Journal of Zhejiang University SCIENCE B

, Volume 13, Issue 12, pp 997–1005 | Cite as

Effect of mitomycin on normal dermal fibroblast and HaCat cell: an in vitro study

  • Yao-wen Wang
  • Ji-hao Ren
  • Kun Xia
  • Shu-hui Wang
  • Tuan-fang Yin
  • Ding-hua Xie
  • Li-hua Li
Article

Abstract

Objective

To evaluate the effects of mitomycin on the growth of human dermal fibroblast and immortalized human keratinocyte line (HaCat cell), particularly the effect of mitomycin on intracellular messenger RNA (mRNA) synthesis of collagen and growth factors of fibroblast.

Methods

The normal dermal fibroblast and HaCat cell were cultured in vitro. Cell cultures were exposed to 0.4 and 0.04 mg/ml of mitomycin solution, and serum-free culture medium was used as control. The cellular morphology change, growth characteristics, cell proliferation, and apoptosis were observed at different intervals. For the fibroblasts, the mRNA expression changes of transforming growth factor (TGF)-β1, basic fibroblast growth factor (bFGF), procollagen I, and III were detected by reverse transcription polymerase chain reaction (RT-PCR).

Results

The cultured normal human skin fibroblast and HaCat cell grew exponentially. A 5-min exposure to mitomycin at either 0.4 or 0.04 mg/ml caused marked dose-dependent cell proliferation inhibition on both fibroblasts and HaCat cells. Cell morphology changed, cell density decreased, and the growth curves were without an exponential phase. The fibroblast proliferated on the 5th day after the 5-min exposure of mitomycin at 0.04 mg/ml. Meanwhile, 5-min application of mitomycin at either 0.04 or 0.4 mg/ml induced fibroblast apoptosis but not necrosis. The apoptosis rate of the fibroblast increased with a higher concentration of mytomycin (p<0.05). A 5-min exposure to mitomycin at 0.4 mg/ml resulted in a marked decrease in the mRNA production of TGF-β1, procollagen I and III, and a marked increase in the mRNA production of bFGF.

Conclusions

Mitomycin can inhibit fibroblast proliferation, induce fibroblast apoptosis, and regulate intracellular protein expression on mRNA levels. In additon, mitomycin can inhibit HaCat cell proliferation, so epithelial cell needs more protecting to avoid mitomycin’s side effect when it is applied clinically.

Key words

Mitomycin Fibroblast HaCat cell Apoptosis TGF-β1 bFGF Procollagen 

CLC number

R762 

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Copyright information

© Zhejiang University and Springer-Verlag Berlin Heidelberg 2012

Authors and Affiliations

  • Yao-wen Wang
    • 1
  • Ji-hao Ren
    • 2
  • Kun Xia
    • 3
  • Shu-hui Wang
    • 2
  • Tuan-fang Yin
    • 2
  • Ding-hua Xie
    • 2
  • Li-hua Li
    • 2
  1. 1.Otolaryngology-Head and Neck Surgery DepartmentNingbo First HospitalNingboChina
  2. 2.Otolaryngology-Head and Neck Surgery Department, the Second Xiangya HospitalCentral South UniversityChangshaChina
  3. 3.State Key Laboratory of Medical GeneticsCentral South UniversityChangshaChina

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