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Interleukin-6 promotes human epidermal keratinocyte proliferation and keratin cytoskeleton reorganization in culture

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Abstract

We have studied the effects of interleukin-6 (IL-6) on human epidermal keratinocytes by using serum-free culture conditions that allow the serial transfer, differentiation, and formation of well-organized multilayered epithelia. IL-6 at 2.5 ng/ml or higher concentrations promoted keratinocyte proliferation, with an ED50 of about 15 ng/ml and a maximum effect at 50 ng/ml. IL-6 was 10-fold less potent than epidermal growth factor (EGF) or transforming growth factor-α (TGF-α) and supported keratinocyte growth for up to eight cumulative cell generations. IL-6-treated keratinocytes formed highly stratified colonies with a narrower proliferative/migratory rim than those keratinocytes stimulated with EGF or TGF-α; confluent epithelial sheets treated with IL-6 also underwent an increase in the number of cell layers. We also examined the effect of IL-6 on the keratin cytoskeleton. Immunostaining with anti-K16 monoclonal antibodies showed that the keratin network was aggregated and reorganized around cell nucleus and that this was not attributable to changes in keratin levels. This is the first report concerning the induction of the reorganization of keratin intermediate filaments by IL-6 in human epidermal keratinocytes.

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References

  • Akira S, Taga T, Kishimoto T (1993) Interleukin-6 in biology and medicine. Adv Immunol 54:1–78

    Article  PubMed  CAS  Google Scholar 

  • Albanesi C, Scarponi C, Cavani A, Federici M, Nasorri F, Girolomoni G (2000) Interleukin-17 is produced by both Th1 and Th2 lymphocytes, and modulates interferon-gamma- and interleukin-4-induced activation of human keratinocytes. J Invest Dermatol 115:81–87

    Article  PubMed  CAS  Google Scholar 

  • Aragane Y, Yamada H, Schwarz A, Poppelmann B, Luger TA, Tezuka T, Schwarz T (1996) Transforming growth factor-alpha induces interleukin-6 in the human keratinocyte cell line HaCaT mainly by transcriptional activation. J Invest Dermatol 106:1192–1197

    Article  PubMed  CAS  Google Scholar 

  • Asselineau D, Bernhard B, Bailly C, Darmon M (1985) Epidermal morphogenesis and induction of the 67kd keratin polypeptide of human keratinocytes at the liquid-air interface. Exp Cell Res 159:536–539

    Article  PubMed  CAS  Google Scholar 

  • Barrandon Y, Green H (1987) Cell migration is essential for sustained growth of keratinocyte colonies: the roles of transforming growth factor-alpha and epidermal growth factor. Cell 50:1131–1137

    Article  PubMed  CAS  Google Scholar 

  • Bechtel MJ, Wysocki NS, Heidtmann A, Stark HJ, Fusenig N, Kramer MD, Schaefer BM (1998) Plasminogen activator inhibitor type 2 is expressed in keratinocytes during re-epithelialization of epidermal defects. Br J Dermatol 138:22–28

    Article  PubMed  CAS  Google Scholar 

  • Bolívar-Flores J, Poumian E, Marsch-Moreno M, Montes de Oca G, Kuri-Harcuch W (1990) Use of cultured human epidermal keratinocytes for allografting burns and conditions for temporary banking of the cultured allografts. Burns 16:3–8

    Article  PubMed  Google Scholar 

  • Boxman I, Lowik C, Aarden L, Ponec M (1993) Modulation of IL-6 production and IL-1 activity by keratinocyte-fibroblast interaction. J Invest Dermatol 101:316–324

    Article  PubMed  CAS  Google Scholar 

  • Bujia J, Kim C, Ostos P, Kastenbauer E, Hultner L (1996) Role of interleukin 6 in epithelial hyperproliferation and bone resorption in middle ear cholesteatomas. Eur Arch Otorhinolaryngol 253:152–157

    Article  PubMed  CAS  Google Scholar 

  • Bussolati B, Mariano F, Biancone L, Foa R, David S, Cambi V, Camussi G (1999) Interleukin-12 is synthesized by mesangial cells and stimulates platelet-activating factor synthesis, cytoskeletal reorganization, and cell shape change. Am J Pathol 154:623–632

    PubMed  CAS  Google Scholar 

  • Camargo CA, Madden JF, Gao W, Selvan RS, Clavien PA (1997) Interleukin-6 protects liver against warm ischemia/reperfusion injury and promotes hepatocyte proliferation in the rodent. Hepatology 26:1513–1520

    Article  PubMed  CAS  Google Scholar 

  • Castell JV, Gomez-Lechon MJ, David M, Andus T, Geiger T, Trullenque R, Fabra R, Heinrich PC (1989) Interleukin-6 is the major regulator of acute phase protein synthesis in adult human hepatocytes. FEBS Lett 242:237–239

    Article  PubMed  CAS  Google Scholar 

  • Castell-Rodellas A, Castell JV, Ramirez-Bosca A, Nicolas JF, Valcuende-Cavero F, Thivolet J (1992) Interleukin-6 in normal skin and psoriasis. Acta Derm Venereol 72:165–168

    Google Scholar 

  • Castro-Muñozledo F, Hernández-Quintero M, Marsch-Moreno M, Kuri-Harcuch W (1997a) Cultivation serial transfer and differentiation of epidermal keratinocytes in serum-free medium. Biochem Biophys Res Commun 236:167–172

    Article  PubMed  Google Scholar 

  • Castro-Muñozledo F, Valencia-García C, Kuri-Harcuch W (1997b) Cultivation of rabbit corneal epithelial cells in serum-free medium. Invest Ophthalmol Vis Sci 38:2234–2244

    PubMed  Google Scholar 

  • Chou CF, Omary MB (1993) Mitotic arrest-associated enhancement of O-linked glycosylation and phosphorylation of human keratins 8 and 18. J Biol Chem 268:4465–4472

    PubMed  CAS  Google Scholar 

  • Coulie PG, Stevens M, Van Snick J (1989) High- and low-affinity receptors for murine interleukin 6. Distinct distribution on B and T cells. Eur J Immunol 19:2107–2114

    Article  PubMed  CAS  Google Scholar 

  • Cumberbatch M, Dearman RJ, Kimber I (1996) Constitutive and inducible expression of interleukin-6 by Langerhans cells and lymph node dendritic cells. Immunology 87:513–518

    Article  PubMed  CAS  Google Scholar 

  • DeFife KM, Jenney CR, Colton E, Anderson JM (1999) Cytoskeletal and adhesive structural polarizations accompany IL-13-induced human macrophage fusion. J Histochem Cytochem 47:65–74

    PubMed  CAS  Google Scholar 

  • DiColandrea T, Karashima T, Määttäa A, Watt FM (2000) Subcellular distribution of envoplakin and periplakin: insights into their role as precursors of the epidermal cornified envelope. J Cell Biol 151:573–585

    Article  PubMed  CAS  Google Scholar 

  • Eichner R, Bonitz P, Sun TT (1984) Classification of epidermal keratins according to their immunoreactivity, isoelectric point, and mode of expression. J Cell Biol 98:1388–1396

    Article  PubMed  CAS  Google Scholar 

  • Fiorillo MT, Cabibbo A, Iacopetti P, Fattori E, Ciliberto G (1992) Analysis of human/mouse interleukin-6 hybrid proteins: both amino and carboxy termini of human interleukin-6 are required for in vitro receptor binding. Eur J Immunol 22:2609–2615

    Article  PubMed  CAS  Google Scholar 

  • Freedberg IM, Tomic-Canic M, Komine M, Blumenberg M (2001) Keratins and the keratinocyte activation cycle. J Invest Dermatol 116:633–640

    Article  PubMed  CAS  Google Scholar 

  • Gallucci RM, Simeonova PP, Matheson JM, Kommineni C, Guriel JL, Sugawara T, Luster MI (2000) Impaired cutaneous wound healing in interleukin-6-deficient and immunosuppressed mice. FASEB J 14:2525–2531

    Article  PubMed  CAS  Google Scholar 

  • Gallucci RM, Sloan DK, Heck JM, Murray AR, O’Dell SJ (2004) Interleukin 6 indirectly induces keratinocyte migration. J Invest Dermatol 122:764–772

    Article  PubMed  CAS  Google Scholar 

  • Grossman RM, Krueger J, Yourish D, Granelli-Piperno A, Murphy DP, May LT, Kupper TS, Sehgal PB, Gottlieb AB (1989) Interleukin 6 is expressed in high levels in psoriatic skin and stimulates proliferation of cultured human keratinocytes. Proc Natl Acad Sci USA 86:6367–6371

    Article  PubMed  CAS  Google Scholar 

  • Hancock GE, Kaplan G, Cohn ZA (1988) Keratinocyte growth regulation by the products of immune cells. J Exp Med 168:1395–1402

    Article  PubMed  CAS  Google Scholar 

  • Hattori N, Komine M, Yano S, Kaneko T, Hanakawa Y, Hashimoto K, Tamaki K (2002) Interferon-gamma, a strong suppressor of cell proliferation, induces upregulation of keratin K6, one of the inflammatory- and proliferation-associated keratins. J Invest Dermatol 119:403–410

    Article  PubMed  CAS  Google Scholar 

  • Heinrich PC, Behrmann I, Haan S, Hermanns HM, Muller-Newen G, Schaper F (2003) Principles of interleukin (IL)-6-type cytokine signalling and its regulation. Biochem J 374:1–20

    Article  PubMed  CAS  Google Scholar 

  • Hennings H, Michael D, Cheng C, Steinert P, Holbrook K, Yuspa SH (1980) Calcium regulation of growth and differentiation of mouse epidermal cells in culture. Cell 19:245–254

    Article  PubMed  CAS  Google Scholar 

  • Hilbert DM, Kopf M, Mock BA, Köhler G, Rudikoff S (1995) Interleukin 6 is essential for in vivo development of B lineage neoplasms. J Exp Med 182:243–248

    Article  PubMed  CAS  Google Scholar 

  • Horii Y, Muraguchi A, Iwano M, Matsuda T, Hirayama T, Yamada H, Fujii Y, Dohi K, Ishikawa H, Ohmoto Y (1989) Involvement of IL-6 in mesangial proliferative glomerulonephritis. J Immunol 143:3949–3955

    PubMed  CAS  Google Scholar 

  • Jiang CK, Magnaldo T, Ohtsuki M, Freedberg IM, Bernerd F, Blumenberg M (1993) Epidermal growth factor and transforming growth factor alpha specifically induce the activation- and hyperproliferation-associated keratins 6 and 16. Proc Natl Acad Sci USA 90:6786–6790

    Article  PubMed  CAS  Google Scholar 

  • Jorissen RN, Walker F, Pouliot N, Garrett TP, Ward CW, Burgess AW (2003) Related epidermal growth factor receptor: mechanisms of activation and signalling. Exp Cell Res 284:31–53

    Article  PubMed  CAS  Google Scholar 

  • Kammüller ME (1995) Recombinant human interleukin-6: safety issues of a pleiotropic growth factor. Toxicology 105:91–107

    Article  PubMed  Google Scholar 

  • Klymkowsky MW, Maynell LA, Nislow C (1991) Cytokeratin phosphorylation, cytokeratin filament severing and the solubilization of the maternal mRNA Vg1. J Cell Biol 114:787–797

    Article  PubMed  CAS  Google Scholar 

  • Komine M, Freedberg IM, Blumenberg M (1996) Regulation of epidermal expression of keratin K17 in inflammatory skin diseases. J Invest Dermatol 107:569–575

    Article  PubMed  CAS  Google Scholar 

  • Komine M, Rao LS, Freedberg IM, Simon M, Milisavljevic V, Blumenberg M (2001) Interleukin-1 induces transcription of keratin K6 in human epidermal keratinocytes. J Invest Dermatol 116:330–338

    Article  PubMed  CAS  Google Scholar 

  • Kopan R, Traska G, Fuchs E (1987) Retinoids as important regulators of terminal differentiation: examining keratin expression in individual epidermal cells at various stages of keratinization. J Cell Biol 105:427–440

    Article  PubMed  CAS  Google Scholar 

  • Krueger J, Ray A, Tamm I, Sehgal PB (1991) Expression and function of interleukin-6 in epithelial cells. J Cell Biochem 45:327–334

    Article  PubMed  CAS  Google Scholar 

  • Markey AC, Lane EB, MacDonald DM, Leigh IM (1992) Keratin expression in basal cell carcinomas. Br J Dermatol 126:154–160

    Article  PubMed  CAS  Google Scholar 

  • Mazzalupo S, Wong P, Martin P, Coulombe PA (2003) Role for keratins 6 and 17 during wound closure in embryonic mouse skin. Dev Dyn 226:356–365

    Article  PubMed  CAS  Google Scholar 

  • Miki S, Iwano M, Miki Y, Yamamoto M, Tang B, Yokokawa K, Sonoda T, Hirano T, Kishimoto T (1989) Interleukin-6 (IL-6) functions as an in vitro autocrine growth factor in renal cell carcinomas. FEBS Lett 250:607–610

    Article  PubMed  CAS  Google Scholar 

  • Mouawad R, Benhammouda A, Rixe O, Antoine EC, Borel C, Weil M, Khayat D, Soubrane C (1996) Endogenous interleukin 6 levels in patients with metastatic malignant melanoma: correlation with tumor burden. Clin Cancer Res 2:1405–1409

    PubMed  CAS  Google Scholar 

  • Nakatani S, Okada N, Okumura H, Yoshikawa K (1992) A two-colour flow-cytometric study of cell kinetics and differentiation of human keratinocytes in culture. Arch Dermatol Res 284:349–352

    Article  PubMed  CAS  Google Scholar 

  • Neuner P, Urbanski A, Trautinger F, Möller A, Kirnbauer R, Kapp A, Schöpf E, Schwarz T, Luger TA (1991) Increased IL-6 production by monocytes and keratinocytes in patients with psoriasis. J Invest Dermatol 97:27–33

    Article  PubMed  CAS  Google Scholar 

  • Nishida T, Nakamura M, Mishima H, Otori T (1992a) Interleukin 6 promotes epithelial migration by a fibronectin-dependent mechanism. J Cell Physiol 153:1–5

    Article  PubMed  CAS  Google Scholar 

  • Nishida T, Nakamura M, Mishima H, Otori T, Hikida M (1992b) Interleukin 6 facilitates corneal epithelial wound closure in vivo. Arch Ophthalmol 110:1292–1294

    PubMed  CAS  Google Scholar 

  • O’Guin WM, Schermer A, Sun TT (1985) Immunofluorescence staining of keratin filaments in cultured epithelial cells. J Tissue Culture Methods 9:123–128

    Article  Google Scholar 

  • Paladini RD, Takahashi K, Bravo NS, Coulombe PA (1996) Onset of reepithelialization after skin injury correlates with a reorganization of keratin filaments in wound edge keratinocytes: defining a potential role of keratin 16. J Cell Biol 132:381–397

    Article  PubMed  CAS  Google Scholar 

  • Poumay Y, Pittelkow MR (1995) Cell density and culture factors regulate keratinocyte commitment to differentiation and expression of suprabasal K1/K10 keratins. J Invest Dermatol 104:271–276

    Article  PubMed  CAS  Google Scholar 

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

    Article  PubMed  CAS  Google Scholar 

  • Ruef C, Budde K, Lacy J, Northemann W, Baumann M, Sterzel RB, Coleman DL (1990) Interleukin 6 is an autocrine growth factor for mesangial cells. Kidney Int 38:249–257

    Article  PubMed  CAS  Google Scholar 

  • Sato M, Sawamura D, Ina S, Yaguchi T, Hanada K, Hashimoto I (1999) In vivo introduction of the interleukin 6 gene into human keratinocytes: induction of epidermal proliferation by the fully spliced form of interleukin 6, but not by the alternatively spliced form. Arch Dermatol Res 291:400–404

    Article  PubMed  CAS  Google Scholar 

  • Sawamura D, Meng X, Ina S, Sato M, Tamai K, Hanada K, Hashimoto I (1998) Induction of keratinocyte proliferation and lymphocytic infiltration by in vivo introduction of the IL-6 gene into keratinocytes and possibility of keratinocyte gene therapy for inflammatory skin diseases using IL-6 mutant genes. J Immunol 161:5633–5639

    PubMed  CAS  Google Scholar 

  • Shipley GD, Keeble WW, Hendrickson JE, Coffey RJ, Pittelkow MR (1989) Growth of normal human keratinocytes and fibroblasts in serum-free medium is stimulated by acidic and basic fibroblast growth factor. J Cell Physiol 138:511–518

    Article  PubMed  CAS  Google Scholar 

  • Strnad P, Windoffer R, Leube RE (2001) In vivo detection of cytokeratin filament network breakdown in cells treated with the phosphatase inhibitor okadaic acid. Cell Tissue Res 306:277–293

    Article  PubMed  CAS  Google Scholar 

  • Strnad P, Windoffer R, Leube RE (2002) Induction of rapid and reversible cytokeratin filament network remodeling by inhibition of tyrosine phosphatases. J Cell Sci 115:4133–4148

    Article  PubMed  CAS  Google Scholar 

  • Sugawara T, Gallucci RM, Simeonova PP, Luster MI (2001) Regulation and role of interleukin 6 in wounded human epithelial keratinocytes. Cytokine 15:328–336

    Article  PubMed  CAS  Google Scholar 

  • Suto N, Ikura K, Sasaki R (1993) Expression induced by interleukin-6 of tissue-type transglutaminase in human hepatoblastoma HepG2 cells. J Biol Chem 268:7469–7473

    PubMed  CAS  Google Scholar 

  • Takahashi K, Folmer J, Coulombe PA (1994) Increased expression of keratin 16 causes anomalies in cytoarchitecture and keratinization in transgenic mouse skin. J Cell Biol 127:505–520

    Article  PubMed  CAS  Google Scholar 

  • Tamm I, Cardinale I, Krueger J, Murphy JS, May LT, Sehgal PB (1989) Interleukin 6 decreases cell-cell association and increases motility of ductal breast carcinoma cells. J Exp Med 170:1649–1669

    Article  PubMed  CAS  Google Scholar 

  • Tamm I, Cardinale I, Kikuchi T, Krueger JG (1994) E-cadherin distribution in interleukin 6-induced cell-cell separation of ductal breast carcinoma cells. Proc Natl Acad Sci USA 91:4338–4342

    Article  PubMed  CAS  Google Scholar 

  • Turksen K, Kupper T, Degenstein L, Williams I, Fuchs E (1992) Interleukin 6: insights to its function in skin by overexpression in transgenic mice. Proc Natl Acad Sci USA 89:5068–5072

    Article  PubMed  CAS  Google Scholar 

  • Van Dam M, Müllberg J, Schooltink H, Stoyan T, Brakenhoff JPJ, Graeve L, Heinrich PC, Rose-John S (1993) Structure-function analysis of interleukin-6 utilizing human/murine chimeric molecules. Involvement of two separate domains in receptor binding. J Biol Chem 268:15285–15290

    PubMed  Google Scholar 

  • Vijayalakshmi V, Gupta PD (1994) Estradiol-regulated transamidation of keratins by vaginal epithelial cell transglutaminase. Exp Cell Res 214:358–366

    Article  PubMed  CAS  Google Scholar 

  • Waelti ER, Sven P, Inaebnit SP, Hans P, Rast HP, Hunziker T, Limat A, Lasse R, Braathen LR, Wiesmann U (1992) Co-culture of human keratinocytes on post-mitotic human dermal fibroblast feeder cells: production of large amounts of interleukin 6. J Invest Dermatol 98:805–808

    Article  PubMed  CAS  Google Scholar 

  • Wang Z, Newman WH (2003) Smooth muscle cell migration stimulated by interleukin 6 is associated with cytoskeletal reorganization. J Surg Res 111:261–266

    Article  PubMed  CAS  Google Scholar 

  • Watt FM (1984) Selective migration of terminally differentiating cells from the basal layer of cultured human epidermis. J Cell Biol 98:16–21

    Article  PubMed  CAS  Google Scholar 

  • Watt FM, Green H (1982) Stratification and terminal differentiation of cultured epidermal cells. Nature 295:434–436

    Article  PubMed  CAS  Google Scholar 

  • Wawersik M, Coulombe PA (2000) Forced expression of keratin 16 alters the adhesion, differentiation and migration of mouse skin keratinocytes. Mol Biol Cell 11:3315–3327

    PubMed  CAS  Google Scholar 

  • Wawersik M, Mazzalupo S, Nguyen D, Coulombe PA (2001) Increased levels of keratin 16 alter epithelialization potential of mouse skin keratinocytes in vivo and ex vivo. Mol Biol Cell 12:3439–3450

    PubMed  CAS  Google Scholar 

  • Wetzels RH, Kuijpers HJ, Lane EB, Leigh IM, Troyanovsky SM, Holland R, Haelst UJ van, Ramaekers FC (1991) Basal cell-specific and hyperproliferation-related keratins in human breast cancer. Am J Pathol 138:751–763

    PubMed  CAS  Google Scholar 

  • Yano T, Tokui T, Nishi Y, Nishizawa K, Shibata M, Kikuchi K, Tsuiki S, Yamauchi T, Inagaki M (1991) Phosphorylation of keratin intermediate filaments by protein kinase C, by calmodulin-dependent protein kinase and by cAMP-dependent protein kinase. Eur J Biochem 197:281–290

    Article  PubMed  CAS  Google Scholar 

  • Yoshizaki K, Nishimoto N, Matsumoto K, Tagoh H, Taga T, Deguchi Y, Kuritani T, Hirano T, Hashimoto K, Okada N, Kishimoto T (1990) Interleukin 6 and expression of its receptor on epidermal keratinocytes. Cytokine 2:381–387

    Article  PubMed  CAS  Google Scholar 

  • Zhu AJ, Watt FM (1996) Expression of a dominant negative cadherin mutant inhibits proliferation and stimulates differentiation of human epidermal keratinocytes. J Cell Sci 109:3013–3023

    PubMed  CAS  Google Scholar 

  • Zhu AJ, Watt FM (1999) β-Catenin signalling modulates proliferative potential of human epidermal keratinocytes independently of intercellular adhesion. Development 126:2285–2298

    PubMed  CAS  Google Scholar 

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Acknowledgements

We are grateful to Meytha Marsch-Moreno (IBQ) for preparing the human epidermal keratinocyte banks and the mytomycin-C-treated feeder cells, to Mrs. Erika Sánchez Guzmán and Ms. Ana Ruth Cristóbal-Ramos for technical assistance, to Ms. Maria Elena Rojano for secretarial assistance, and to Mr. Alberto Rodriguez and Mrs. Columba Guadarrama for their technical assistance in the preparation of all materials.

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Correspondence to Federico Castro-Muñozledo.

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This work was supported in part by CONACyT grant nos. 1314P-N9507 and G28272-N.

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Hernández-Quintero, M., Kuri-Harcuch, W., González Robles, A. et al. Interleukin-6 promotes human epidermal keratinocyte proliferation and keratin cytoskeleton reorganization in culture. Cell Tissue Res 325, 77–90 (2006). https://doi.org/10.1007/s00441-006-0173-9

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