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J Clin Invest. 1994 Dec; 94(6): 2307–2316.
PMCID: PMC330059

Interleukin-1 beta-modulated gene expression in immortalized human chondrocytes.


Immortalized human chondrocytes were established by transfection of primary cultures of juvenile costal chondrocytes with vectors encoding simian virus 40 large T antigen and selection in suspension culture over agarose. Stable cell lines were generated that exhibited chondrocyte morphology, continuous proliferative capacity (> 80 passages) in monolayer culture in serum-containing medium, and expression of mRNAs encoding chondrocyte-specific collagens II, IX, and XI and proteoglycans in an insulin-containing serum substitute. They did not express type X collagen or versican mRNA. These cells synthesized and secreted extracellular matrix molecules that were reactive with monoclonal antibodies against type II collagen, large proteoglycan (PG-H, aggrecan), and chondroitin-4- and chondroitin-6-sulfate. Interleukin-1 beta (IL-1 beta) decreased the levels of type II collagen mRNA and increased the levels of mRNAs for collagenase, stromelysin, and immediate early genes (egr-1, c-fos, c-jun, and jun-B). These cell lines also expressed reporter gene constructs containing regulatory sequences (-577/+3,428 bp) of the type II collagen gene (COL2A1) in transient transfection experiments, and IL-1 beta suppressed this expression by 50-80%. These results show that immortalized human chondrocytes displaying cartilage-specific modulation by IL-1 beta can be used as a model for studying normal and pathological repair mechanisms.

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  • Goldring MB, Sandell LJ, Stephenson ML, Krane SM. Immune interferon suppresses levels of procollagen mRNA and type II collagen synthesis in cultured human articular and costal chondrocytes. J Biol Chem. 1986 Jul 5;261(19):9049–9055. [PubMed]
  • Goldring MB, Birkhead J, Sandell LJ, Kimura T, Krane SM. Interleukin 1 suppresses expression of cartilage-specific types II and IX collagens and increases types I and III collagens in human chondrocytes. J Clin Invest. 1988 Dec;82(6):2026–2037. [PMC free article] [PubMed]
  • Goldring MB, Krane SM. Modulation by recombinant interleukin 1 of synthesis of types I and III collagens and associated procollagen mRNA levels in cultured human cells. J Biol Chem. 1987 Dec 5;262(34):16724–16729. [PubMed]
  • Benya PD, Shaffer JD. Dedifferentiated chondrocytes reexpress the differentiated collagen phenotype when cultured in agarose gels. Cell. 1982 Aug;30(1):215–224. [PubMed]
  • Adolphe M, Froger B, Ronot X, Corvol MT, Forest N. Cell multiplication and type II collagen production by rabbit articular chondrocytes cultivated in a defined medium. Exp Cell Res. 1984 Dec;155(2):527–536. [PubMed]
  • Castagnola P, Moro G, Descalzi-Cancedda F, Cancedda R. Type X collagen synthesis during in vitro development of chick embryo tibial chondrocytes. J Cell Biol. 1986 Jun;102(6):2310–2317. [PMC free article] [PubMed]
  • Alema S, Tato F, Boettiger D. myc and src oncogenes have complementary effects on cell proliferation and expression of specific extracellular matrix components in definitive chondroblasts. Mol Cell Biol. 1985 Mar;5(3):538–544. [PMC free article] [PubMed]
  • Gionti E, Pontarelli G, Cancedda R. Avian myelocytomatosis virus immortalizes differentiated quail chondrocytes. Proc Natl Acad Sci U S A. 1985 May;82(9):2756–2760. [PMC free article] [PubMed]
  • Horton WE, Jr, Cleveland J, Rapp U, Nemuth G, Bolander M, Doege K, Yamada Y, Hassell JR. An established rat cell line expressing chondrocyte properties. Exp Cell Res. 1988 Oct;178(2):457–468. [PubMed]
  • Thenet S, Benya PD, Demignot S, Feunteun J, Adolphe M. SV40-immortalization of rabbit articular chondrocytes: alteration of differentiated functions. J Cell Physiol. 1992 Jan;150(1):158–167. [PubMed]
  • Takigawa M, Tajima K, Pan HO, Enomoto M, Kinoshita A, Suzuki F, Takano Y, Mori Y. Establishment of a clonal human chondrosarcoma cell line with cartilage phenotypes. Cancer Res. 1989 Jul 15;49(14):3996–4002. [PubMed]
  • Block JA, Inerot SE, Gitelis S, Kimura JH. Synthesis of chondrocytic keratan sulphate-containing proteoglycans by human chondrosarcoma cells in long-term cell culture. J Bone Joint Surg Am. 1991 Jun;73(5):647–658. [PubMed]
  • Grigoriadis AE, Heersche JN, Aubin JE. Differentiation of muscle, fat, cartilage, and bone from progenitor cells present in a bone-derived clonal cell population: effect of dexamethasone. J Cell Biol. 1988 Jun;106(6):2139–2151. [PMC free article] [PubMed]
  • Bernier SM, Goltzman D. Regulation of expression of the chondrocytic phenotype in a skeletal cell line (CFK2) in vitro. J Bone Miner Res. 1993 Apr;8(4):475–484. [PubMed]
  • Gerstenfeld LC, Kelly CM, Von Deck M, Lian JB. Comparative morphological and biochemical analysis of hypertrophic, non-hypertrophic and 1,25(OH)2D3 treated non-hypertrophic chondrocytes. Connect Tissue Res. 1990;24(1):29–39. [PubMed]
  • Adams SL, Pallante KM, Niu Z, Leboy PS, Golden EB, Pacifici M. Rapid induction of type X collagen gene expression in cultured chick vertebral chondrocytes. Exp Cell Res. 1991 Mar;193(1):190–197. [PubMed]
  • Tyler JA, Benton HP. Synthesis of type II collagen is decreased in cartilage cultured with interleukin 1 while the rate of intracellular degradation remains unchanged. Coll Relat Res. 1988 Sep;8(5):393–405. [PubMed]
  • Yaron I, Meyer FA, Dayer JM, Bleiberg I, Yaron M. Some recombinant human cytokines stimulate glycosaminoglycan synthesis in human synovial fibroblast cultures and inhibit it in human articular cartilage cultures. Arthritis Rheum. 1989 Feb;32(2):173–180. [PubMed]
  • Tyler JA. Articular cartilage cultured with catabolin (pig interleukin 1) synthesizes a decreased number of normal proteoglycan molecules. Biochem J. 1985 May 1;227(3):869–878. [PMC free article] [PubMed]
  • Ikebe T, Hirata M, Koga T. Effects of human recombinant tumor necrosis factor-alpha and interleukin 1 on the synthesis of glycosaminoglycan and DNA in cultured rat costal chondrocytes. J Immunol. 1988 Feb 1;140(3):827–831. [PubMed]
  • Kimura T, Mattei MG, Stevens JW, Goldring MB, Ninomiya Y, Olsen BR. Molecular cloning of rat and human type IX collagen cDNA and localization of the alpha 1(IX) gene on the human chromosome 6. Eur J Biochem. 1989 Jan 15;179(1):71–78. [PubMed]
  • Kimura T, Cheah KS, Chan SD, Lui VC, Mattei MG, van der Rest M, Ono K, Solomon E, Ninomiya Y, Olsen BR. The human alpha 2(XI) collagen (COL11A2) chain. Molecular cloning of cDNA and genomic DNA reveals characteristics of a fibrillar collagen with differences in genomic organization. J Biol Chem. 1989 Aug 15;264(23):13910–13916. [PubMed]
  • Chu ML, Myers JC, Bernard MP, Ding JF, Ramirez F. Cloning and characterization of five overlapping cDNAs specific for the human pro alpha 1(I) collagen chain. Nucleic Acids Res. 1982 Oct 11;10(19):5925–5934. [PMC free article] [PubMed]
  • Doege KJ, Sasaki M, Kimura T, Yamada Y. Complete coding sequence and deduced primary structure of the human cartilage large aggregating proteoglycan, aggrecan. Human-specific repeats, and additional alternatively spliced forms. J Biol Chem. 1991 Jan 15;266(2):894–902. [PubMed]
  • Day AA, Ramis CI, Fisher LW, Gehron-Robey P, Termine JD, Young MF. Characterization of bone PG II cDNA and its relationship to PG II mRNA from other connective tissues. Nucleic Acids Res. 1986 Dec 22;14(24):9861–9876. [PMC free article] [PubMed]
  • Fisher LW, Heegaard AM, Vetter U, Vogel W, Just W, Termine JD, Young MF. Human biglycan gene. Putative promoter, intron-exon junctions, and chromosomal localization. J Biol Chem. 1991 Aug 5;266(22):14371–14377. [PubMed]
  • Stephenson ML, Goldring MB, Birkhead JR, Krane SM, Rahmsdorf HJ, Angel P. Stimulation of procollagenase synthesis parallels increases in cellular procollagenase mRNA in human articular chondrocytes exposed to recombinant interleukin 1 beta or phorbol ester. Biochem Biophys Res Commun. 1987 Apr 29;144(2):583–590. [PubMed]
  • MacNaul KL, Chartrain N, Lark M, Tocci MJ, Hutchinson NI. Discoordinate expression of stromelysin, collagenase, and tissue inhibitor of metalloproteinases-1 in rheumatoid human synovial fibroblasts. Synergistic effects of interleukin-1 and tumor necrosis factor-alpha on stromelysin expression. J Biol Chem. 1990 Oct 5;265(28):17238–17245. [PubMed]
  • Nakabeppu Y, Ryder K, Nathans D. DNA binding activities of three murine Jun proteins: stimulation by Fos. Cell. 1988 Dec 2;55(5):907–915. [PubMed]
  • Ryder K, Lau LF, Nathans D. A gene activated by growth factors is related to the oncogene v-jun. Proc Natl Acad Sci U S A. 1988 Mar;85(5):1487–1491. [PMC free article] [PubMed]
  • Curran T, MacConnell WP, van Straaten F, Verma IM. Structure of the FBJ murine osteosarcoma virus genome: molecular cloning of its associated helper virus and the cellular homolog of the v-fos gene from mouse and human cells. Mol Cell Biol. 1983 May;3(5):914–921. [PMC free article] [PubMed]
  • Sukhatme VP, Cao XM, Chang LC, Tsai-Morris CH, Stamenkovich D, Ferreira PC, Cohen DR, Edwards SA, Shows TB, Curran T, et al. A zinc finger-encoding gene coregulated with c-fos during growth and differentiation, and after cellular depolarization. Cell. 1988 Apr 8;53(1):37–43. [PubMed]
  • Small MB, Gluzman Y, Ozer HL. Enhanced transformation of human fibroblasts by origin-defective simian virus 40. Nature. 1982 Apr 15;296(5858):671–672. [PubMed]
  • Chaney WG, Howard DR, Pollard JW, Sallustio S, Stanley P. High-frequency transfection of CHO cells using polybrene. Somat Cell Mol Genet. 1986 May;12(3):237–244. [PubMed]
  • Arbiser JL, Arbiser ZK, Majzoub JA. Regulation of gene expression in choriocarcinoma by methotrexate and hydroxyurea. Endocrinology. 1991 Feb;128(2):972–978. [PubMed]
  • Williams DA, Rosenblatt MF, Beier DR, Cone RD. Generation of murine stromal cell lines supporting hematopoietic stem cell proliferation by use of recombinant retrovirus vectors encoding simian virus 40 large T antigen. Mol Cell Biol. 1988 Sep;8(9):3864–3871. [PMC free article] [PubMed]
  • Apperley JF, Luskey BD, Williams DA. Retroviral gene transfer of human adenosine deaminase in murine hematopoietic cells: effect of selectable marker sequences on long-term expression. Blood. 1991 Jul 15;78(2):310–317. [PubMed]
  • Asari A, Miyauchi S, Miyazaki K, Hamai A, Horie K, Takahashi T, Sekiguchi T, Machida A, Kohno K, Uchiyama Y. Intra- and extracellular localization of hyaluronic acid and proteoglycan constituents (chondroitin sulfate, keratan sulfate, and protein core) in articular cartilage of rabbit tibia. J Histochem Cytochem. 1992 Nov;40(11):1693–1704. [PubMed]
  • Chomczynski P, Sacchi N. Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem. 1987 Apr;162(1):156–159. [PubMed]
  • Cathala G, Savouret JF, Mendez B, West BL, Karin M, Martial JA, Baxter JD. A method for isolation of intact, translationally active ribonucleic acid. DNA. 1983;2(4):329–335. [PubMed]
  • Goldring MB, Fukuo K, Birkhead JR, Dudek E, Sandell LJ. Transcriptional suppression by interleukin-1 and interferon-gamma of type II collagen gene expression in human chondrocytes. J Cell Biochem. 1994 Jan;54(1):85–99. [PubMed]
  • Ryan MC, Sieraski M, Sandell LJ. The human type II procollagen gene: identification of an additional protein-coding domain and location of potential regulatory sequences in the promoter and first intron. Genomics. 1990 Sep;8(1):41–48. [PubMed]
  • Murnane JP, Fuller LF, Painter RB. Establishment and characterization of a permanent pSV ori--transformed ataxia-telangiectasia cell line. Exp Cell Res. 1985 May;158(1):119–126. [PubMed]
  • Mallein-Gerin F, Olsen BR. Expression of simian virus 40 large T (tumor) oncogene in mouse chondrocytes induces cell proliferation without loss of the differentiated phenotype. Proc Natl Acad Sci U S A. 1993 Apr 15;90(8):3289–3293. [PMC free article] [PubMed]
  • Takigawa M, Pan HO, Kinoshita A, Tajima K, Takano Y. Establishment from a human chondrosarcoma of a new immortal cell line with high tumorigenicity in vivo, which is able to form proteoglycan-rich cartilage-like nodules and to respond to insulin in vitro. Int J Cancer. 1991 Jul 9;48(5):717–725. [PubMed]
  • Cizdziel PE, Hosoi J, Montgomery JC, Wiseman RW, Barrett JC. Loss of a tumor suppressor gene function is correlated with downregulation of chondrocyte-specific collagen expression in Syrian hamster embryo cells. Mol Carcinog. 1991;4(1):14–24. [PubMed]
  • Kato Y, Nasu N, Takase T, Daikuhara Y, Suzuki F. A serum-free medium supplemented with multiplication-stimulating activity (MSA) supports both proliferation and differentiation of chondrocytes in primary culture. Exp Cell Res. 1980 Jan;125(1):167–174. [PubMed]
  • Stevens RL, Nissley SP, Kimura JH, Rechler MM, Caplan AI, Hascall VC. Effects of insulin and multiplication-stimulating activity on proteoglycan biosynthesis in chondrocytes from the Swarm rat chondrosarcoma. J Biol Chem. 1981 Feb 25;256(4):2045–2052. [PubMed]
  • Böhme K, Conscience-Egli M, Tschan T, Winterhalter KH, Bruckner P. Induction of proliferation or hypertrophy of chondrocytes in serum-free culture: the role of insulin-like growth factor-I, insulin, or thyroxine. J Cell Biol. 1992 Feb;116(4):1035–1042. [PMC free article] [PubMed]
  • Glaser JH, Conrad HE. Properties of chick embryo chondrocytes grown in serum-free medium. J Biol Chem. 1984 Jun 10;259(11):6766–6772. [PubMed]
  • Hale LV, Hale JE, Kemick ML, Ishikawa Y, Wuthier RE. Development of a new serum-free medium, USC-HC1, for growth and normal phenotype in postembryonic chicken growth plate chondrocytes. In Vitro Cell Dev Biol. 1986 Oct;22(10):597–603. [PubMed]
  • Paulsen DF, Solursh M. Microtiter micromass cultures of limb-bud mesenchymal cells. In Vitro Cell Dev Biol. 1988 Feb;24(2):138–147. [PubMed]
  • Carrino DA, Kujawa MJ, Lennon DP, Caplan AI. Altered cartilage proteoglycans synthesized by chick limb bud chondrocytes cultured in serum-free defined medium. Exp Cell Res. 1989 Jul;183(1):62–71. [PubMed]
  • Rosselot G, Reginato AM, Leach RM. Development of a serum-free system to study the effect of growth hormone and insulinlike growth factor-I on cultured postembryonic growth plate chondrocytes. In Vitro Cell Dev Biol. 1992 Apr;28A(4):235–244. [PubMed]
  • Niesor EJ, Wollheim CB, Mintz DH, Blondel B, Renold AE, Weil R. Establishment of rat pancreatic endocrine cell lines by infection with simian virus 40. Biochem J. 1979 Mar 15;178(3):559–568. [PMC free article] [PubMed]
  • Trojan J, Blossey BK, Johnson TR, Rudin SD, Tykocinski M, Ilan J, Ilan J. Loss of tumorigenicity of rat glioblastoma directed by episome-based antisense cDNA transcription of insulin-like growth factor I. Proc Natl Acad Sci U S A. 1992 Jun 1;89(11):4874–4878. [PMC free article] [PubMed]
  • Medema RH, Wubbolts R, Bos JL. Two dominant inhibitory mutants of p21ras interfere with insulin-induced gene expression. Mol Cell Biol. 1991 Dec;11(12):5963–5967. [PMC free article] [PubMed]
  • Sandell LJ, Morris N, Robbins JR, Goldring MB. Alternatively spliced type II procollagen mRNAs define distinct populations of cells during vertebral development: differential expression of the amino-propeptide. J Cell Biol. 1991 Sep;114(6):1307–1319. [PMC free article] [PubMed]
  • McMahon AP, Champion JE, McMahon JA, Sukhatme VP. Developmental expression of the putative transcription factor Egr-1 suggests that Egr-1 and c-fos are coregulated in some tissues. Development. 1990 Feb;108(2):281–287. [PubMed]
  • Caubet JF, Bernaudin JF. Expression of the c-fos proto-oncogene in bone, cartilage and tooth forming tissues during mouse development. Biol Cell. 1988;64(1):101–104. [PubMed]
  • Wilkinson DG, Bhatt S, Ryseck RP, Bravo R. Tissue-specific expression of c-jun and junB during organogenesis in the mouse. Development. 1989 Jul;106(3):465–471. [PubMed]
  • Kato Y, Nakashima K, Iwamoto M, Murakami H, Hiranuma H, Koike T, Suzuki F, Fuchihata H, Ikehara Y, Noshiro M, et al. Effects of interleukin-1 on syntheses of alkaline phosphatase, type X collagen, and 1,25-dihydroxyvitamin D3 receptor, and matrix calcification in rabbit chondrocyte cultures. J Clin Invest. 1993 Nov;92(5):2323–2330. [PMC free article] [PubMed]

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