外源性信号素3A信号通路对转化生长因子-β1诱导肺癌细胞侵袭、增殖的影响

Effect of semaphorin 3A signaling pathway on invasion and proliferation of lung cancer cells induced by transforming growth factor-β1

目的:探讨外源性信号素3A(Sema 3A)信号通路对转化生长因子-β1(TGF-β1)诱导的肺癌A549细胞侵袭、增殖的影响及可能作用机制。方法:将肺癌A549细胞分为3组,即空白对照组、TGF-β1诱导组(TGF-β1组)、Sema 3A预处理组(Sema 3A+TGF-β1组)。空白对照组细胞正常培养;TGF-β1组加入5μg·L^(-1)TGF-β1;Sema 3A+TGF-β1组首先加入10μmol·L^(-1)Sema 3A,预处理40~50 min后再加入5μg·L^(-1)TGF-β1。检测细胞增殖、侵袭能力和E-cadherin、Akt、P-Akt蛋白表达。结果:Sema 3A+TGF-β1组细胞Sema 3A mRNA相对表达水平显著高于空白对照组和TGF-β1组(均P<0.05)。培养36 h^72 h内,TGF-β1组细胞增殖率显著高于Sema 3A+TGF-β1组和空白对照组(均P<0.05),Sema 3A+TGF-β1组细胞增殖率显著高于空白对照组(均P<0.05)。TGF-β1组穿透滤膜细胞数显著高于Sema 3A+TGF-β1组和空白对照组(均P<0.05),Sema 3A+TGF-β1组穿透滤膜细胞数显著高于空白对照组(P<0.05)。TGF-β1组E-cadherin蛋白表达显著低于Sema 3A+TGF-β1组和空白对照组(均P<0.05),P-Akt蛋白显著高于Sema 3A+TGF-β1组和空白对照组(均P<0.05);Sema 3A+TGF-β1组E-cadherin蛋白表达显著低于空白对照组(P<0.05),P-Akt显著高于空白对照组(P<0.05)。结论:Sema 3A能够抑制TGF-β1诱导肺癌细胞侵袭、增殖的效应,其机制可能与抑制Akt磷酸化、上调E-cadherin表达有关。

Objective： To explore the effect of semaphorin 3A（ Sema 3A） signaling pathway on invasion and proliferation of lung cancer cells induced by transforming growth factor-β1（ TGF-β1） and its mechanisms. Methods：The lung cancer A549 cells were divided into three groups： blank control group（ control）,TGF-β1 induced group（ TGF-β1）,3A Sema pretreatment group（ Sema 3A ＋ TGF-β1）. The control group was cultured with normal cultured,5 μg·L-1TGF-β1 was added in TGF-β1 group,10 μmol·L-1of Sema 3A was firstly added and pretreated for 40-50 min and then 5 μg·L-1of TGF-β1 added in the Sema 3A ＋ TGF-β1 group. Cell proliferation,invasion ability and the expression level of E-cadherin,Akt,P-Akt protein was measured. Results： After Sema 3A pretreatment,the Sema 3A ＋ TGF-β1 group cells were round or oval. Sema 3A mRNA relative expression level in Sema 3A ＋ TGF-β1 group were significantly higher than that in the control group and the TGF-β1 group（ P〈0. 05）. In 36-72 h,the proliferation rate of TGF-β1 group was significantly higher than in Sema 3A ＋ TGF-β1group and the control group（ P〈0. 05）,the proliferation rate in Sema 3A ＋ TGF-β1 group was significantly higher than that in the control group（ P〈0. 05）. The migrated cells number was significantly higher than that in Sema 3A＋ TGF-β1 group and the control group（ P〈0. 05）,migrated cell number in Sema 3A ＋ TGF-β1 group was significantly higher than that in the control group（ P〈0. 05）. The expression of E-cadherin protein was significantly lower than that in Sema 3A ＋ TGF-β1 group and the control group,P-Akt protein was significantly higher than that in Sema 3A ＋ TGF-β1 group and the control group（ P〈0. 05）. E-cadherin protein in Sema 3A ＋ TGF-β1 group was significantly lower than that in the control group,P-Akt protein was significantly higher than that in the control group（ P〈0. 05）. Conclusion： Sema 3A can inhibit invasion and proliferation induced by TGF-β1,the mechanism may be related to the inhibition of Akt phosphorylation and up regulation of E-cadherin expression.