摘要
目的探讨趋化因子受体CXCR4(CXC chemokine receptor 4,CXCR4)在食管癌上皮间质转化(epithelialmesenchymal transition,EMT)发生中的作用及其参与食管癌侵袭转移的可能机制。方法化学合成靶向CXCR4的siRNA,脂质体作用下转染食管癌EC9706细胞,同时以转染阴性对照siRNA、单纯转染脂质体和空白细胞作为对照。RT-PCR和Western blot检测CXCR4 siRNA对CXCR4 mRNA和蛋白表达的抑制作用;侵袭小室、划痕实验和MTT检测CXCR4 siRNA对EC9706细胞侵袭转移和增殖能力的影响;RT-PCR和Western blot检测CXCR4 siRNA对E-cadherin和vimentin mRNA和蛋白表达的影响。结果靶向CXCR4 siRNA可以有效抑制食管癌EC9706细胞中CXCR4 mRNA和蛋白的表达,与对照组相比,差异有统计学意义(P<0.01),且有时间依赖性(P<0.05);转染CXCR4 siRNA的EC9706细胞侵袭转移和增殖能力较对照组显著下降(P<0.05);转染CXCR4 siRNA的EC9706细胞随着CXCR4表达下降,E-cadherin表达随之升高,而vimentin表达则相应下降,差异有统计学意义(P<0.05)。结论 CXCR4可通过促进食管癌EMT的发生,参与其侵袭转移过程。
Objective To study the role of CXCR4 in epithelial-mesenchymal transition （EMT） in esophageal carcinoma and the mechanism of CXCR4 participating in the invasion and metastasis of esophageal carcinoma. Methods siRNA targeting CXCR4 was chemically synthesized and was transfected into esophageal carcinoma EC9706 cells. EC9706 cells transfected with negative siRNA or Lipofectamine 2000 and vacant cells were used as controls. CXCR4 mRNA and protein levels were evaluated by semi-quantitative RT-PCR and Western blotting after 24, 48 and 72 h. Invasion and metastasis ability and proliferation ability were detected by Boyden chamber assay, wound-healing assay and MTT assay. The mRNA and protein levels of E-cadherin and vimentin in EC9706 cells transfected with CXCR4 siRNA were tested by semi-quantitative RT-PCR and Western blotting. Results siRNA targeting CXCR4 could suppress the expression of CXCR4 in EC9706 cells efficiently, as compared to that in control groups （P〈0.01）, in a time-dependent manner （P〈0.05）. The invasion and metastasis ability and proliferation ability of EC9706 cells transfected with CXCR4 siRNA were decreased greatly as compared to those in control groups （P〈0.05）. With the decrease of CXCR4 expression, the expression of E-cadherin was increased while the expression of vimentin was decreased （P〈0.05）. Conclusion CXCR4 participates in the invasion and metastasis of esophageal carcinoma through EMT.
出处
《第三军医大学学报》
CAS
CSCD
北大核心
2013年第22期2443-2447,共5页
Journal of Third Military Medical University
基金
河南省基础与前沿技术研究计划(132300410127)
河南省教育厅科学技术研究重点项目(13A320638)~~
