Skip to main content
NCBI home page
Biochemical Journal logoLink to Biochemical Journal
. 2001 Feb 15;354(Pt 1):17–24. doi: 10.1042/0264-6021:3540017

Differential regulation and expression of hyaluronan synthases in human articular chondrocytes, synovial cells and osteosarcoma cells.

A D Recklies 1, C White 1, L Melching 1, P J Roughley 1
PMCID: PMC1221623  PMID: 11171074

Abstract

Recently three isoforms of hyaluronan synthase (HAS), the enzyme responsible for hyaluronate/hyaluronan (HA) biosynthesis, have been cloned, allowing us to study their expression pattern. Our objective was to determine which of the HAS isoenzymes were expressed in human articular chondrocytes, synovial fibroblasts and osteosarcoma cells, whether their expression could be modulated by growth factors (insulin-like growth factor-1, basic fibroblast growth factor and transforming growth factor (TGF-beta1) and cytokines [interleukin 1beta1 (IL-1beta)], and whether changes in the rate of HA synthesis by the cells correlated with changes in mRNA levels for one or more of the HAS isoforms. All three HAS isoforms were found to be expressed in the cultured cells analysed in this study, although the relative proportions varied for each cell type. HAS2 mRNA was usually predominant in chondrocytes, whereas synovial cells contained increased amounts of HAS1. HAS3 was always the least abundant message. The rapidly growing osteosarcoma cells contained almost exclusively HAS2 message. HAS usage in uncultured cartilage and synovial tissues was similar to that in the cultured cells, with HAS2 message being the predominant species in cartilage and HAS1 usually being the predominant species in synovium. HA synthesis was stimulated by the growth factors, but the extent of the response was cell-type specific. Synovial cells responded particularly well to IL-1beta, and showed a unique synergistic response when IL-1beta was used in combination with TGF-beta1. This response was much reduced in articular chondrocytes and absent in the osteosarcoma cells. Analysis of changes in HAS message levels indicated that there was often no correlation with the changes in HA secretion following exposure to growth factors. Although HAS-1 mRNA was increased in synovial cells after exposure to TGF-beta1/IL-1beta, the magnitude of the change was far less than the effect on HA synthesis. Our data thus suggest that HAS gene usage is tissue specific, and the regulation by growth factors is unique for each HAS gene and is further modulated by cell-specific factors. In addition, regulation of HA biosynthesis appears to be multi-faceted, with control of HAS gene expression and mRNA levels being only one aspect of this process.

Full Text

The Full Text of this article is available as a PDF (147.4 KB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Asplund T., Brinck J., Suzuki M., Briskin M. J., Heldin P. Characterization of hyaluronan synthase from a human glioma cell line. Biochim Biophys Acta. 1998 May 8;1380(3):377–388. doi: 10.1016/s0304-4165(98)00010-5. [DOI] [PubMed] [Google Scholar]
  2. Butler D. M., Vitti G. F., Leizer T., Hamilton J. A. Stimulation of the hyaluronic acid levels of human synovial fibroblasts by recombinant human tumor necrosis factor alpha, tumor necrosis factor beta (lymphotoxin), interleukin-1 alpha, and interleukin-1 beta. Arthritis Rheum. 1988 Oct;31(10):1281–1289. doi: 10.1002/art.1780311010. [DOI] [PubMed] [Google Scholar]
  3. 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]
  4. Clark R. A., Alon R., Springer T. A. CD44 and hyaluronan-dependent rolling interactions of lymphocytes on tonsillar stroma. J Cell Biol. 1996 Aug;134(4):1075–1087. doi: 10.1083/jcb.134.4.1075. [DOI] [PMC free article] [PubMed] [Google Scholar]
  5. Clarris B. J., Fraser J. R. On the pericellular zone of some mammalian cells in vitro. Exp Cell Res. 1968 Jan;49(1):181–193. doi: 10.1016/0014-4827(68)90530-2. [DOI] [PubMed] [Google Scholar]
  6. DeGrendele H. C., Estess P., Picker L. J., Siegelman M. H. CD44 and its ligand hyaluronate mediate rolling under physiologic flow: a novel lymphocyte-endothelial cell primary adhesion pathway. J Exp Med. 1996 Mar 1;183(3):1119–1130. doi: 10.1084/jem.183.3.1119. [DOI] [PMC free article] [PubMed] [Google Scholar]
  7. Dougherty B. A., van de Rijn I. Molecular characterization of hasA from an operon required for hyaluronic acid synthesis in group A streptococci. J Biol Chem. 1994 Jan 7;269(1):169–175. [PubMed] [Google Scholar]
  8. Edwards J. C., Wilkinson L. S., Jones H. M., Soothill P., Henderson K. J., Worrall J. G., Pitsillides A. A. The formation of human synovial joint cavities: a possible role for hyaluronan and CD44 in altered interzone cohesion. J Anat. 1994 Oct;185(Pt 2):355–367. [PMC free article] [PubMed] [Google Scholar]
  9. Goldberg R. L. Enzyme-linked immunosorbent assay for hyaluronate using cartilage proteoglycan and an antibody to keratan sulfate. Anal Biochem. 1988 Nov 1;174(2):448–458. doi: 10.1016/0003-2697(88)90043-7. [DOI] [PubMed] [Google Scholar]
  10. Golds E. E., Mason P., Nyirkos P. Inflammatory cytokines induce synthesis and secretion of gro protein and a neutrophil chemotactic factor but not beta 2-microglobulin in human synovial cells and fibroblasts. Biochem J. 1989 Apr 15;259(2):585–588. doi: 10.1042/bj2590585. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Hiscock D. R., Caterson B., Flannery C. R. Expression of hyaluronan synthases in articular cartilage. Osteoarthritis Cartilage. 2000 Mar;8(2):120–126. doi: 10.1053/joca.1999.0280. [DOI] [PubMed] [Google Scholar]
  12. Itano N., Kimata K. Expression cloning and molecular characterization of HAS protein, a eukaryotic hyaluronan synthase. J Biol Chem. 1996 Apr 26;271(17):9875–9878. doi: 10.1074/jbc.271.17.9875. [DOI] [PubMed] [Google Scholar]
  13. Itano N., Kimata K. Molecular cloning of human hyaluronan synthase. Biochem Biophys Res Commun. 1996 May 24;222(3):816–820. doi: 10.1006/bbrc.1996.0827. [DOI] [PubMed] [Google Scholar]
  14. Itano N., Sawai T., Yoshida M., Lenas P., Yamada Y., Imagawa M., Shinomura T., Hamaguchi M., Yoshida Y., Ohnuki Y. Three isoforms of mammalian hyaluronan synthases have distinct enzymatic properties. J Biol Chem. 1999 Aug 27;274(35):25085–25092. doi: 10.1074/jbc.274.35.25085. [DOI] [PubMed] [Google Scholar]
  15. Knudson C. B., Knudson W. Hyaluronan-binding proteins in development, tissue homeostasis, and disease. FASEB J. 1993 Oct;7(13):1233–1241. [PubMed] [Google Scholar]
  16. Knudson W., Bartnik E., Knudson C. B. Assembly of pericellular matrices by COS-7 cells transfected with CD44 lymphocyte-homing receptor genes. Proc Natl Acad Sci U S A. 1993 May 1;90(9):4003–4007. doi: 10.1073/pnas.90.9.4003. [DOI] [PMC free article] [PubMed] [Google Scholar]
  17. Melching L. I., Cs-Szabo G., Roughley P. J. Analysis of proteoglycan messages in human articular cartilage by a competitive PCR technique. Matrix Biol. 1997 Apr;16(1):1–11. doi: 10.1016/s0945-053x(97)90111-6. [DOI] [PubMed] [Google Scholar]
  18. Mikecz K., Brennan F. R., Kim J. H., Glant T. T. Anti-CD44 treatment abrogates tissue oedema and leukocyte infiltration in murine arthritis. Nat Med. 1995 Jun;1(6):558–563. doi: 10.1038/nm0695-558. [DOI] [PubMed] [Google Scholar]
  19. Nishida Y., Knudson C. B., Nietfeld J. J., Margulis A., Knudson W. Antisense inhibition of hyaluronan synthase-2 in human articular chondrocytes inhibits proteoglycan retention and matrix assembly. J Biol Chem. 1999 Jul 30;274(31):21893–21899. doi: 10.1074/jbc.274.31.21893. [DOI] [PubMed] [Google Scholar]
  20. Prehm P. Synthesis of hyaluronate in differentiated teratocarcinoma cells. Characterization of the synthase. Biochem J. 1983 Apr 1;211(1):181–189. doi: 10.1042/bj2110181. [DOI] [PMC free article] [PubMed] [Google Scholar]
  21. Recklies A. D., Golds E. E. Induction of synthesis and release of interleukin-8 from human articular chondrocytes and cartilage explants. Arthritis Rheum. 1992 Dec;35(12):1510–1519. doi: 10.1002/art.1780351215. [DOI] [PubMed] [Google Scholar]
  22. Roughley P. J., White R. J. Age-related changes in the structure of the proteoglycan subunits from human articular cartilage. J Biol Chem. 1980 Jan 10;255(1):217–224. [PubMed] [Google Scholar]
  23. Shyjan A. M., Heldin P., Butcher E. C., Yoshino T., Briskin M. J. Functional cloning of the cDNA for a human hyaluronan synthase. J Biol Chem. 1996 Sep 20;271(38):23395–23399. doi: 10.1074/jbc.271.38.23395. [DOI] [PubMed] [Google Scholar]
  24. Spicer A. P., Augustine M. L., McDonald J. A. Molecular cloning and characterization of a putative mouse hyaluronan synthase. J Biol Chem. 1996 Sep 20;271(38):23400–23406. doi: 10.1074/jbc.271.38.23400. [DOI] [PubMed] [Google Scholar]
  25. Spicer A. P., Kaback L. A., Smith T. J., Seldin M. F. Molecular cloning and characterization of the human and mouse UDP-glucose dehydrogenase genes. J Biol Chem. 1998 Sep 25;273(39):25117–25124. doi: 10.1074/jbc.273.39.25117. [DOI] [PubMed] [Google Scholar]
  26. Spicer A. P., McDonald J. A. Characterization and molecular evolution of a vertebrate hyaluronan synthase gene family. J Biol Chem. 1998 Jan 23;273(4):1923–1932. doi: 10.1074/jbc.273.4.1923. [DOI] [PubMed] [Google Scholar]
  27. Spicer A. P., Olson J. S., McDonald J. A. Molecular cloning and characterization of a cDNA encoding the third putative mammalian hyaluronan synthase. J Biol Chem. 1997 Apr 4;272(14):8957–8961. doi: 10.1074/jbc.272.14.8957. [DOI] [PubMed] [Google Scholar]
  28. Tso J. Y., Sun X. H., Kao T. H., Reece K. S., Wu R. Isolation and characterization of rat and human glyceraldehyde-3-phosphate dehydrogenase cDNAs: genomic complexity and molecular evolution of the gene. Nucleic Acids Res. 1985 Apr 11;13(7):2485–2502. doi: 10.1093/nar/13.7.2485. [DOI] [PMC free article] [PubMed] [Google Scholar]
  29. Watanabe K., Yamaguchi Y. Molecular identification of a putative human hyaluronan synthase. J Biol Chem. 1996 Sep 20;271(38):22945–22948. doi: 10.1074/jbc.271.38.22945. [DOI] [PubMed] [Google Scholar]
  30. Weigel P. H., Hascall V. C., Tammi M. Hyaluronan synthases. J Biol Chem. 1997 May 30;272(22):13997–14000. doi: 10.1074/jbc.272.22.13997. [DOI] [PubMed] [Google Scholar]

Articles from Biochemical Journal are provided here courtesy of The Biochemical Society

RESOURCES