启膈散、沙参麦冬汤、通幽汤和补气运脾汤对hEGF刺激的人食管癌EC9706细胞生长信号转导的调节  

Effects of Qigesan,Shashenmaidongtang,Tongyoutang and Buqiyunpitang on hEGF-stimulated proliferation of human esophageal carcinoma EC9706 cells

作　　者：司富春[1] 

机构地区：[1]河南中医学院中医药分子生物学实验室,河南省郑州市450008

出　　处：《世界华人消化杂志》2010年第28期2956-2965,共10页World Chinese Journal of Digestology

基　　金：国家自然科学基金资助项目;No.30371716;No.30873220;No.30973698;国家留学回国人员基金优秀项目;No.国人厅发(2007)170号;国家中医药管理局留学回国人员科技活动择优基金资助项目;No.2003LHR12~~

摘　　要：目的:探讨食管癌中医证方启膈散、沙参麦冬汤、通幽汤和补气运脾汤对hEGF刺激的人食管癌EC9706细胞生长的影响,进而研究该证方抑制食管癌细胞生长信号转导的机制.方法:体外正常条件培养食管鳞癌细胞株EC9706细胞,无血清饥饿细胞24h,用200ng/mLhEGF刺激细胞45min后,加入启膈散、沙参麦冬汤、通幽汤和补气运脾汤,通过镜下形态观察、MTT染色、流式细胞技术观察细胞增殖变化情况.取抑制EGF刺激EC9706细胞增殖IC50所用的各方药物浓度处理细胞,用Westernblot方法检测细胞内PLC-γ1和PI3K介导的生长信号通路的蛋白表达和酪氨酸磷酸水平.结果:启膈散、沙参麦冬汤、通幽汤抑制细胞增殖的IC50值分别为849、1004和1615mg/L,以此浓度干预细胞,可明显抑制细胞生长,阻止细胞进入S期,补气运脾汤对此细胞株无明显抑制作用.在两个生长信号通路中,3方可不同程度地抑制EGFR和PLC-γ1蛋白表达和酪氨酸磷酸化,以及PKCα、MARCKS、PI3K、AKT-1和NF-κBp50蛋白的表达,其中,在PLC-γ1通路中,抑制作用强弱依次是:通幽汤>启膈散>沙参麦冬汤;在PI3K通路中,作用强弱依次是:通幽汤>沙参麦冬汤>启膈散.结论:启膈散、沙参麦冬汤和通幽汤可不同程度地抑制hEGF刺激的EC9706细胞生长.抑制PLC-γ1和PI3K介导的生长信号转导是其抑制EC9706细胞生长的重要机制.AIM：To investigate the effects of Qigesan,Shashenmaidongtang,Tongyoutang and Buqiyunpitang on the proliferation of human esophageal carcinoma EC9706 cells stimulated with human epidermal growth factor （hEGF）,and to explore potential mechanisms involved.METHODS：After cultured EC9706 cells were stimulated with hEGF and then treated with Qigesan,Shashenmaidongtang,Tongyoutang and Buqiyunpitang,respectively,cell morphological changes were observed under an inverted microscope,cell proliferation was determined by MTT assay,cell cycle progression was measured by flow cytometry,and protein expression and tyrosine phosphorylation in PLC-γ1 and PI3Kmediated signaling pathways were determined by Western blot.RESULTS：The half-maximum inhibitory concentrations （IC 50） of Qigesan,Shashenmaidongtang and Tongyoutang were 849,1 004 and 1 615 mg/L,respectively.These drugs at a concentration of IC 50 inhibited hEGF-stimulated cell proliferation and prevented cell cycle progression into S phase.Buqiyunpitang could only weakly inhibit the proliferation of EC9706 cells.Qigesan,Shashenmaidongtang and Tongyoutang inhibited the protein expression and tyrosine phosphorylation of EGFR and PLC-γ1 as well as the protein expression of PKCα,MARCKS,PI3K,AKT-1 and NF-κB p50.Tongyoutang had the strongest inhibitory effects on the PLC-γ1 signaling pathway,followed by Qigesan and Shashenmaidongtang.In contrast,the order of the inhibitory potency for the PI3K signaling pathway was Tongyoutang,Shashenmaidongtang and Qigesan.CONCLUSION：Qigesan,Shashenmaidongtang and Tongyoutang can,to varying degrees,inhibit hEGF-stimulated cell proliferation by suppressing PLC-γ1 and PI3K-mediated signaling pathways.

关 键 词：食管癌 EC9706细胞 人表皮生长因子 启膈散 沙参麦冬汤 通幽汤 补气运脾汤 生长信号转导 

分 类 号：R285.5[医药卫生—中药学]