title: Effect of mitomycin on normal dermal fibroblast and HaCat cell: an in vitro study.
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 addition, mitomycin can inhibit HaCat cell proliferation, so epithelial cell needs more protecting to avoid mitomycin's side effect when it is applied clinically.
authors.affiliation: Otolaryngology-Head and Neck Surgery Department, Ningbo First Hospital, Ningbo 315000, China.
authors.forename: Yao-wen
authors.initials: YW
authors.lastname: Wang
authors.forename: Ji-hao
authors.initials: JH
authors.lastname: Ren
authors.forename: Kun
authors.initials: K
authors.lastname: Xia
authors.forename: Shu-hui
authors.initials: SH
authors.lastname: Wang
authors.forename: Tuan-fang
authors.initials: TF
authors.lastname: Yin
authors.forename: Ding-hua
authors.initials: DH
authors.lastname: Xie
authors.forename: Li-hua
authors.initials: LH
authors.lastname: Li
mesh_terms: D002460:Cell Line; D049109:Cell Proliferation / Q000187:drug effects; D003094:Collagen / Q000378:metabolism*; D004305:Dose-Response Relationship, Drug; D005347:Fibroblasts / Q000187:drug effects / Q000378:metabolism*; D036341:Intercellular Signaling Peptides and Proteins / Q000378:metabolism*; D015603:Keratinocytes / Q000187:drug effects / Q000378:metabolism*; D016685:Mitomycin / Q000008:administration & dosage*; D019384:Nucleic Acid Synthesis Inhibitors / Q000494:pharmacology; D012333:RNA, Messenger / Q000378:metabolism*; D015398:Signal Transduction / Q000187:drug effects / Q000502:physiology
chemical_list: D036341:Intercellular Signaling Peptides and Proteins; D019384:Nucleic Acid Synthesis Inhibitors; D012333:RNA, Messenger; D016685:Mitomycin; D003094:Collagen