中文摘要：

目的：通过研究噎膈血瘀证、脾气虚证患者及健康人血清对食管癌 EC9706细胞增殖、磷脂酰肌醇3-激酶（PI3K）/蛋白激酶B（Akt）/核因子kappaB（NF-κB）信号通路相关分子mRNA及蛋白表达的调控，探讨噎膈血瘀证、脾气虚证的分子机制。方法本研究于2014年6—12月选择来源于唐山市人民医院、迁安燕山医院肿瘤内科的病理确诊的食管鳞癌患者。按照《中华人民共和国中医药行业标准———中医病症诊断疗效标准》确定噎膈血瘀证或脾气虚证标准，选取血瘀证、脾气虚证患者各10人，另选取健康志愿者10人（来自两院体检人员）。将EC9706细胞于37℃，5％饱和湿度的二氧化碳（ CO2）培养箱孵育24 h，饥饿24 h，分别加入倍比梯度的噎膈血瘀证、脾气虚证患者和健康人血清，噻唑蓝（MTT）染色法检测细胞增殖变化；实时荧光定量聚合酶链反应（PCR）检测PI3K、Akt、NF-kB表达；免疫印迹（Western blot）法检测PI3K/Akt/NF-κB信号通路相关分子蛋白表达。结果血瘀证患者血清刺激细胞的50％增殖率为71.1μl/ml，脾气虚证为118.0μl/ml；血瘀证患者血清上调细胞 PI3K、Akt 和NF-κB mRNA表达水平，与各对照组比较差异有统计学意义（ P﹤0.05），脾气虚证患者血清无上调mRNAs作用；血瘀证患者血清促进细胞表皮生长因子受体（EGFR）、PI3K、Akt、磷酸化的蛋白激酶B（p-Akt）和 NF-κB等蛋白表达水平增高，与各对照组比较差异有统计学意义（ P﹤0.05），脾气虚证患者血清无促进此五蛋白表达作用。结论噎膈血瘀证患者血清能够促进细胞增殖，该证候的分子机制可能与PI3K/Akt/NF-κB信号通路过度激活有关；脾气虚证患者血清无刺激细胞增殖作用，其证候分子机制有待探讨。

英文摘要：

Objective To investigate the molecular mechanisms of Blood-Stagnation Syndrome（ BSS）and spleen-Qi-dificiency syndrome（ SQDS）patients with Esophageal cancer（ EC）through observation on cells proliferation,mRNAs and proteins expression in PI3K/Akt/NF-κB signaling pathway regulated by sera from patients. Methods From June to December, 2014,we enrolled patients who were definitely diagnosed with esophageal squamous cell carcinoma in oncology department of Tangshan People＇s Hospital and Yanshan Hospital. According to TCM Professional Standard of People＇s Republic of China/ TCM diagnosis and efficacy standard,we determined the standard of blood stasis syndrome and spleen-yang deficiency syndrome in patients with dysphagia. We enrolled 10 patients of blood stasis syndrome,10 patients with spleen-yang deficiency syndrome, and another 10 healthy controls（people who received physical examination from the two hospitals）. EC9706 cells were put into CO2 incubator with a temperature of 37℃ and a saturation humidity of 5% and were cultured for 24 hours and kept hungury for 24 hours. Then serum of patients of blood stasis syndrome and spleen-yang deficiency syndrome and healthy controls was put into the＆nbsp;incubator by proportional gradient,and MTT staining method was used to detect the changes in cell proliferation;the expression levels of PI3K,Akt and NF-κB mRNA were measured by Real-time PCR,and the protein expression of relevant molecules of PI3K/Akt/NF-κB signalling pathway was detected by Western blot method. Results Half maximal （ 50%） promotion concentration（PC50）of BSS was 71. 1 μl/ml,PC50 of SQDS was 118. 0 μl/ml. Sera from BSS patients promoted PI3K,Akt and NF-κB mRNAs expression,being significantly different from those in control group（P ﹤0. 05）,however,sera from SQDS patients didn＇t work. Western blot assay showed that BSS patients＇sera stimulated over-expression of EGFR,PI3K,Akt, p-Akt and NF-κB,showing significant difference from control group（P﹤0. 05）,howeve