX2 induction on pancreatic cancer cell line. J Cell Physiol 213:768–774

    Article  CAS  PubMed  Google Scholar 

  • Hawsawi NM, Ghebeh H, Hendrayani SF, Tulbah A, Al-Eid M, Al-Tweigeri T, Ajarim D, Alaiya A, Dermime S, Aboussekhra A (2008) Breast carcinoma-associated fibroblasts and their counterparts display neoplastic-specific changes. Cancer Res 68(8):2717–2725

    Article  CAS  PubMed  Google Scholar 

  • Hwang RF, Moore T, Arumugam T, Ramachandran V, Amos KD, Rivera A, Ji B, Evans DB, Logsdon CD (2008) Cancer-associated stromal fibroblasts promote pancreatic tumor progression. Cancer Res 68:918–926

    Google Scholar 

  • Jelinic P, Shaw P (2007) Loss of imprinting and cancer. J Pathol 211:261–268

    Article  CAS  PubMed  Google Scholar 

  • Jin Y, Tipoe GL, Liong EC, Lau TYH, Fung PCW, Lejny KM (2001) Overexpression of BMP-2/4, -5, and BMPR-IA associated with malignancy of oral epithelium. Oral Oncol 37:225–233

    Article  PubMed  Google Scholar 

  • Kaminski A, Hahne JC, Hadouti EM, Florin A, Wellmann A, Wernert N (2006) Tumour-stroma interactions between metastatic prostate cancer cells and fibroblasts. Int J Mol Med 18:941–950

    CAS  PubMed  Google Scholar 

  • Karafiat A, Dvořákova M, Krejci E, Kralova J, Pajera P, Snajdr P, Mandikova S, Bartunek P, Grim M, Dvorak M (2005) Transcription factor c-Myb is involved in the regulation of the epithelial-mesenchymal transition in the avian neural crest. Cell Mol Life Sci 62:2516–2525

    Article  CAS  PubMed  Google Scholar 

  • Klíma J, Lacina L, Dvořánková B, Herrmann D, Carnwath JW, Niemann H, Kaltner H, André S, Motlík J, Gabius H-J, Smetana K Jr (2009) Differential regulation of galectin expression/reactivity during wound healing in porcine skin and in cultures of epidermal cells with functional impact on migration. Physiol Res (in press)

  • Kornprat P, Rehak P, Rüschoff J, Langner C (2006) Expression of IGF-I, IGF-II, and IGF-IR in gallbladder carcinoma. A systematic analysis including primary and corresponding metastatic tumours. J Clin Pathol 59:202–206

    Article  CAS  PubMed  Google Scholar 

  • Koukourakis MI, Giatromanolaki A, Hartus AL, Sivridis E (2006) Comparison of metabolic pathways between cancer cells and stromal cells in colorectal carcinomas: a metabolic survival role for tumor-associated stroma. Cancer Res 66:632–637

    Article  CAS  PubMed  Google Scholar 

  • Lacina L, Smetana K Jr, Dvořánková B, Štork J, Plzáková Z, Gabius H-J (2006) Immunocyto- and histochemical profiling of nucleostemin expression: marker of epidermal stem cells? J Dermatol Sci 44:73–80

    Article  CAS  PubMed  Google Scholar 

  • Lacina L, Dvořánkova B, Smetana K Jr, Chovanec M, Plzák J, Tachezy R, Kideryová L, Kučerová L, Čada Z, Bouček J, Kodet R, André S, Gabius H-J (2007a) Marker profiling of normal keratinocytes identifies the stroma from squamous cell carcinoma of the oral cavity as a modulatory microenvironment in co-culture. Int J Radiation Biol 83:837–848

    Article  CAS  Google Scholar 

  • Lacina L, Smetana K Jr, Dvořánková B, Pytlík R, Kideryová L, Kučerová L, Plzáková Z, Štork J, Gabius H-J, André S (2007b) Stromal fibroblasts from basal cell carcinoma affect phenotype of normal keratinocytes. Br J Dermatol 156:819–829

    Article  CAS  PubMed  Google Scholar 

  • Li L, Neaves WB (2006) Normal stem cells and cancer stem cells: the niche matters. Cancer Res 66:4553–4557

    Google Scholar 

  • Lorusso G, Rüegg C (2008) The tumor microenvironment and its contribution to tumor evolution toward metastasis. Histochem Cell Biol 130(6):1091–1103

    Google Scholar 

  • Mathur SP, Mathur RS, Underwood PB, Kohler MF, Creasman WT (2003) Circulating levels of insulin-like growth factor-II and IGF-binding protein 3 in cervical cancer. Gyn Oncol 91:486–493

    Article  CAS  Google Scholar 

  • Musselmann K, Kane BP, Alexandrou B, Hassell JR (2008) IGF-II is present in bovine corneal stroma and activates keratocytes to proliferate in vitro. Exp Eye Res 86:506–511

    Article  CAS  PubMed  Google Scholar 

  • Nosho K, Yamamoto H, Adachi Y, Endo T, Hinoda Y, Imai K (2005) Gene expression profiling of colorectal adenomas and early invasive carcinomas by cDNA array analysis. Br J Cancer 92:1193–1200

    Article  CAS  PubMed  Google Scholar 

  • Orimo A, Gupta PB, Sgroi DC, Arenzana-Seisdedos F, Delaunay T, Naeem R, Carey VJ, Richardson AL, Weinberg RA (2005) Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. Cell 121:335–348

    Article  CAS  PubMed  Google Scholar 

  • Pavelić J, Radaković B, Pavelić K (2007) Insulin-like growth factor 2 and its receptors (IGF 1R and IGF 2R/manose 6-phosphate) in endometrial adenocarcinoma. Gyn Oncol 105:727–735

    Article  CAS  Google Scholar 

  • Potenta S, Zeisberg E, Kalluri R (2008) The role of endothelial-to mesenchymal transition in cancer progression. Br J Cancer 99(9):1375–1379

    Google Scholar 

  • R Development Core Team R (2007) A language and environment for statistical computing. R Foundation for Statistical Computing, Vienna, Austria. ISBN 3-900051-07-0, URL http://www.R-project.org

  • Rheinwald JG, Green H (1975) Serial cultivation of strains of human epidermal keratinocytes; the formation of keratinizing colonies from the single cells. Cell 6:331–344

    Article  CAS  PubMed  Google Scholar 

  • Rothhammer T, Poser I, Soncin F, Bataille F, Moser M, Bosserhoff AK (2005) Bone morphogenic proteins are overexpressed in malignant melanoma and promote cell invasion and migration. Cancer Res 65:448–456

    CAS  PubMed  Google Scholar 

  • Sayer RA, Lancaster JM, Pittmanc J, Grayd J, Whitaker R, Markse JR, Berchuckb A (2005) High insulin-like growth factor-2 (IGF-2) gene expression is an independent predictor of poor survival for patients with advanced stage serous epithelial ovarian cancer. Gynecol Oncol 96:355–361

    Article  CAS  PubMed  Google Scholar 

  • Sell S (2004) Stem cell origin of cancer and differentiation therapy. Crit Rev Oncol Hematol 51:1–28

    Article  PubMed  Google Scholar 

  • Smyth GK (2005) Limma: linear models for microarray data. In: Gentleman R, Carey V, Dudoit S, Irizarry R, Huber W (eds) Bioinformatics and computational biology solutions using R and bioconductor. Springer, New York, pp 397–420

    Chapter  Google Scholar 

  • Thériault BL, Shepherd TG, Mujoomdar ML, Nachtigal MW (2007) BMP4 induces EMT and Rho GTPase activation in human ovarian cancer cells. Carcinogenesis 28:1153–1162

    Article  PubMed  CAS  Google Scholar 

  • Thiery JP, Sleeman JP (2006) Complex networks orchestrace epithelial-mesenchymal transitions. Nature Rev Mol Cel Biol 7(2):131–142

    Google Scholar 

  • Trojan L, Bode C, Weiss C, Mayer D, Grobholz R, Alken P, Michel MS (2006) IGF-II serum levels increase discrimination between benign prostatic hyperplasia and prostate cancer and improve the predictive value of PSA in clinical staging. Eur Urol 49:286–292

    Article  CAS  PubMed  Google Scholar 

  • Troy T-C, Turkmen K (2005) Commitment of embryonic stem cells to an epidermal cell fate and differentiation. Dev Dynam 232:293–300

    Article  CAS  Google Scholar 

  • Vandesompele J, De Preter K, Pattyn F, Poppe B, Van Roy N, De Paepe A, Speleman F (2002) Accurate normalization of real-time quantitative RT-PCR data by geometric averaging of multiple internal control genes. Genome Biol 18:3(7)

    Google Scholar 

  • Weber A, Hengge UR, Stricker IS, Tischoff I, Markwart A, Anhalt K, Dietz A, Wittekind C, Tannapfel A (2007) Protein microarrays for the detection of biomarkers in head and neck squamous cell carcinomas. Hum Pathol 38:228–238

    Article  CAS  PubMed  Google Scholar 

  • Wilson PA, Hemmati-Brivanlou A (1995) Induction of epidermis and inhibition of neural fate by Bmp-4. Nature 376:331–333

    Article  CAS  PubMed  Google Scholar 

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Acknowledgments

This study was supported by Ministry of Education, Youth and Sport of the Czech Republic, projects No. MSM0021620806, NPVII 2B06106, AV0Z50520514 and project of Grant Agency of the Czech Republic No. 304/08/P175. Authors are grateful to Iva Burdová and Vít Hajdúch for excellent technical assistance.

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Authors and Affiliations

  1. Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, Vídeňská 1083, 142 20, Prague 4, Czech Republic

    Hynek Strnad, Michal Kolář, Čestmír Vlček, Jana Šáchová & Markéta Urbanová

  2. Department of Dermatovenereology, Institute of Anatomy, First Faculty of Medicine, Charles University in Prague, U Nemocnice 2, 128 00, Prague 2, Czech Republic

    Lukáš Lacina

  3. Institute of Anatomy, First Faculty of Medicine, Charles University in Prague, U Nemocnice 3, 128 00, Prague 2, Czech Republic

    Barbora Dvořánková & Karel Smetana Jr

  4. Department of Otorhinolaryngology, Head and Neck Surgery, First Faculty of Medicine, Charles University in Prague, V Úvalu 84, 150 06, Prague 5, Czech Republic

    Jan Betka & Markéta Urbanová

  5. Department of Otorhinolaryngology, Head and Neck Surgery, Institute of Anatomy, First Faculty of Medicine, Charles University in Prague, V Úvalu 84, 150 06, Prague 5, Czech Republic

    Zdeněk Čada, Jan Plzák & Martin Chovanec

  6. Department of Stomatology, First Faculty of Medicine, Charles University in Prague, Kateřinská 32, 128 01, Prague 2, Czech Republic

    Jaroslav Valach

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  1. Hynek Strnad
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Correspondence to Karel Smetana Jr.

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Strnad, H., Lacina, L., Kolář, M. et al. Head and neck squamous cancer stromal fibroblasts produce growth factors influencing phenotype of normal human keratinocytes. Histochem Cell Biol 133, 201–211 (2010). https://doi.org/10.1007/s00418-009-0661-6

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