MT-3通过激活NF-κB信号通路抑制食管癌病理进展

MT-3 aggravates pathological process of esophageal cancer through activation of NF-κB signaling passway

摘要:

目的  食管癌的发生与多种基因突变相关，基因的表达异常产生肿瘤，本文集中探讨食管癌的发生发展机制。方法  选取河北医科大学附属邢台人民医院2009年7月-2011年2月肿瘤科食管癌患者92例。选取其肿瘤组织及其癌旁组织样本，检测金属硫蛋白3（MT-3）在食管癌组织和其癌旁组织中的表达，在食管癌细胞系中构建过表达MT-3和低表达MT-3的细胞系，检测这些MT-3表达不同时，NF-κB信号通路在其中是否处于激活状态。采用Western blot检测P65、pP65蛋白的表达，免疫荧光检测pP65是否在MT-3过表达时有入核表达。结果  定量聚合酶链反应（qPCR）结果显示食管癌肿瘤组织中MT-3的表达高于癌旁组织（P =0.001）。通过qPCR检测构建的高表达和低表达MT-3的食管癌细胞系均成功构建，进而检测P65、pP65，显示在过表达MT-3时，pP65蛋白表达增多，低表达MT-3时pP65蛋白表达减少，P65则显现出相反的结果，且表达均有统计学意义，所以认为MT-3高表达时NF-κB信号激活。免疫荧光检测高表达MT-3的食管癌细胞时，细胞核入核明显增多，进一步说明MT-3激活了NF-κB信号通路。 结论  MT-3在食管癌中通过激活NF-κB信号通路进而抑制食管癌的病理进程。

Abstract:

Objective To research the development mechanism of esophageal cancer. Methods Ninety-two cases of esophageal cancer patients treated in Xingtai Pepole's Hospital of Hebei Medical University from July 2009 to February 2011 were selected for the study. MT-3 expression was detected in esophageal cancer tissues and the peri-cancerous tissues. MT-3 overexpression and low-expression cell lines were constructed. The activation state of NF-κB signaling pathway was checked at different levels of MT-3 expression. Then the expressions of P65 and pP65 were tested by Western blot. Immunofluorescence was used to further test whether pP65 was expressed in the nuclei when MT-3 was over-expressed. Results The expression of MT-3 in the tumor tissues was higher than that in the adjacent tissues (P = 0.001). A high MT-3 expression esophageal cancer cell line and a low MT-3 expression cell line were successfully constructed. pP65 protein expression was enhanced when MT-3 was over-expressed, but decreased at low MT-3 expression; in contrast, P65 showed the opposite results with significant differences, which suggested NF-κB signal was activated when MT-3 was over expressed. Immunofluorescence revealed that in MT-3 overexpression cells, more pP65 was expressed in the nuclei, which further supported that MT-3 activated NF-κB signaling pathway. Conclusions MT-3 suppresses the pathological process of esophageal cancer through activation of NF-κB signals.