根皮素对人胃癌细胞系SGC-7901的影响及作用机制

Effect of phloretin on human gastric cancer cell line SGC-7901 and its mechanism

目的探讨根皮素对人胃癌细胞系SGC-7901的影响及其作用机制。方法培养人胃癌细胞系SGC-7901,分别采用0.3%DMSO(DMSO组)、顺铂2.5 mg/L(DDP组)、辣椒素受体1(VR1)拮抗剂辣椒平(Cap)0.01 mmol/L(Cap组)以及根皮素40、80、160μg/mL处理细胞,采用Cap 0.01 mmol/L预处理1 h后加根皮素160μg/mL(Cap+根皮素组)处理细胞,同时设置阴性对照组不做任何处理。MTT法检测细胞活性,流式细胞术检测细胞凋亡,通过JC-1荧光染色评估线粒体膜电位(MMP),Western blot法检测凋亡相关蛋白表达。结果与DMSO组和阴性对照组相比,根皮素40、80、160μg/mL处理12、24、48 h后,细胞存活率降低,且呈剂量-时间依赖性(P<0.05);与DDP组比较,根皮素80、160μg/mL处理12、24、48 h后,细胞存活率降低(P<0.05);Cap+根皮素组细胞存活率较根皮素160μg/mL组增加(P<0.05)。与DMSO组和阴性对照组比较,根皮素40、80、160μg/mL处理后,细胞凋亡率增加,且呈剂量依赖性(P<0.05),根皮素160μg/mL组MMP降低,Bcl-2蛋白表达降低,Bax、Caspase-3前体、Caspase-3和细胞色素C蛋白表达增加(P<0.05)。与根皮素160μg/mL组相比,Cap+根皮素组细胞凋亡率降低,MMP升高,Bcl-2蛋白表达增加,Bax、Caspase-3前体、Caspase-3和细胞色素C蛋白表达降低(P<0.05)。结论根皮素可以降低SGC-7901细胞存活率,诱导细胞凋亡;其作用机制可能与线粒体凋亡途径和VR1有关。

Objective To explore the effect of phloretin on human gastric cancer cell line SGC-7901 and its mechanism.Methods Human gastric cancer cell line SGC-7901 was treated with 0.3%DMSO(group DMSO),cis-platinum 2.5 mg/L(group DDP),vanilloid receptor 1(VR1)antagonist Capsazepine(Cap)0.01 mmol/L(group Cap),and phloretin 40,80,160μg/mL.The cells were pretreated with Cap 0.01 mmol/L for 1 hour and then treated with phloretin 160μg/mL(group Cap+phloretin).The cells without any treatment was taken as the group C.The cell viability was determined by MTT assay,cell apoptosis was detected by flow cytometry,mitochondrial membrane potential(MMP)was evaluated by JC-1 fluorescence staining,and apoptosis-related proteins were detected by Western blot.Results Compared to groups of DMSO and C,the cell survival rate was decreased in a dose-time dependent manner after treated with phloretin 40,80,160μg/mL for 12,24 and 48 hours(P<0.05).Compared with group DDP,the cell survival rate was decreased after treated with phloretin 80 and 160μg/mL for 12,24 and 48 hours(P<0.05).The cell survival rate was increased in group Cap+phloretin than that in group phloretin 160μg/mL(P<0.05).Compared with groups of DMSO and C,the cell apoptosis rate was increased in a dose-dependent manner after treated with phloretin 40,80,160μg/mL,and MMP was decreased,Bcl-2 protein expression was decreased,protein expressions of Bax,Caspase-3 precursor,Caspase-3 and cytochrome C were increased in group phloretin 160μg/mL(P<0.05).Compared with group phloretin 160μg/mL,the apoptosis rate was decreased,MMP was increased,and the Bcl-2 protein expression was increased,and protein expressions of Bax,Caspase-3 precursor,Caspase-3 and cytochrome C were decreased in group Cap+phloretin(P<0.05).Conclusion Phloretin can decrease the survival rate of SGC-7901 cells and induce cell apoptosis.Its mechanism may be related to mitochondrial apoptosis pathway and VR1.