目的 转化生长因子-β1(TGF-β1)对转移相关基因fascin1表达的作用及机制的研究.方法 以胃癌细胞系MKN45为研究对象,采用蛋白质印迹法检测TGF-β1(10 ng/ml)不同作用时间fascin1表达变化.瞬时转染SMAD2和SMAD4的干扰小RNA(siRNA),检测SMAD依赖信号通路在TGF-β1诱导fascin1表达中的作用.应用ERK、JNK和p38信号通路的特异性阻断药,检测SMAD非依赖途径是否参与TGF-β1诱导fascin1表达.结果 TGF-β1(10 ng/ml)作用MKN45细胞24、48 h后,fascin1表达水平明显高于未处理前(0 h),差异有统计学意义(P﹤0.05).RT-PCR和蛋白质印迹瞬时转染SMAD siRNA,SMAD2和SMAD4基因表达下调,而HK几乎不影响SMAD2和SMAD4基因表达.有效地抑制SMAD2和SMAD4表达后,TGF-β1处理对fascin1表达无明显影响.ERK和JNK信号通路的特异性阻断药PD98095和SP600125处理MKN45细胞后,TGF-β1诱导fascin1表达的作用明显降低;而p38信号通路的特异性阻断药SB203580作用前后,TGF-β1对fascin1表达无明显影响.结论TGF-β1可以通过ERK和JNK信号通路调节转移相关基因fascin1的表达.%Objective To study the effect of transforming growth factor-β1(TGF-β1)on fascin1 expression and clar-ify the mechanism.Method Gastric cancer cell line MKN45 was used in the study,and the effect of TGF-β1(10 ng/ml) with various treatment time on fascin1 expression was detected by Western blot. The effect of SMAD-dependent path-way on TGF-β1-induced fascin1 expression was examined by transient transfection with SMAD2 and SMAD4 siRNA. The effect of SMAD-independent pathway on TGF-β1-induced fascin1 expression was tested using specific inhibitors of ERK,JNK and P38 pathways.Result TGF-β1(10 ng/ml)induced the expression of fascin1 in gastric cancer cell line MKN45 after 24 h and 48 h of treatment as compared with that before treatment(0 h),with statistically significant dif-ference observed (P<0.05). By transient transfection of SMAD siRNA by RT-PCR and western blot, the expression of SMAD2 and SMAD4 were suppressed, while HK was almost not affected by the gene expression of SMAD2 and SMAD4. After effective inhibition of SMAD2 and SMAD4 expression, the TGF-β1-induced fascin1 expression re-mained unchanged. While it was suppressed by the specific inhibitors of JNK and ERK pathways, SP600125 and PD98059 respectively, but not by inhibitors of P38, SB203580. Conclusion TGF-β1 regulates the expression of fas-cin1,by mediating ERK and JNK signaling pathways.
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