通芪方对胃癌MGC803细胞周期和Wnt/β-catenin信号通路的影响  

Effect of Tongqi Decoction on Cell Cycle and Wnt/β-catenin Signaling Pathway in Gastric Cancer MGC803 Cells

作　　者：贾永森 江春花 韩炳生[2] 曹慧娟 秦丽娟 林清 

机构地区：[1]华北理工大学中医学院,河北唐山063000 [2]深圳市中医院外科,广东深圳518033 [3]华北理工大学基础医学院,河北唐山063000

出　　处：《中华中医药学刊》2016年第5期1126-1129,共4页Chinese Archives of Traditional Chinese Medicine

基　　金：河北省中医药管理局科研计划项目(2015167);深圳市科技研发计划项目(ZYA201007060089A)

摘　　要：目的:从mRNA和蛋白水平阐明通芪方对胃癌MGC803细胞周期和Wnt/β-catenin信号通路的影响作用。方法:MGC803细胞常规体外培养24 h,加入6个质量分数的通芪方,分别于24、48和72 h用MTT染色法检测细胞增殖变化;流式细胞术分析细胞周期;实时荧光定量RT-PCR检测PCNA和Cyclin-D1 mRNA表达变化;western blot法检测Wnt/β-catenin信号通路蛋白表达。结果:通芪方抑制MGC803细胞增殖24 h的IC50浓度约为6.75 g·L^(-1);48 h的IC50浓度约为3.17 g·L^(-1);72 h的IC50浓度约为2.71 g·L^(-1)。通芪方可抑制细胞于G1期,48、72 h时S期细胞比率显著下降,与阴性对照组比较差异有统计学意义(P<0.05);实时荧光定量RT-PCR检测显示48、72 h通芪方组细胞PCNA和Cyclin-D1 mRNA表达水平显著下降,与阴性对照组比较差异有统计学意义(48 h,P<0.05;72 h,P<0.01)。western blot检测显示,通芪方作用细胞72 h,Wnt、β-catenin、PCNA和Cyclin-D1蛋白表达水平显著下降,与阴性对照组比较差异有统计学意义(P<0.05)。结论:通芪方抑制MGC803细胞增殖,具有明显量效、时效性;可调节细胞周期,分子机制与下调Wnt/β-catenin信号通路分子的mRNA和蛋白表达有关。Objective： To illuminate the effect of Tongqi Decoction（ TD） on cell cycle and Wnt/β- catenin signaling pathway in gastric cancer MGC803 cells. Methods： Human MGC803 cells cultured in vitro were treated with TD in 6 concentrations. Cell proliferation was assayed by MTT in 24 h,48 h and 72 h from TD intervention. Cell cycle was determined with flow cytometre at 48 h and 72 h. Concentration of mRNAs of PCNA and Cyclin- D1 was assayed by Real time fluorescence quantitative RT- PCR. Proteins expression in Wnt/β- catenin signaling pathway was measured by Western blot. Results： Half maximal（ 50%） inhibiting concentrations（ IC50） of TD were approximately 6. 75 g·L^（-1）in 24 h,3. 17 g·L^（-1）in 48 h and 2. 71 g·L^（-1）in 72 h. TD blocked MGC803 cells in G1- phase and cell ratios in S- phase decreased drastically compared to that in the negative controlled group（ P〈0. 05）. TD down- regulated mRNA expressions of PCNA and Cyclin- D1 in 48 h,showing statistical significance with those in negative controlled group（ P〈0. 05） and in 72 h,P〈0. 01. Western blot assay showed that TD down- regulated expressions of Wnt,β- catenin,PCNA and Cyclin- D1 in 72 h,showing statistically significance（ P〈0. 05） compared to those in the negative controlled group. Conclusion： TD inhibits MGC803 cells proliferation,taking on dosage- effect and time- effect dependent manner.TD regulating cell cycle has close relation to inhibition of key molecules in Wnt/β- catenin signaling pathway.

关 键 词：通芪方 胃癌 MGC803细胞 细胞周期 WNT/Β-CATENIN 

分 类 号：R285.5[医药卫生—中药学]