摘要
目的探讨白藜芦醇(Res)对胃腺癌细胞(AGS)生长的影响及其与Wnt/β-catenin信号通路的关系。方法体外培养胃癌细胞株AGS,并以不同浓度Res(25、50、100μmol/L)干预24、48、72 h后,四甲基偶氮唑蓝(MTT)法测定细胞抑制率;不同浓度Res干预24 h流式细胞仪检测细胞周期,细胞免疫组化检测Wnt1、β-catenin、Lgr5的表达情况。实验数据以x±s表示,采用SPSS13.0软件进行统计分析,多组间比较采用单因素方差分析,两两比较采用t检验。P<0.05表示差异有统计学意义。结果不同浓度Res(25、50、100μmol/L)干预24、48、72 h后,AGS细胞的状态变差,细胞形态变圆密度及数量减少,贴壁较差,细胞增殖抑制率分别为:25μmol/L Res作用24 h组(0.2033±0.0207)、48 h组(0.3949±0.0199)、72 h组(0.5102±0.0155);50μmol/L Res作用24 h组(0.3554±0.0207)、48 h组(0.5157±0.0321)、72 h组(0.6167±0.0248);100μmol/L Res作用24 h组(0.5005±0.0199)、48 h组(0.6251±0.0299)、72 h组(0.7271±0.0147),细胞增殖抑制率呈时间依赖性与剂量依赖性(P<0.05);不同浓度Res作用24 h后,检测G1期细胞所占比分别为空白组(48.33±0.21)%、25μmol/L Res作用组(52.61±0.41)%、50μmol/L Res作用组(58.53±0.48)%、100μmol/L Res作用组(71.16±0.20)%,细胞周期阻滞在G0/G1期(P<0.05);不同浓度Res干预24 h后100μmol/L组(207.36±0.52)、50μmol/L组(202.65±0.53)、25μmol/L组(197.13±1.07)Wnt1灰度值均高于对照组(191.40±0.28)(P<0.05);100μmol/L组(206.51±0.68)、50μmol/L组(200.49±0.30)、25μmol/L组(196.53±0.59)β-catenin灰度值均高于对照组(191.98±0.30)(P<0.05),100μmol/L组(209.22±0.51)、50μmol/L组(204.53±0.92)、25μmol/L组(195.7±60.90)Lgr5灰度值均高于对照组(188.16±0.86)(P<0.05)。结论 Res可抑制胃癌细胞AGS的生长作用,其机制可能通过减少Wnt/β-catenin信号通路的活化而产生的。
Objective To investigate the influence of resveratrol(Res)on the growth of gastric cancer cells(AGS)in vitro and the underlying mechanism.Methods AGS cells were cultured with Res at different concentrations(25,50,and100μmol/L)for24,48,or72h.MTT assay was used to determine the reduced growth of AGS cells at24,48,or72h after treatment.Flow cytometry was used to detect the cell cycle arrest at24h.Immunocytochemistry was used to detect the expression of Wnt1,beta-catenin,and Lgr5at24h.Results After AGS cells were treated with different concentrations of Res(25,50,and100μmol/L)for different durations,the cells became round in morphology,decreased in density,and showed poorer adherence.Res inhibited cell proliferation in a time and dose dependent manner(25μmol/L:0.2033±0.0207at24h,0.3949±0.0199at48h,0.5102±0.0155at72h;50μmol/L:0.3554±0.0207at24h,0.5157±0.0321at48h,0.6167±0.0248at72h;100μmol/L:0.5005±0.0199at24h,0.6251±0.0299at48h,0.7271±0.0147at72h;P<0.05).Res arrested cells in G1phase in a concentration dependent manner(percentage of G1phase cells in control cells:48.33±0.21;25μmol/L:52.61±0.41;50μmol/L:58.53±0.48;100μmol/L:71.16±0.20,P<0.05).Res altered the expression of Wnt1(100μmol/L:207.36±0.52;50μmol/L:202.65±0.53;25μmol/L:197.13±1.07;control:191.40±0.28;P<0.05),beta-catenin(100μmol/L:206.51±0.68;50μmol/L:200.49±0.30;25μmol/L:196.53±0.59;control:191.98±0.30;P<0.05),and Lgr5(100μmol/L:209.22±0.51;50μmol/L:204.53±0.92;25μmol/L:195.7±60.90;control:188.16±0.86;P<0.05)in a concentration dependent manner.Conclusion Res inhibits the growth of AGS cells probably by regulating the Wnt/beta-catenin signaling pathway.
作者
张振华
侯海金
张斌
闫慧明
Zhang Zhenhua;Hou Haijin;Zhang Bin;Yan Huiming(Shanxi Medical University, Taiyuan030001, China;Department of General Surgery, the Second Hospital of Shanxi Medical University, Taiyuan030001, China)
出处
《中华临床医师杂志(电子版)》
CAS
2017年第15期2087-2092,共6页
Chinese Journal of Clinicians(Electronic Edition)