吲哚美辛诱导的胃癌细胞凋亡与Wnt/β-连接蛋白信号通路及其下游凋亡调控蛋白的关系

摘要

Objective To investigate the relationship between Wnt/p-Catenin signal pathway including apoptosis regulatory proteins (Bcl-2, Caspase-3) downstream and the apoptosis of gastric cancer cell induced by indomethacin. Method The SGC7901 cells were cultured with indomethacin for 24 hours at the final concentration of 50, 100 and 200 u.mol/L, the control group without indomethacin intervention was used. The proliferation activity of cells was tested by CCK8 method. The apoptosis rate of SGC7901 cells in smears was measured via TUNEL method. Intracellular expressions of P-Catenin, Bcl-2 and Caspase-3 proteins were assessed by Western blot technique. Results The survival rate of SGC7901 cell was (92.71 + 4.81)% after intervention of indomethacin for 24 hours at the concentration of 50 jtmol/L. The survival rates of cells under the effect of indomethacin of 100 and 200 fimol/L were decreased significantly. Apoptosis rate in control group was 0, the same parameter was increased significantly after administration of indomethacin. The highest expression of P-Catenin and Bcl-2 proteins and the minimum amount of Caspase-3 protein were found in the control group. After intervention of indomethacin, the expressions of P-Catenin and Bcl-2 reduced while the amount of Caspase-3 rose significantly, which were concentration-dependent obviously. Conclusions Wnt/p-Catenin signal pathway plays an important role in the apoptosis of SGC7901 cell induced by indomethacin. Its downstream proteins Bcl-2 and Caspase-3 were also involved in the regulation of this process.%目的:探讨Wnt/β-连接蛋白(Catenin)信号通路及其下游凋亡调控蛋白Bcl-2和Caspase-3的表达与吲哚美辛引起的胃癌细胞凋亡的关系.方法:用终浓度为50、100、200 μmol/L的吲哚美辛干预SGC7901细胞24 h,并设不加吲哚美辛的对照组.用CCK8法检测细胞增殖活性,用TUNEL法检测涂片中胃癌细胞的凋亡率,用Western blot法检测细胞内β-Catenin、Bcl-2和Caspase-3蛋白的表达.结果:SGC7901细胞在50 μmol/L的吲哚美辛作用24 h后细胞存活率为(92.71±4.81)％,100和200 μmol/L浓度组的细胞存活率则明显降低.对照组细胞的凋亡率为0,吲哚美辛干预后的细胞的凋亡率明显升高.对照组β-Catenin和Bcl-2蛋白表达最高而Caspase-3表达最低.吲哚美辛干预后β-Catenin和Bcl-2的表达明显降低,Caspase-3表达明显增高,且表现出明显的浓度依赖性.结论:Wnt/β-Catenin信号通路在吲哚美辛引导的SGC7901细胞凋亡过程中发挥重要的作用,其下游调控蛋白Bcl-2、Caspase-3共同参与了这一过程的调控.