J Korean Med Sci.  2011 Mar;26(3):417-424. 10.3346/jkms.2011.26.3.417.

Transcriptional Regulation of Proteoglycans and Glycosaminoglycan Chain-synthesizing Glycosyltransferases by UV Irradiation in Cultured Human Dermal Fibroblasts

• Shin JE
• Oh JH
• Kim YK
• Jung JY
• Chung JH

• KMID: 2157881
• DOI: http://doi.org/10.3346/jkms.2011.26.3.417

Abstract

Various kinds of glycosaminoglycans (GAGs) and proteoglycans (PGs) have been known to be involved in structural and space-filling functions, as well as many physiological regulations in skin. To investigate ultraviolet (UV) radiation-mediated regulation of GAGs and PGs in cultured human dermal fibroblasts, transcriptional changes of many types of PGs and GAG chain-synthesizing enzymes at 18 hr after 75 mJ/cm2 of UV irradiation were examined using quantitative real-time polymerase chain reaction methods. Hyaluronic acid synthase (HAS)-1, -2, and -3 and hyaluronidase-2 mRNA expressions were significantly increased by UV irradiation. Expressions of lumican, fibromodulin, osteoglycin, syndecan-2, perlecan, agrin, versican, decorin, and biglycan were significantly decreased by UV irradiation, while syndecan-1 was increased. Expressions of GAG chain-synthesizing glycosyltransferases, xylosyltransferase-1, beta1,3-glucuronyltransferase-1, beta1,4-galactosyltransferase-2, -4, exostosin-1, chondroitin polymerizing factor, and chondroitin sulfate synthase-3 were significantly reduced, whereas those of beta1,3-galactosyltransferase-6, beta1,4-galactosyltransferase-3, -7, beta-1,3-N-acetylglucosaminyltran sferase-2, and -7 were increased by UV irradiation. Heparanase-1 mRNA expression was increased, but that of heparanase-2 was reduced by UV irradiation. Time-course investigation of representative genes showed consistent results. In conclusion, UV irradiation may increase hyaluronic acid production through HAS induction, and decrease other GAG productions through downregulation of PG core proteins and GAG chain-synthesizing glycosyltransferases in cultured human dermal fibroblasts.

Keyword

Glycosaminoglycans; Proteoglycans; Glycosyltransferases; Ultraviolet Radiation; Dermal Fibroblasts

MeSH Terms

Cell Line
Fibroblasts/metabolism/radiation effects
Gene Expression Regulation/radiation effects
Glucuronosyltransferase/genetics/radiation effects
Glycosaminoglycans/*biosynthesis/chemistry
Glycosyltransferases/genetics/*metabolism
Humans
Hyaluronic Acid/biosynthesis
Hyaluronoglucosaminidase/genetics/radiation effects
Polymerase Chain Reaction
Proteoglycans/*biosynthesis/genetics/radiation effects
RNA, Messenger/analysis/genetics
Skin/*metabolism/radiation effects
Transcription, Genetic/radiation effects
*Ultraviolet Rays

Figure

• 

  Fig. 1 Fold changes of UV-induced mRNA expression of HA-related genes and various proteoglycans (PGs) in cultured human dermal fibroblasts. Human dermal fibroblasts were incubated for 18 hr after 75 mJ/cm2 of UV irradiation. Total RNA was isolated from UV-irradiated or sham-irradiated cells, converted to the cDNA, and applied to the quantitative real-time polymerase chain reaction experiments for each target genes. (A) Changes of HA-related genes. (B) Changes of KSPGs. (C) Changes of HSPGs. (D) Changes of CS/DSPGs. Values are mean fold changes ± standard deviation (SD) (n = 3 or 4). *P < 0.05 versus control sham-irradiated cells. N.D. means not detected in both control and UV-irradiated cells. HAS, hyaluronic acid synthase.

• 

  Fig. 2 Fold changes of UV-induced mRNA expression of GAG chain-synthesizing glycosyltransferases in cultured human dermal fibroblasts. Human dermal fibroblasts were incubated for 18 hr after 75 mJ/cm2 of UV irradiation. Total RNA was isolated from UV-irradiated or sham-irradiated cells, converted to the cDNA, and applied to the quantitative real-time polymerase chain reaction experiments for each target genes. (A) Changes of GAG chain-synthesizing common glycosyltransferases. (B) Changes of KS chain-synthesizing glycosyltransferases. (C) Changes of HS chain-synthesizing glycosyltransferases and heparanases. (D) Changes of CS and DS chain-synthesizing glycosyltransferases. Values are mean fold changes ± SD (n = 3 or 4). *P < 0.05 versus control sham-irradiated cells. N.D. means not detected in both control and UV-irradiated cells. B3GALT, β1,3-galactosyltransferase; B4GALT, β1,4-galactosyltransferase; B3GAT, β1,3-glucuronyltransferase; B3GNT, β-1,3-N-acetylglucosaminyltransferase; CHPF, chondroitin polymerizing factor; CHSY, chondroitin sulfate synthase; CSGALNACT, chondroitin sulfate N-acetylgalactosaminyltransferase.

• 

  Fig. 3 Time-dependent fold changes of UV-induced mRNA expression of HA-related genes, proteoglycans, and GAG chain-synthesizing glycosyltransferases in cultured human dermal fibroblasts. Human dermal fibroblasts were incubated for 6, 12, 18, or 24 hr after 75 mJ/cm2 of UV irradiation. Total RNA was isolated from UV-irradiated or sham-irradiated cells, converted to the cDNA, and applied to the quantitative real-time polymerase chain reaction experiments for each target genes. (A) Changes of HAS2, osteoglycin, syndecan-1, perlecan, and decorin. (B) Changes of xylosyltransferase-1, B3GNT1, B4GALT2, exostosin-1, and CHPF. Values are mean fold changes ± SD (n = 3 or 4). *P < 0.05 versus control sham-irradiated cells at each time point.

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