【摘要】 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。更多还原

【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 ker- atinocyte 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 in- vestigate the expression of KGFR in the treated HaCaT cells.Results ASODN1 and ASODN2 both sup- pressed 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 A- SODN is an effective inhibitor of KGFR expression,and could significantly suppress HaCaT cell prolifera- tion mediated by KGF,suggesting that KGFR ASODN might be useful for the treatment of psoriasis.更多还原