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    糖酵解抑制剂通过抑制糖酵解和促进线粒体调节的途径诱导胃癌细胞凋亡

    Glycolysis inhibitor induces apoptosis of gastric cancer cells by inhibiting glycolysis and promoting mitochondria-regulated pathway

      摘要
    • 摘要:
      目的探究糖酵解抑制剂WZB117通过抑制糖酵解和促进线粒体调节的凋亡途径诱导人胃癌细胞系MGC-803的凋亡的机制。
      方法处于对数生长期的胃癌MGC-803细胞用于实验。根据实验要求，将培养的细胞分为2组：对照组（正常培养的胃癌细胞），WZB117组（用20 μg/mL的葡萄糖运转蛋白抑制剂处理的胃癌细胞）。通过MTS测定试剂盒检测细胞的增殖能力；通过CCK-8测定细胞活力，TUNEL分析细胞细胞凋亡；通过测定ATP含量检测线粒体功能；通过免疫印迹分析Bcl-2、Bax、caspase-3和Cyt-c蛋白和糖酵解相关酶己糖激酶（HK）和磷酸果糖激酶（PFK）蛋白的表达。
      结果24 h时和48 h时WZB117组较对照组细胞增殖降低（P < 0.01）。WZB117组较对照组细胞凋亡率升高（P < 0.01），WZB117组较对照组细胞活力降低（P < 0.01）。12 h时和24 h时WZB117组较对照组ATP含量均降低（P < 0.01）。WZB117组较对照组Bcl-2蛋白表达降低（P < 0.01），WZB117组较对照组Bax、caspase-3和Cyt-c的蛋白表达升高（P < 0.01）。WZB117组较对照组HK和PFK表达降低（P < 0.01）。
      结论WZB117通过抑制糖酵解途径和减少线粒体的ATP产能诱导胃癌细胞系MGC-803的凋亡。

       

      Abstract:
      ObjectiveTo investigate the mechanism of glycolysis inhibitor WZB117-induced apoptosis in human gastric cancer cell line MGC-803 by inhibiting glycolysis and promoting mitochondria-regulated apoptotic pathway.
      MethodsGastric cancer cell line MGC-803 was used in this study, and exponential growth cells were used in the experiment.According to the experimental requirements, the cultured cells were divided into control group(normally cultured cells) and WZB117 group(cells treated with 20 μg/mL glucose transporter inhibitor).The proliferation of cells was detected by MTS kit.Cell viability was measured by CCK-8 and apoptosis was analyzed by TUNEL.Mitochondrial function was detected by ATP content.The expression of Bcl-2, Bax, caspase-3, Cyt-c and glycolysis-related enzymes cells in hexokinase(HK) and cells in phosphofructokinase(PFK) were analyzed by Western blotting.
      ResultsThe proliferation of WZB117 group was lower than that in control group at 24 h and 48 hours(P < 0.01).The apoptosis rate in WZB117 group was higher than that in control group(P < 0.01), and the cell viability in WZB117 group was lower than that in control group(P < 0.01).The ATP content in WZB117 group was lower than that in control group at 12 h and 24 h(P < 0.01).The expression of Bcl-2 protein in WZB117 group was lower than that in control group(P < 0.01), and the expression of Bax, caspase-3 and Cyt-c protein in WZB117 group was higher than that in control group(P < 0.01).The expression of HK and PFK in WZB117 group was lower than that in control group(P < 0.01), indicating that WZB117 could inhibit the expression of enzymes related to glycolysis.
      ConclusionsWZB117 induces apoptosis of gastric cancer cell line MGC-803 by inhibiting glycolysis pathways and reducing mitochondrial ATP production.

       

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