曲古抑菌素A通过调控蛋白激酶C促进食管鳞癌细胞迁移

Trichostatin A promotes the migration of esophageal squamous carcinoma cells by protein kinase C signaling pathway

目的探讨组蛋白去乙酰化酶抑制剂曲古抑菌素A(TSA)对人食管鳞癌细胞(ESCC)迁移能力的影响及其可能的作用机制。方法培养食管鳞癌细胞KYSE-150和EC9706;Transwell法检测TSA、蛋白激酶C(PKC)抑制剂AEB071对食管鳞癌细胞迁移的影响;观察TSA、PKC抑制剂AEB071对人食管鳞癌细胞形态学的影响;Western blotting检测E-钙黏蛋白(E-cadherin)、波形蛋白(vimentin)、β-连环蛋白(β-catenin)、Slug、ac H3和H3蛋白的表达水平。结果TSA促进食管鳞癌细胞的迁移,PKC抑制剂AEB071部分抑制TSA促进食管鳞癌细胞的迁移作用;TSA促使细胞由类椭圆形的上皮样细胞形态向类似梭形的成纤维细胞样形态的上皮-间质转化(EMT),AEB071抑制TSA诱导的细胞形态学变化;TSA处理后E-cadherin蛋白表达水平降低,vimentin、β-catenin、Slug、ac H3表达水平升高,联合应用TSA与AEB071,TSA诱导的上述EMT相关蛋白水平的变化得到部分抑制,使E-cadherin蛋白表达水平增加,vimentin、β-catenin、Slug表达水平降低。结论TSA通过促进食管鳞癌细胞EMT增加其迁移能力,其机制可能由PKC相关信号通路介导。

Objective To investigate the effect of histone deacetylase inhibitor trichostatin A(TSA)on the migration of human esophageal squamous carcinoma cells(ESCC)and the possible mechanism.Methods KYSE-150 cells and EC9706 cells were cultured and Transwell assay was performed to detect the role of TSA alone and combined with protein kinase C(PKC)inhibitor AEB071 on cell migration;the images of morphology after cells treatment with TSA or combination of AEB071 with TSA;Western blotting was conducted to examine the protein level of epithelial-mesenchymal transition(EMT)related signaling molecules.Results TSA promoted the migration of ESCC cells significantly,and PKC inhibitor AEB071 partly inhibited the effect of TSA-promoted ESCC cells migration.Treatment with TSA resulted in the cell morphology transitioned from epithelia oval-like to mesenchymal spindle-like,indicating the EMT.AEB071 partially rescued ESCC cells morphological changes which TSA induced.Western blotting showed that TSA reduced the expression of E-cadherin and augmented the expression of vimentin,β-catenin,Slug and ac H3,whereas AEB071 obviously blocked the EMT-related protein level changes which induced by TSA.Conclusion TSA promotes ESCC cells migration via inducing EMT process and the mechanism may be mediated by PKC signaling pathway.