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Differential effect of growth factors on hyaluronan synthase gene expression in fibroblasts exposed to oxidative stress

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Abstract

The aim of this study was to evaluate how growth factors (PDGF-BB, EGF, and TGF-1β) modulate hyaluronan synthase (HAS) activities in normal or stressed cultured human skin fibroblasts. The effects of concomitant treatment with cytokines and FeSO4 plus ascorbate on HAS mRNA expression, protein synthesis, and hyaluronic acid (HA) concentrations were also studied. Treatment of fibroblasts with growth factors up-regulated HAS gene expression and increased HAS enzymes and HA production. PDGF-BB induced HAS mRNA expression, protein synthesis, and HA production more efficiently than EGF and TGF-1β. EGF was less effective than TGF-1β. In addition, TGF-1β reduced the expression and synthesis of HAS3, while PDGF-BB and EGF had the opposite effect. Concomitant treatment with growth factors and the oxidant was able to further increase HAS mRNA expression, once again with the exception of HAS3 with TGF-1β. HAS protein synthesis was reduced, while HA levels were unaffected in comparison to those obtained from exposure to FeSO4 plus ascorbate alone. In conclusion, although growth factors plus the oxidant synergistically induced HAS mRNA expression in part, enzyme production was not correlated with this increase. Moreover, the increase in HAS mRNA levels was not translated into a consequent rise in HA concentration.

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Abbreviations

DMEM:

Dulbecco’s modified Eagle’s medium

ECM:

extracellular matrix

EGF:

epidermal growth factor

FBS:

fetal bovine serum

GAGs:

glycosaminoglycans

HA:

hyaluronic acid

HASs:

hyaluronan synthases

HRP:

horseradish peroxidase

IFN-γ:

interferon-γ

IL-1β:

interleukin-1β

NADH:

reduced nicotinamide adenine dinucleotide

PBS:

phosphate-buffered saline

PCR:

polymerase chain reaction

PDGF-BB:

platelet derived growth factor BB

PGs:

proteoglycans

ROS:

reactive oxygen species

SDS:

sodium dodecyl sulfate

TBS:

tris-buffered solution

TGF-1β:

transforming growth factor 1β

TNF-α:

tumor necrosis factor α

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Authors and Affiliations

  1. Department of Biochemical, Physiological, and Nutritional Sciences, School of Medicine, University of Messina, Policlinico Universitario, Torre Biologica, 5° piano, Via C. Valeria, 98125, Messina, Italy

    G. M. Campo, A. Avenoso, S. Campo, A. D’Ascola & A. Calatroni

  2. Department of Morphology, Biochemistry, Physiology and Animal Production, School of Veterinary Medicine, University of Messina, Contrada Annunziata, 98168, Messina, Italy

    A. M. Ferlazzo

Authors
  1. G. M. Campo
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  2. A. Avenoso
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  3. S. Campo
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  4. A. D’Ascola
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  5. A. M. Ferlazzo
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  6. A. Calatroni
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Correspondence to G. M. Campo.

Additional information

Published in Russian in Biokhimiya, 2007, Vol. 72, No. 9, pp. 1196–1206.

Originally published in Biochemistry (Moscow) On-Line Papers in Press, as Manuscript BM07-118, July 29, 2007.

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Campo, G.M., Avenoso, A., Campo, S. et al. Differential effect of growth factors on hyaluronan synthase gene expression in fibroblasts exposed to oxidative stress. Biochemistry Moscow 72, 974–982 (2007). https://doi.org/10.1134/S0006297907090088

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  • DOI: https://doi.org/10.1134/S0006297907090088

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