六君子汤提取物抑制食管癌Ec9706细胞增殖及其对信号转导和转录活化因子3信号通路的影响  

Inhibition of Extract of Liujunzi Decoction on Ec9706 Cells Growth of Esophageal Carcinoma and Its Effects on Signaling Pathway of Signal Transducers and Activations of Transcripition 3

作　　者：崔姗姗[1] 高小玲[1] 王慧慧[1] 吴耀松[1] 尹素改[1] 陈玉龙[1] 

机构地区：[1]河南中医药大学,河南省郑州市450046

出　　处：《中国全科医学》2016年第24期2948-2952,共5页Chinese General Practice

基　　金：国家自然科学基金资助项目(81173177);河南省教育厅自然科学基金资助项目(15B360003)

摘　　要：目的研究六君子汤提取物对食管癌Ec9706细胞增殖的抑制作用及其对信号转导和转录活化因子3(STAT3)信号通路的影响。方法于2013—2015年,采用70%乙醇提取六君子汤,正丁醇、乙酸乙酯及水提法过夜萃取,用MTT法观察提取物对Ec9706细胞活性的影响,选取最优提取部位,采用酶联免疫吸附试验(ELISA)法观察六君子汤乙酸乙酯部位对Ec9706细胞分泌相关细胞因子〔包括白介素6(IL-6)、转化生长因子β1(TGF-β1)、血管内皮生长因子(VEGF)〕的影响,采用Western-blotting法测定六君子汤乙酸乙酯部位对食管癌Ec9706细胞STAT3和磷酸化STAT3(p-STAT3)蛋白表达的影响。结果对照组、六君子汤70%乙醇部位、六君子汤正丁醇部位、六君子汤水提部位Ec9706细胞OD值比较,差异有统计学意义(F=448.90,P<0.01)。药物处理组对Ec9706细胞生长的抑制效果依次为:六君子汤正丁醇部位>六君子汤70%乙醇部位>六君子汤水提部位。六君子汤乙酸乙酯部位Ec9706细胞OD值比较,差异有统计学意义(F=19.78,P<0.001)。在250μg/ml时,六君子汤70%乙醇部位、六君子汤正丁醇部位、六君子汤水提部位对Ec9706细胞的抑制率分别为7.31%、23.24%、-25.07%,而在200μg/ml时,六君子汤乙酸乙酯部位对Ec9706细胞的抑制率为77.03%,抑制效果明显优于其他部位,故筛选的有效部位为六君子汤乙酸乙酯部位。应用概率法计算六君子汤乙酸乙酯部位抑制率IC35、IC50、IC70,代表低、中、高浓度,相应的浓度为63.08、93.00、133.70μg/ml。对照组、六君子汤低浓度组、六君子汤中浓度组IL-6、TGF-β1、VEGF水平比较,差异均有统计学意义(P<0.01);其中六君子汤低浓度组、六君子汤中浓度组IL-6、TGF-β1、VEGF水平低于对照组(P<0.05)。对照组、六君子汤低浓度组、六君子汤中浓度组STAT3、p-STAT3表达水平比较,差异均有统计学意义(P<0.05);其中六君子汤低浓度组、六君子汤中浓度组STAT3Objective To observe the inhibition of extract of Liujunzi decoction on esophageal carcinoma cells Ec9706 growth and the effects on signaling pathway of signal transducers and activations of transcripition 3（STAT3）. Methods From 2013 to 2015,Liujunzi decoction was extracted by 70% ethanol,overnight extraction was made by normal butanol,ethyl acetate and water extraction. MTT method was applied to observe the influence of the extracts on the activity of Ec9706 cells. The optimal extracting parts,and enzyme linked immunosorbent assay（ ELISA） was used to observe the effects of ethyl acetate extract in Liujunzi decoction on the related cytokines〔 including interleukin - 6（ IL-6）,transforming growth factor - β1（ TGF-β1）, vascular endothelial growth factor（VEGF）〕secreted by Ec9706 cells. Western - blotting method was adopted to test the effects of ethyl acetate extract in Liujunzi decoction on protein expression of esophageal carcinoma cells Ec9706 STAT3 and phosphorylation STAT3（p-STAT3）. Results There was significant difference in OD value of Ec9706 cells among the control group,70%ethanol parts,normal butanol parts,and water - extraction parts of Liujunzi decoction（ F = 448. 90,P ﹤ 0. 01 ） . The inhibition effects of Ec9706 cell growth in all medication groups were shown as follows：normal butanol parts of Liujunzi decoction﹥ 70% ethanol parts of Liujunzi decoction ﹥ water - extraction part of Liujunzi decoction. There was significant difference in OD value of Ec9706 cells in ethyl acetate parts of Liujunzi decoction（ F = 19. 78,P ﹤ 0. 001）. When the concentration was 250μg/ ml,the inhibition ratio of 70% ethanol parts,normal butanol parts,and water - extraction parts of Liujunzi decoction on Ec9706 were 7. 31% ,23. 24% ,and - 25. 07% respectively,while the concentration was 200 μg/ ml,the inhibition rate of the ethyl acetate extract of Liujunzi decoction on Ec9706 cells was 77. 03% ,showing a inhibitory effect that was significantly better than the other parts,so the

关 键 词：食管肿瘤 六君子汤 提取物 乙酸乙酯 STAT3 

分 类 号：R735.1[医药卫生—肿瘤]