HIP1影响食管鳞癌Akt/GSK3β信号通路及EMT相关蛋白表达分析

Effect of HIP1 on Expression of EMT-related Proteins in Esophageal Squamous Cell Carcinoma Through Akt/GSK3β Signaling Pathway

[目的]探讨亨廷顿蛋白相互作用蛋白1(huntingtin interacting protein 1,HIP1)促进食管鳞癌转移的作用及机制。[方法]采用qRT-PCR及Western blot检测Akt抑制剂处理HIP1高或低表达食管鳞癌细胞系中HIP1、GSK3β及EMT标志性分子E-cadherin和Vimentin的表达。采用IHC检测食管鳞癌组织样本中目的分子的表达,并分析其表达相关性。[结果] qRT-PCR和Western blot结果显示:与对照组、shRNA-对照组相比,shRNA-HIP1组细胞中Akt、GSK3β及Vimentin基因和蛋白表达显著降低(P<0.001),E-cadherin基因和蛋白表达显著升高(P<0.001);与对照组、OE-对照组相比,OE-HIP1组细胞中Akt、GSK3β及Vimentin基因和蛋白表达显著升高(P<0.001),E-cadherin基因和蛋白表达显著降低(P<0.001)。IHC及相关性分析结果显示,HIP1与GSK3β、Vimentin表达呈正相关(r=0.336,P<0.001;r=0.561,P<0.001),与E-Cadherin表达呈负相关(r=-0.169,P=0.027);GSK3β与Vimentin表达呈正相关(r=0.317,P<0.001),与E-Cadherin表达呈负相关(r=-0.171,P=0.025)。Akt抑制剂处理HIP1高或低表达食管鳞癌细胞系结果显示,Akt抑制剂处理后E-cadherin表达升高,GSK3β及Vimentin表达降低。[结论]HIP1可能通过激活Akt/GSK3β信号通路促进食管鳞癌EMT的发生。

[Objective] To investigate the effect of huntingtin interacting protein 1(HIP1) on epithelialmesenchymal transition(EMT) related proteins in esophageal squamous cell carcinomas(ESCC) and its mechanism. [Methods] The protein expressions of HIP1,GSK3β,EMT related molecules Ecadherin and Vimentin in cancer tissue samples from 173 ESCC patients were detected by immunohistochemistry(IHC). The mRNA and protein expressions of above indicators were also detected by q RT-PCR and Western blot in ESCC cell lines EC109 and Kyse30,respectively. [Results] IHC and correlation analysis showed that HIP1 expression was positively correlated with GSK3βand Vimentin(r=0.336,P<0.001;r=0.561,P<0.001),and was negatively correlated with E-cadherin expression(r=-0.169,P=0.027).GSK3βexpression was positively correlated with Vimentin expression(r=0.317,P<0.001) and negatively correlated with E-cadherin expression(r=-0.171,P=0.025). The q RT-PCR and Western blot showed that,compared with the control group and the shRNA-control group,the m RNA and protein expressions of Akt,GSK3β and Vimentin in the shRNA-HIP1 group were significantly decreased(P <0.001),while the expression of E-cadherin was significantly increased(P<0.001).Compared with control group and OE-control group,the mRNA and protein expressions of Akt,GSK3β,and Vimentin in OE-HIP1 group were significantly increased(P<0.001),while the expression of E-cadherin were significantly decreased(P<0.001). The expression of Ecadherin was increased,while the expression of GSK3β and Vimentin was decreased in EC109 and Kyse30 cells treated with Akt inhibitor. [Conclusion] HIP1 may promote EMT through activation of Akt/GSK3β signaling pathway in ESCC.