title: [Effect of nitric oxide on HaCaT cell migration].
abstract: OBJECTIVE
  To investigate the effect of exogenous nitric oxide (NO) on the migration of HaCaT cell and its possible mechanism.
  
  
  METHODS
  Sodium nitroprusside (SNP) was used as the donor of NO. Different concentrations of SNP (0.1, 1.0, 10.0, 100.0, 1000.0 micromol/L) were added into nutrient culture medium of HaCaT cells. Cell migration rate was observed and calculated at post scratching hour (PSH) 0 (immediately after scratching), 6, 12, 24, 48. The most suitable concentration of SNP and culture duration were selected as stimulation condition. Cytoskeletons of HaCaT cells were observed under confocal laser scanning microscope. The expressions of integrin beta 1, RhoA, Rac1 and Cdc42 of cells in experiment group (cultured with 10.0 micromol/L SNP for 24 hours) and negative control group were determined at mRNA and protein levels with RT-PCR and Western blot respectively. Data were processed with one-way analysis of variance (ANOVA) and repeated measure ANOVA.
  
  
  RESULTS
  Migration rate of HaCaT cells in each group increased gradually as time after scratching went on. There were significant differences between PSH 6-48 and PSH 0 in cells cultured with 10.0 micromol/L SNP (F = 31.002, P values all below 0.05). Pili were rarely observed in negative control group with slender stress fibers in cells. In comparison, the amount of pili amount increased obviously in experiment group with thickened stress fibers. Compared with those of cells in control group (RhoA protein expression = 0.64 +/- 0.04), integrin beta 1 expression decreased obviously (F = 8.25, P = 0.015), RhoA (0.92 +/- 0.04), Cdc42 and Rac1 were up-regulated at both protein (with F value respectively 7.25, 14.10, 6.50, P values all below 0.05) and mRNA levels (with F value respectively 23.67, 10.39, 9.52, P values all below 0.05).
  
  
  CONCLUSIONS
  Exogenous NO in suitable concentration can promote the proliferation and migration of HaCaT cell, suggesting it exerts significant effect in wound repair. The changed cytoskeletons and the down-regulated integrin beta 1 expression may be involved in this process.
authors: Institute of Burn Research, the Third Military Medical University, Chongqing 400038, China.
authors: Shi-wei
authors: SW
authors: Yang
authors: Jun
authors: J
authors: Wu
authors: Gao-xing
authors: GX
authors: Luo
authors: Xiao-rong
authors: XR
authors: Zhang
authors: Xiao-hong
authors: XH
authors: Hu
authors: Yan-meng
authors: YM
authors: Peng
authors: Jun-jie
authors: JJ
authors: Yang
authors: Xiao-li
authors: XL
authors: Luo
authors: Ying
authors: Y
authors: Wang
mesh_terms: D002460:Cell Line; D002465:Cell Movement / Q000187:drug effects*; D003599:Cytoskeleton / Q000187:drug effects* / Q000378:metabolism; D006801:Humans; D009569:Nitric Oxide / Q000494:pharmacology*; D012333:RNA, Messenger / Q000235:genetics; D020742:rhoA GTP-Binding Protein / Q000235:genetics / Q000378:metabolism
chemical_list: D012333:RNA, Messenger; D009569:Nitric Oxide; D020742:rhoA GTP-Binding Protein