角质形成细胞生长因子受体反义寡核苷酸对HaCaT细胞增殖的影响

目的 研究角质形成细胞生长因子受体反义寡核苷酸(KGFR ASODN)对角质形成细胞生长因子(KGF)促HaCaT细胞增殖效应的抑制作用。方法 HaCaT细胞体外培养,采用绘制生长曲线、MTT实验、集落形成实验研究KGFR ASODN对KGF促增殖效应的抑制作用。采用流式细胞仪研究KGFR ASODN对KGFR表达的影响。结果 反义序列1和反义序列2均可抑制HaCaT细胞的生长速度(P<0.05)。高于0.25μg/mL质量浓度反义序列1和反义序列2均可抑制KGF介导的HaCaT细胞增殖(P<0.05)。高于0.25μg/mL质量浓度反义序列1和反义序列2都可抑制KGF诱导的HaCaT细胞集落形成(P<0.05)。HaCaT细胞存在KGFR基础表达,高于0.25μg/mL质量浓度反义序列1和反义序列2均可抑制HaCaT细胞KGFR表达(P<0.05),抑制率呈浓度依赖性,正义序列核苷酸(S-ODN)对KGFR表达无抑制作用(P>0.05)。结论 KGFR ASODN可有效抑制KGFR表达,进而抑制KGF介导的HaCaT细胞过度增殖。

The effects of antisense oligonucleotides of keratinocyte growth factor receptor on proliferation of HaCaT cells

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.