根皮素诱导胃癌HGC-27细胞凋亡的机制研究

Effects of phloretin on cell proliferation and apoptosis in gastric cancer cell line HGC-27 cells

[目的]观察根皮素对胃癌HGC-27细胞增殖、凋亡的影响,并探讨其可能的作用机制。[方法]体外用不同浓度根皮素处理HGC-27细胞,CCK-8比色法检测药物处理后细胞活力;PI/Annexin V双染法检测细胞凋亡率;免疫印迹法检测细胞凋亡相关蛋白Bax、Bcl-2及COX-2、EP1蛋白表达;ELISA法检测细胞上清液中前列腺素E2(PGE_(2))含量。[结果]CCK-8比色法结果显示,根皮素处理后可抑制HGC-27细胞的增殖,呈剂量、时间依赖性;根皮素处理后,PI/Annexin V双染后流式细胞仪检测结果显示,随着药物浓度的增加,HGC-27细胞凋亡率也逐渐增加;免疫印迹结果显示,随着药物浓度的增加,HGC-27细胞凋亡相关蛋白Bax蛋白表达增加,且Bcl-2蛋白的表达降低,同时COX-2、EP1蛋白表达降低;ELISA结果显示,随着药物浓度的增加,HGC-27细胞上清液中PGE_(2)含量逐渐降低。[结论]根皮素在体外能抑制胃癌HGC-27细胞增殖并促进其凋亡,可能的机制是通过下调COX-2表达、PGE_(2)合成及EP1表达,激活线粒体凋亡途径。

[Objective]To observe the effect of phloretin on cell proliferation and apoptosis in gastric cancer cell line HGC-27 cells and explore the possible mechanisms.[Methods]HGC-27 cells were treated with phloretin,and the proliferaton of HGC-27 cells was measured by CCK-8 assay.Apoptosis was detected by PI/Annexin V stainning.Western blot were used for COX-2,EP1,Bcl-2 and Bax protein analysis.The expression of PGE_(2)in the culture medium was detected by ELISA.[Results]From the data of CCK-8,the cell proliferation of HGC-27 cells was inhibited by phloretin in a dose-dependent and time-dependent manner.After treated with phloretin,the apoptosis rate of HGC-27 cells increased.The treatment with phloretin promoted the expression of pro-apoptotic factor Bax,and suppressed the expression of anti-apoptotic factor Bcl-2.And the expressions of COX-2 and PGE_(2)are suppressed.[Conclusion]Phloretin inhibits the cell proliferation and induces cell apoptosis in HGC-27 cells,which is associated with a reduction in the levels of COX-2,PGE_(2)and EP1,thus mediating the mitochondrial pathway.