ROLE OF FIBROBLASTS IN ORGANOTYPIC CULTURES OF IMMORTALIZED HUMAN ORAL KERATINOCYTES

섬유모세포의 종류에 따른 불멸화된 구강 각화세포의 삼차원적 배양에 관한 연구

  • Cheong, Jeong-Kwon (Dept. of Oral & Maxillofacial Surgery, Sanggye Paik Hospital, College of Medicine, Inje University) ;
  • Yoon, Kyu-Ho (Dept. of Oral & Maxillofacial Surgery, Sanggye Paik Hospital, College of Medicine, Inje University) ;
  • Kim, Eun-Cheol (Dept. of Oral Pathology, College of Dentistry, Wonkwang University)
  • 정정권 (인제대학교 의과대학 상계백병원 구강악안면외과) ;
  • 윤규호 (인제대학교 의과대학 상계백병원 구강악안면외과) ;
  • 김은철 (원광대학교 치과대학 구강병리학교실)
  • Published : 2007.06.30

Abstract

Objective: In organotypic culture of immortalized human oral keratinocytes (IHOK), the change of the growth and differentiation was investigated according to the fibroblast type and the involvement of mitogen-activated protein (MAP) kinase. Materials & Methods: IHOK was cultured three dimensionally with gingival fibroblast (GF), dermal fibroblast (DF) and immortalized gingival fibroblast (IGF). We characterized biologic properties of three dimensionally reconstructed IHOK by histological, immunohistochemical, and Western blot analysis. We also investigated whether MAP kinase pathway was involved in epithelial-mesenchymal interaction by Western blot analysis. Results: The best condition of three dimensionally cultured IHOK was the dermal equivalent consisting of type I collagen and IGF. IGF increased the expression of more proliferating cell nuclear antigen (PCNA), involucrin than GF and DF in response to co-culture with IHOK. Extracellularly regulated kinase (ERK) pathway was activated in organotypic co-culture with IGF. Conclusion: The organotypic co-culture of IHOK with dermal equivalent consisting of type I collagen and IGF resulted in excellent morphologic and immunohistochemical characteristics and involved ERK pathway. The epithelial-mesenchymal interaction was activated according to the fibroblast type.

Keywords

References

  1. Licciardello JT, Spitz MR, Hong WK: Multiple primary cancer in patients with cancer of the head and neck: Second cancer of the head and neck, esophagus and lung. Int J Radiat Oncol Biol Phys 1989;17:467-476 https://doi.org/10.1016/0360-3016(89)90096-5
  2. Oda D, Bigler L, Lee P, Blanton R: HPV immortalization of human oral cells: a model for carcinogenesis. Exp Cell Res 1996;226:164- 169 https://doi.org/10.1006/excr.1996.0215
  3. Yoo GH, Washington J, Piechocki M, Ensley J, Shibuya T, Oda D, et al.: The progression of head and neck cancer in vitro model. Arch Otolaryng Head Neck Surg 2000;126:1313-1318 https://doi.org/10.1001/archotol.126.11.1313
  4. Rheinwald JG, Green H: Serial cultivation of strains of human epidermal keratinocytes: the formation of keratinizing colonies from single cells. Cell 1975;6:331-343 https://doi.org/10.1016/S0092-8674(75)80001-8
  5. Levine JF, Stockdale EF: Cell-cell interactions promote mammary epithelial cell differentiation. J Cell Biol 1985;100:1415-1422 https://doi.org/10.1083/jcb.100.5.1415
  6. Boyce ST, Hansbrough JF: Biologic attachment, growth, and differentiation of cultured human epidermal keratinocytes on a graftable collagen and chondroitin-6 sulfate substrate. Surgery 1988;103:421-431
  7. Tiollier J, Dumas H, Tardy M, Tayot JL: Fibroblast behavior on gels of type I, III, and IV human placental collagens. Exp Cell Res 1990;191:95-104 https://doi.org/10.1016/0014-4827(90)90041-8
  8. Peault B: In-vitro models of stroma-dependent lymphopoiesis. Semi Immunol 1995;7:169-177 https://doi.org/10.1016/1044-5323(95)90044-6
  9. Hoffman RM: Three-dimensional histoculture: origins and applications in cancer reserch. Cancer Cell 1991;3:86-92
  10. Robbins KT, Connors KM, Storniolo AM, Hanchett C, Hoffman RM: Sponge-gel supported histoculture drug response assay for head and neck cancer. Correlations with clinical response to cisplatin. Arch Otolaryngol Head Neck Surg 1994;120:288-292 https://doi.org/10.1001/archotol.1994.01880270036007
  11. Robbins KT, Varki NM, Storniolo AM, Hoffman H, Hoffman RM: Drug response of head and neck tumors in native-state histoculture. Arch Otolaryngol Head Neck Surg 1991;117:83-86 https://doi.org/10.1001/archotol.1991.01870130089022
  12. Boukamp P, Breitkreutz D, Stark HJ, Fusenig NE: Mesenchymemediated and endogenous regulation of growth and differentiation of human skin keratinocytes derived from different body sites. Differentiation 1990;44:150-161 https://doi.org/10.1111/j.1432-0436.1990.tb00548.x
  13. Smola H, Thiekotter G, Fusenig NE: Mutual induction of growth factor gene expression by epidermal cell interaction. J Cell Biol 1993;122:417-429 https://doi.org/10.1083/jcb.122.2.417
  14. Schor SL, Schor AM: Clonal heterogeneity in fibroblast phenotype: implications for the control of epithelial-mesenchymal interactions. Bioessays 1987;7:200-204 https://doi.org/10.1002/bies.950070503
  15. Tajima S, Pinnell SR: Collagen synthesis by human skin fibroblasts in culture: studies of fibroblasts explanted from papillary and reticular dermis. J Invest Dermatol 1991;77:410-412
  16. Phipps RP, Borrello MA, Blieden TM: Fibroblast heterogeneity in the periodontium and other tissues. J Periodontal Res 1997;32:159-165 https://doi.org/10.1111/j.1600-0765.1997.tb01398.x
  17. Jiang WG, Hiscox S: Hepatocyte growth factor/scatter factor, a cytokine playing multiple and converse roles. Histopathol 1997; 12:537-555
  18. Finch PW, Rubin JS. Miki T, Ron D, Aaronson SA: Human KGF is FGF-related with properties of a paracrine effector of epithelial cell growth. Science 1989;245:752-755 https://doi.org/10.1126/science.2475908
  19. Dabelsteen S, Wandall HH, Gron B, Dabelsteen E: Keratinocyte growth factor mRNA expression in periodontal ligament fibroblasts. Eur J Oral Sci 1997;105:593-598 https://doi.org/10.1111/j.1600-0722.1997.tb00223.x
  20. Kim JM, Kim JS, Jung HC, Song IS, Kim CY: Up-regulation of inducible nitric oxide synthase and nitric oxide in Helicobacter pylori-infected human gastric epithelial cells:possible role of interferon- ${\gamma}$in polarized nitric oxide secretion. Helicobacter 2002;7:116-128 https://doi.org/10.1046/j.1083-4389.2002.00068.x
  21. Warny M, Keates A, Keates S, Castagliuolo I, Zacks JK, Aboudola S, et al: p38 MAP kinase activation by Clostridium difficile toxin A mediates monocyte necrosis, IL-8 production, and enteritis. J Clin Invest 2000;1147-1156 https://doi.org/10.1172/JCI7545
  22. Chiu LC, Kong CK, Ooi VE: The chlorophyllin-induced cell cycle arrest and apoptosis in human breast cancer MCF-7 cells is associated with ERK deactivation and Cyclin D1 depletion. Int J Mol Med. 2005;16:735-740
  23. Park NH, Min BM, Li SL, Huang MZ, Cherrick HM, Doniger J: Immortalization of normal human oral keratinocyte with type 16 human papillomavirus. Cariconogensis 1991;12:1627-1631 https://doi.org/10.1093/carcin/12.9.1627
  24. Dotto GP, Moellmann G, Ghosh S, Edwards M, Halaban RJ: Transformation of murine melanocytes by basic fibroblast growth factor cDNA and oncogenes and selective suppression of the transformed phenotype in a reconstituted cutaneous environment. J Cell Biol 1989;109:3115-3128 https://doi.org/10.1083/jcb.109.6.3115
  25. Vescio RA, Redfern CH, Nelson TJ. Ugoretz S, Stern PH, Hoffman RM: In vivo-like drug responses of human tumors growing in threedimensional, gel-supported, primary culture. Proc Natl Acad Sci USA 1987;84:5029-5033
  26. Stark HJ, Baur M, Breitkretz D, Mirancea N, Fusenig NE: Organotypic keratinocyte coculture I defined medium with regular epidermal morhogenesis and differentiation. J Invest Dermatol 1999;112:681-691 https://doi.org/10.1046/j.1523-1747.1999.00573.x
  27. Schoop VM, Mirancea N, Fusenig NE: Epidermal organization and differentiation of HaCaT keratinocytes in organotypic coculture with human dermal fibroblasts. J Inverst Dermatol 1999;112:343-353 https://doi.org/10.1046/j.1523-1747.1999.00524.x
  28. Demeter LM, Stoler MH, Broker TR, CHow LT: Induction of PCNA in differentiated kerationcytes of human papilloma virus infected lesions. Human Pathol 1994;25:343-348 https://doi.org/10.1016/0046-8177(94)90141-4
  29. Cheng S, Schmidt GD, Murant T, Broker TR, Chow LT: Differentiation dependent up regulation of the human papilloma virus E7 gene reactivates cellular DNA replication in suprabasilar differentiated keratinocytes. Genes Dev 1995;9:2335-2349 https://doi.org/10.1101/gad.9.19.2335
  30. Tavakkol A, Varani J, ELder JT, Zouboulis CC: Maintenace of human skin in organ culture: role for Insulin-like growth factor-1 receptor and EGFR. Arch Dermatol Res 1999;291:643-651 https://doi.org/10.1007/s004030050469
  31. Lee DY, Ahn HT, Cho KH: A new skin equivalent model: dermal substrate that combines de-epidermized dermis with fibroblasts-populated collagen matrix. J Dermatol Sci 2000;23:132-137 https://doi.org/10.1016/S0923-1811(00)00068-2
  32. Chung JH, Cho KH, Lee DY, Kwon OS, Sung MW, Kim KH, et al: Human oral buccal mucosa reconstructed on dermal substrates: a model for oral epithelial differentiation. Arch Dermatol Res 1997;289:677-685 https://doi.org/10.1007/s004030050261
  33. Hansson A, Bloor BK, Haig Y, Morgan PR, Etstrand J, Grafstrom RC: Expression of keratin in normal, immortalized and malignant oral epithelia in organotypic culture. Oral Oncol 2001;37:419-430 https://doi.org/10.1016/S1368-8375(00)00089-0
  34. Oda D, Savard CE, Eng L, Sekijima J, Haigh G, Lee SP: Reconstituted human oral and esophageal mucosa. In Vitro Cell Biol Anim 1998;34:46-52 https://doi.org/10.1007/s11626-998-0052-7
  35. Bloor BK, Su L, Shirlaw PJ, Morgan PR: Gene expression of the differentiation- specific keratins (4/13, and 1/10) in normal human buccal mucosa. Lab Invest 1998;78:787-795
  36. Su L, Lane EB, Morgan PR: Keratin 14 and 19 expression in normal, dysplastic and malignant oral epithelia. a study using in situ hybridization and immunohistochemistry. J Oral Pathol Med 1996;25:293-301 https://doi.org/10.1111/j.1600-0714.1996.tb00265.x
  37. Hietanen S, Syrajanen K, Syrajanen S: Chracterization of keratin and cell cycle protein expression in cell lines from squamous intraepithelial lesions progressing towards a malignant phenotype. Brit J cancer 1998;77:766-775 https://doi.org/10.1038/bjc.1998.125