MTH1抑制剂TH287对胃癌细胞BGC-823的抑制作用及其机制

Inhibitory effect of MTH1 inhibitor TH287 on gastric cancer BGC-823 cells and its mechanism

目的观察MTH1小分子抑制剂对胃癌细胞株BGC-823的作用及其机制。方法体外培养胃癌细胞株BGC-823,用不同浓度MTH1抑制剂TH287干预BGC-823细胞。CCK-8法和克隆形成实验检测BGC-823细胞增殖;划痕实验和Transwell实验检测BGC-823细胞迁移能力;流式细胞仪和线粒体膜电位检测细胞凋亡水平;Western blot检测凋亡相关蛋白Bcl-2、Bax及PI3K/AKT信号通路相关蛋白表达水平;AKT磷酸化激活剂SC79预处理细胞验证TH287对PI3K/AKT信号通路的作用。结果CCK-8检测结果显示,TH287抑制BGC-823细胞活力,效果呈剂量和时间依赖性(P<0.05)。克隆形成实验结果显示,随着药物浓度的增加,细胞克隆形成数减少。划痕实验和Transwell实验结果显示,随着TH287浓度增加,BGC-823细胞迁移能力下降(P<0.05)。Annexin-Ⅴ/PI双染实验和线粒体膜电位检测结果显示,TH287诱导BGC-823细胞凋亡增加和线粒体膜电位下降(P<0.05);Western blot结果显示,TH287诱导凋亡相关蛋白Bcl-2/Bax比值下降(P<0.05),并且促进PI3K表达增加,AKT磷酸化水平上升(P<0.05);AKT磷酸化激活剂SC79预处理胃癌细胞发现,TH287对胃癌的抑制作用减弱,细胞凋亡减少,细胞活力增加。结论MTH1抑制剂TH287能够抑制BGC-823细胞增殖及迁移,并且可能通过调控PI3K/AKT信号通路调控BGC-823细胞凋亡。

Objective To investigate the effect of MTH1 small molecule inhibitor on gastric cancer cell line BGC-823 and its mechanism.Methods Gastric cancer cell line BGC-823 cells were cultured in vitro and treated with different concentrations of MTH1 inhibitor TH287.Cell proliferation was detected by CCK-8 and clone formation assay.The effect of TH287 on the migration ability of BGC-823 cells was detected by scratch assay and Transwell assay.Apoptosis was determined by flow cytometry and mitochondrial membrane potential.Bcl-2,Bax and PI3K,p-AKT expression levels were detected by Western blot.AKT phosphorylation activator SC79 pretreated BGC-823 cells to verify the effect of TH287 on the PI3K/AKT signaling pathway.Results The results of CCK-8 assay showed that TH287 inhibited cell viability of BGC-823 cells in a concentration-and time-dependent manner(P<0.05).Clony formation results showed that the number of clone cells decreased with the increase concentration of TH287.Wound healing and Transwell assay results showed that TH287 inhibited the ability of migration in BGC-823 cells(P<0.05).The results of Annexin-Ⅴ/PI double staining and mitochondrial membrane potential assay showed that TH287 induced BGC-823 cells apoptosis and mitochondrial membrane potential collapse(P<0.05).The results of Westetn blot revealed that the ratio of Bcl-2/Bax and the expression levels of PI3K and p-AKT decreased after the treatment of TH287(P<0.05).After pretreatment with AKT phosphorylation activator SC79,TH287 decreased the inhibitory effect on gastric cancer,decreased apoptosis and increased cell vitality.Conclusion MTH1 inhibitor TH287 can inhibit BGC-823 cell proliferation and migration.Moreover,TH287 can induce BGC-823 cell apoptosis via regulating PI3K/AKT pathway.