Abstract: Objective To investigate the in vitro suppressive effect of antisense oligonucleotides (ASODN) of keratinocyte growth factor receptor (KGFR) on the growth of HaCaT cells induced by keratinocyte growth factor (KGF).Methods HaCaT cells were cultured in vitro with KGF,and treated with various concentrations (0.25,0.5,1.0,2.0,4.0μg/mL) of one of the followings:ASODN1,ASODN2,or sense oligonucleotides (SODN).Cell growth curve assay,MTT assay and colony forming experiments were performed to analyze the effect on proliferation of HaCaT cells.Flow cytometry (FC) was used to investigate the expression of KGFR in the treated HaCaT cells.Results ASODN1 and ASODN2 both suppressed the growth rate of HaCat cells (P<0.05),as shown by the cell growth curve.When >0.25μg/mL ASODN 1 was used,the proliferation induced by KGF was inhibited significantly (P<0.05) as shown by MTT assay,and HaCaT cell clone forming rate was inhibited significantly (P<0.05) as demonstrated by colony forming experiment.Basic KGFR expression in cultured HaCaT cells was inhibited (P<0.05) by ASODN1 in a dose-dependent manner,as shown by flow cytometer analysis.Similar changes were observed for ASODN2.SODN had no suppressive effect on KGFR expression (P>0.05).Conclusion KGFR ASODN is an effective inhibitor of KGFR expression,and could significantly suppress HaCaT cell proliferation mediated by KGF,suggesting that KGFR ASODN might be useful for the treatment of psoriasis.

Key words: Keratinocytes, Receptors,growth factor, Oligonucleotides,antisense, HaCaT cell