[Effect of nitric oxide on HaCaT cell migration].
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.
Yang
Shi-wei
SW
Institute of Burn Research, the Third Military Medical University, Chongqing 400038, China.
Wu
Jun
J
Luo
Gao-xing
GX
Zhang
Xiao-rong
XR
Hu
Xiao-hong
XH
Peng
Yan-meng
YM
Yang
Jun-jie
JJ
Luo
Xiao-li
XL
Wang
Ying
Y
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
D012333:RNA, Messenger; D009569:Nitric Oxide; D020742:rhoA GTP-Binding Protein