目的 研究白藜芦醇在体外对胶质瘤细胞株U251的作用,并探讨其潜在的调控机制.方法 选取处于对数生长期U251细胞分为空白对照组和不同剂量(25 mol/L、50 mol/L、100 mol/L)白藜芦醇组.采用四甲基噻唑蓝(MTT)比色法评估白藜芦醇对U251细胞增殖率的影响;流式细胞术检测白藜芦醇对U251细胞凋亡的影响;逆转录聚合酶链反应法(RT-PCR) 检测白藜芦醇对U251细胞Wnt、β-catenin、p53、p21、Bcl-2和Bax信使RNA (mRNA)表达水平影响;免疫蛋白印迹法(Western blot)检测白藜芦醇对U251细胞Wnt、β-catenin、p53、p21、Bcl-2和Bax蛋白表达水平影响.结果 白藜芦醇作用U251细胞72 h后,MTT检测结果显示空白对照组U251细胞增殖率为(94±5)%,而不同剂量(25 mol/L、50 mol/L、100 mol/L)白藜芦醇组U251细胞增殖率分别为(64±4)%、(43±4)%和(27±2)%,较空白对照组明显降低,差异均具有统计学意义(P<0.05);流式细胞术检测结果显示空白对照组U251细胞凋亡率为(4±1)%,而不同剂量(25 mol/L、50 mol/L、100 mol/L)白藜芦醇组U251细胞凋亡率分别为(25±2)%、(55±3)%、(72±5)%,较空白对照组明显增高,差异均具有统计学意义(P<0.05);RT-PCR和Western blot检测结果显示与空白对照组比较,不同剂量白藜芦醇组U251细胞Wnt、β-catenin、p53、p21和Bcl-2表达水平均明显降低,差异均具有统计学意义(P<0.05);而不同剂量白藜芦醇组与空白对照组比较,U251细胞Bax表达水平明显增高,差异均具有统计学意义(P<0.05).结论 白藜芦醇可诱导人胶质瘤细胞株U251细胞增殖抑制和凋亡,其作用机制可能与抑制Wnt/β-catenin/p53信号通路活化相关.%Objective To study the effect of resveratrol on glioma cell line U251 in vitro and its potential regu-latory mechanism.Methods U251 cells in logarithmic growth phase were divided into the blank control group and dif-ferent doses of(25 mol/L,50 mol/L,100 mol/L)resveratrol groups.MTT colorimetric method was used to evaluate the effects of resveratrol on proliferation rate of U251 cells;the effect of resveratrol on apoptosis of U251 cells were detect-ed by flow cytometry;reverse transcriptase polymerase chain reaction(RT-PCR)and Western blot were used to detect the effect of resveratrol on the mRNA and protein expression levels of U251 cells on Wnt,β-catenin,p53,p21,Bcl-2 and Bax,respectively.Results After resveratrol treating U251 cells for 72 h,the results of MTT showed that U251 cell proliferation rate of the blank control group was(94±5)%,and the U251 cell proliferation rates of the different doses of resveratrol groups were(64±4)%,(43±4)% and(27±2)%,respectively.Compared with the blank control group,the cell proliferation rates of the resveratrol groups were significantly decreased,and there were statistical significance between the different doses of resveratrol groups and the blank control group(P<0.05);the results of flow cytometry showed that the apoptosis rate of U251 cells in the blank control group was(4±1)%,and the apoptosis rate of U251 cells in the differ-ent doses of resveratrol groups were(25±2)%,(55±3)% and(72±5)%,respectively.The apoptosis rates of the different doses of resveratrol groups were significantly higher than that in the blank control group,and there were statistically sig-nificant differences between the different doses of resveratrol group and the blank control group(P<0.05);RT-PCR and Western blott showed that,compared with the blank control group,the different doses of resveratrol groups had lower ex-pression of Wnt,β-catenin,p53,p21 and Bcl-2 in U251 cells and higher expression of Bax(P<0.05).Conclusion Res-veratrol can induce the proliferation inhibition and apoptosis of human glioma cells lines U251,which is associated with suppressing Wnt/β-catenin/p53 signal pathway.
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