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. 1996 Jan 1;313(Pt 1):201–206. doi: 10.1042/bj3130201

The role of cell adhesion in retinoic acid-induced modulation of chondrocyte phenotype.

M Sanchez 1, A Arcella 1, G Pontarelli 1, E Gionti 1
PMCID: PMC1216883  PMID: 8546684

Abstract

Retinoic acid (RA) treatment of a suspension of quail chondrocytes inhibits the expression of cartilage collagens and induces cell adhesion along with fibronectin expression. We asked whether the RA-induced modulation of the chondrocyte phenotype was dependent on cell adhesion. Prevention of cell adhesion blocks cell growth and many of the effects associated with RA, such as collagen II inhibition, collagen I activation and fibronectin induction. The activity of the bone/tendon promoter of the alpha 2(I) collagen gene was determined by measuring the transient expression of COL1A2-CAT, a chimaeric gene bearing 3500 bp from upstream of the transcription start site of the human alpha 2(I) gene fused to the chloramphenicol acetyltransferase (CAT) gene. This promoter is activated only in permissive conditions for cell adhesion. The attachment activities of chondrocytes on protein substrates was studied by an in vitro cell adhesion assay. Untreated cells or cells maintained in suspension while undergoing RA treatment do not attach when replated on protein substrates. Chondrocytes treated with RA in permissive conditions for cell adhesion rapidly attach and spread instead on collagen-coated wells. Altogether the results suggest that cell adhesion plays a major role in RA-induced modulation of the chondrocyte phenotype.

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Selected References

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  1. Ambesi-Impiombato F. S., Parks L. A., Coon H. G. Culture of hormone-dependent functional epithelial cells from rat thyroids. Proc Natl Acad Sci U S A. 1980 Jun;77(6):3455–3459. doi: 10.1073/pnas.77.6.3455. [DOI] [PMC free article] [PubMed] [Google Scholar]
  2. Beato M. Transcriptional control by nuclear receptors. FASEB J. 1991 Apr;5(7):2044–2051. doi: 10.1096/fasebj.5.7.2010057. [DOI] [PubMed] [Google Scholar]
  3. Bennett V. D., Adams S. L. Identification of a cartilage-specific promoter within intron 2 of the chick alpha 2(I) collagen gene. J Biol Chem. 1990 Feb 5;265(4):2223–2230. [PubMed] [Google Scholar]
  4. Benya P. D., Brown P. D., Padilla S. R. Microfilament modification by dihydrocytochalasin B causes retinoic acid-modulated chondrocytes to reexpress the differentiated collagen phenotype without a change in shape. J Cell Biol. 1988 Jan;106(1):161–170. doi: 10.1083/jcb.106.1.161. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Benya P. D., Padilla S. R. Modulation of the rabbit chondrocyte phenotype by retinoic acid terminates type II collagen synthesis without inducing type I collagen: the modulated phenotype differs from that produced by subculture. Dev Biol. 1986 Nov;118(1):296–305. doi: 10.1016/0012-1606(86)90096-5. [DOI] [PubMed] [Google Scholar]
  6. Boast S., Su M. W., Ramirez F., Sanchez M., Avvedimento E. V. Functional analysis of cis-acting DNA sequences controlling transcription of the human type I collagen genes. J Biol Chem. 1990 Aug 5;265(22):13351–13356. [PubMed] [Google Scholar]
  7. Brown P. D., Benya P. D. Alterations in chondrocyte cytoskeletal architecture during phenotypic modulation by retinoic acid and dihydrocytochalasin B-induced reexpression. J Cell Biol. 1988 Jan;106(1):171–179. doi: 10.1083/jcb.106.1.171. [DOI] [PMC free article] [PubMed] [Google Scholar]
  8. 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. doi: 10.1006/abio.1987.9999. [DOI] [PubMed] [Google Scholar]
  9. De Luca L. M. Retinoids and their receptors in differentiation, embryogenesis, and neoplasia. FASEB J. 1991 Nov;5(14):2924–2933. [PubMed] [Google Scholar]
  10. Dürr J., Goodman S., Potocnik A., von der Mark H., von der Mark K. Localization of beta 1-integrins in human cartilage and their role in chondrocyte adhesion to collagen and fibronectin. Exp Cell Res. 1993 Aug;207(2):235–244. doi: 10.1006/excr.1993.1189. [DOI] [PubMed] [Google Scholar]
  11. Enomoto M., Leboy P. S., Menko A. S., Boettiger D. Beta 1 integrins mediate chondrocyte interaction with type I collagen, type II collagen, and fibronectin. Exp Cell Res. 1993 Apr;205(2):276–285. doi: 10.1006/excr.1993.1087. [DOI] [PubMed] [Google Scholar]
  12. Fort P., Marty L., Piechaczyk M., el Sabrouty S., Dani C., Jeanteur P., Blanchard J. M. Various rat adult tissues express only one major mRNA species from the glyceraldehyde-3-phosphate-dehydrogenase multigenic family. Nucleic Acids Res. 1985 Mar 11;13(5):1431–1442. doi: 10.1093/nar/13.5.1431. [DOI] [PMC free article] [PubMed] [Google Scholar]
  13. Gerstenfeld L. C., Kelly C. M., Von Deck M., Lian J. B. Effect of 1,25-dihydroxyvitamin D3 on induction of chondrocyte maturation in culture: extracellular matrix gene expression and morphology. Endocrinology. 1990 Mar;126(3):1599–1609. doi: 10.1210/endo-126-3-1599. [DOI] [PubMed] [Google Scholar]
  14. Giguere V., Ong E. S., Segui P., Evans R. M. Identification of a receptor for the morphogen retinoic acid. Nature. 1987 Dec 17;330(6149):624–629. doi: 10.1038/330624a0. [DOI] [PubMed] [Google Scholar]
  15. Gionti E., Jullien P., Pontarelli G., Sanchez M. A continuous line of chicken embryo cells derived from a chondrocyte culture infected with RSV. Cell Differ Dev. 1989 Sep;27(3):215–223. doi: 10.1016/0922-3371(89)90701-6. [DOI] [PubMed] [Google Scholar]
  16. 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. doi: 10.1073/pnas.82.9.2756. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Gorman C. M., Merlino G. T., Willingham M. C., Pastan I., Howard B. H. The Rous sarcoma virus long terminal repeat is a strong promoter when introduced into a variety of eukaryotic cells by DNA-mediated transfection. Proc Natl Acad Sci U S A. 1982 Nov;79(22):6777–6781. doi: 10.1073/pnas.79.22.6777. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Gorman C. M., Moffat L. F., Howard B. H. Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol. 1982 Sep;2(9):1044–1051. doi: 10.1128/mcb.2.9.1044. [DOI] [PMC free article] [PubMed] [Google Scholar]
  19. Gudas L. J. Retinoids and vertebrate development. J Biol Chem. 1994 Jun 3;269(22):15399–15402. [PubMed] [Google Scholar]
  20. Gudas L. J. Retinoids, retinoid-responsive genes, cell differentiation, and cancer. Cell Growth Differ. 1992 Sep;3(9):655–662. [PubMed] [Google Scholar]
  21. Horton W. E., Yamada Y., Hassell J. R. Retinoic acid rapidly reduces cartilage matrix synthesis by altering gene transcription in chondrocytes. Dev Biol. 1987 Oct;123(2):508–516. doi: 10.1016/0012-1606(87)90409-x. [DOI] [PubMed] [Google Scholar]
  22. Horton W., Hassell J. R. Independence of cell shape and loss of cartilage matrix production during retinoic acid treatment of cultured chondrocytes. Dev Biol. 1986 Jun;115(2):392–397. doi: 10.1016/0012-1606(86)90258-7. [DOI] [PubMed] [Google Scholar]
  23. Iwamoto M., Golden E. B., Adams S. L., Noji S., Pacifici M. Responsiveness to retinoic acid changes during chondrocyte maturation. Exp Cell Res. 1993 Apr;205(2):213–224. doi: 10.1006/excr.1993.1079. [DOI] [PubMed] [Google Scholar]
  24. Kliewer S. A., Umesono K., Mangelsdorf D. J., Evans R. M. Retinoid X receptor interacts with nuclear receptors in retinoic acid, thyroid hormone and vitamin D3 signalling. Nature. 1992 Jan 30;355(6359):446–449. doi: 10.1038/355446a0. [DOI] [PMC free article] [PubMed] [Google Scholar]
  25. Lehrach H., Diamond D., Wozney J. M., Boedtker H. RNA molecular weight determinations by gel electrophoresis under denaturing conditions, a critical reexamination. Biochemistry. 1977 Oct 18;16(21):4743–4751. doi: 10.1021/bi00640a033. [DOI] [PubMed] [Google Scholar]
  26. Mallein-Gerin F., Garrone R., van der Rest M. Proteoglycan and collagen synthesis are correlated with actin organization in dedifferentiating chondrocytes. Eur J Cell Biol. 1991 Dec;56(2):364–373. [PubMed] [Google Scholar]
  27. Marchisio P. C., Capasso O., Nitsch L., Cancedda R., Gionti E. Cytoskeleton and adhesion patterns of cultured chick embryo chondrocytes during cell spreading and Rous sarcoma virus transformation. Exp Cell Res. 1984 Apr;151(2):332–343. doi: 10.1016/0014-4827(84)90384-7. [DOI] [PubMed] [Google Scholar]
  28. Mayne R., Elrod B. W., Mayne P. M., Sanderson R. D., Linsenmayer T. F. Changes in the synthesis of minor cartilage collagens after growth of chick chondrocytes in 5-bromo-2'-deoxyuridine or to senescence. Exp Cell Res. 1984 Mar;151(1):171–182. doi: 10.1016/0014-4827(84)90366-5. [DOI] [PubMed] [Google Scholar]
  29. Mourey M. S., Quadro L., Panariello L., Colantuoni V. Retinoids regulate expression of the retinol-binding protein gene in hepatoma cells in culture. J Cell Physiol. 1994 Sep;160(3):596–602. doi: 10.1002/jcp.1041600323. [DOI] [PubMed] [Google Scholar]
  30. Ninomiya Y., Olsen B. R. Synthesis and characterization of cDNA encoding a cartilage-specific short collagen. Proc Natl Acad Sci U S A. 1984 May;81(10):3014–3018. doi: 10.1073/pnas.81.10.3014. [DOI] [PMC free article] [PubMed] [Google Scholar]
  31. Norton P. A., Hynes R. O. Alternative splicing of chicken fibronectin in embryos and in normal and transformed cells. Mol Cell Biol. 1987 Dec;7(12):4297–4307. doi: 10.1128/mcb.7.12.4297. [DOI] [PMC free article] [PubMed] [Google Scholar]
  32. Oettinger H. F., Pacifici M. Type X collagen gene expression is transiently up-regulated by retinoic acid treatment in chick chondrocyte cultures. Exp Cell Res. 1990 Dec;191(2):292–298. doi: 10.1016/0014-4827(90)90017-5. [DOI] [PubMed] [Google Scholar]
  33. Petkovich M., Brand N. J., Krust A., Chambon P. A human retinoic acid receptor which belongs to the family of nuclear receptors. Nature. 1987 Dec 3;330(6147):444–450. doi: 10.1038/330444a0. [DOI] [PubMed] [Google Scholar]
  34. Ross S. A., Ahrens R. A., De Luca L. M. Retinoic acid enhances adhesiveness, laminin and integrin beta 1 synthesis, and retinoic acid receptor expression in F9 teratocarcinoma cells. J Cell Physiol. 1994 May;159(2):263–273. doi: 10.1002/jcp.1041590210. [DOI] [PubMed] [Google Scholar]
  35. Sanchez M., Gionti E., Arcella A., Pontarelli G., De Lorenzo F. Alpha 2(I) collagen gene expression is up-regulated in quail chondrocytes pretreated with retinoic acid. Biochem J. 1993 Oct 1;295(Pt 1):115–119. doi: 10.1042/bj2950115. [DOI] [PMC free article] [PubMed] [Google Scholar]
  36. Sanchez M., Gionti E., Pontarelli G., Arcella A., De Lorenzo F. Expression of type X collagen is transiently stimulated in redifferentiating chondrocytes pretreated with retinoic acid. Biochem J. 1991 May 15;276(Pt 1):183–187. doi: 10.1042/bj2760183. [DOI] [PMC free article] [PubMed] [Google Scholar]
  37. Tamkun J. W., DeSimone D. W., Fonda D., Patel R. S., Buck C., Horwitz A. F., Hynes R. O. Structure of integrin, a glycoprotein involved in the transmembrane linkage between fibronectin and actin. Cell. 1986 Jul 18;46(2):271–282. doi: 10.1016/0092-8674(86)90744-0. [DOI] [PubMed] [Google Scholar]
  38. Tuckwell D. S., Ayad S., Grant M. E., Takigawa M., Humphries M. J. Conformation dependence of integrin-type II collagen binding. Inability of collagen peptides to support alpha 2 beta 1 binding, and mediation of adhesion to denatured collagen by a novel alpha 5 beta 1-fibronectin bridge. J Cell Sci. 1994 Apr;107(Pt 4):993–1005. doi: 10.1242/jcs.107.4.993. [DOI] [PubMed] [Google Scholar]
  39. Vertel B. M., Dorfman A. Simultaneous localization of type II collagen and core protein of chondroitin sulfate proteoglycan in individual chondrocytes. Proc Natl Acad Sci U S A. 1979 Mar;76(3):1261–1264. doi: 10.1073/pnas.76.3.1261. [DOI] [PMC free article] [PubMed] [Google Scholar]
  40. Yasui N., Benya P. D., Nimni M. E. Coordinate regulation of type IX and type II collagen synthesis during growth of chick chondrocytes in retinoic acid or 5-bromo-2'-deoxyuridine. J Biol Chem. 1986 Jun 15;261(17):7997–8001. [PubMed] [Google Scholar]
  41. Yu V. C., Delsert C., Andersen B., Holloway J. M., Devary O. V., När A. M., Kim S. Y., Boutin J. M., Glass C. K., Rosenfeld M. G. RXR beta: a coregulator that enhances binding of retinoic acid, thyroid hormone, and vitamin D receptors to their cognate response elements. Cell. 1991 Dec 20;67(6):1251–1266. doi: 10.1016/0092-8674(91)90301-e. [DOI] [PubMed] [Google Scholar]
  42. Zhang X. K., Hoffmann B., Tran P. B., Graupner G., Pfahl M. Retinoid X receptor is an auxiliary protein for thyroid hormone and retinoic acid receptors. Nature. 1992 Jan 30;355(6359):441–446. doi: 10.1038/355441a0. [DOI] [PubMed] [Google Scholar]
  43. de The H., Marchio A., Tiollais P., Dejean A. Differential expression and ligand regulation of the retinoic acid receptor alpha and beta genes. EMBO J. 1989 Feb;8(2):429–433. doi: 10.1002/j.1460-2075.1989.tb03394.x. [DOI] [PMC free article] [PubMed] [Google Scholar]
  44. de Thé H., Vivanco-Ruiz M. M., Tiollais P., Stunnenberg H., Dejean A. Identification of a retinoic acid responsive element in the retinoic acid receptor beta gene. Nature. 1990 Jan 11;343(6254):177–180. doi: 10.1038/343177a0. [DOI] [PubMed] [Google Scholar]
  45. von der Mark K., Gauss V., von der Mark H., Müller P. Relationship between cell shape and type of collagen synthesised as chondrocytes lose their cartilage phenotype in culture. Nature. 1977 Jun 9;267(5611):531–532. doi: 10.1038/267531a0. [DOI] [PubMed] [Google Scholar]

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