摘要
目的:探究枸橘苷对AGS胃癌细胞抑制作用及其机制。方法:MTT实验检测枸橘苷对AGS细胞活力的影响;流式细胞术分析细胞周期分布以及细胞凋亡;细胞核染色分析AGS细胞在枸橘苷处理之后的形态学变化;Western blot检测枸橘苷对AGS细胞中外源性凋亡通路相关蛋白FasL、caspase-8、caspase-3和PARP,以及内源性凋亡相关蛋白Bak、Bcl-xL、Bax和caspase-9蛋白水平的影响。结果:MTT实验表明枸橘苷抑制AGS胃癌细胞活力,并且呈时间、剂量依赖性(P<0.05);流式细胞术分析结果表明枸橘苷使细胞停滞在G_1期,凋亡数量增加(P<0.05);核染色结果说明,与对照组相比,150μmol/L枸橘苷处理后细胞核明显浓缩,细胞凋亡数量增加;Western blot实验表明枸橘苷上调FasL,激活caspase-8和caspase-3,促使活化的caspase-3底物PARP切割(P<0.05),而对线粒体调节的内源性凋亡通路相关蛋白没有影响。结论:枸橘苷通过激活外源性凋亡通路促使AGs细胞凋亡,抑制胃癌细胞增殖,从而发挥抗肿瘤作用,在临床上可能成为治疗胃癌的新型药物。
AIM： To explore the effect of poncirin on the growth of AGS gastric cancer cells and the underlying mechanism. METHODS： The effect of poncirin on AGS cell viability was measure by MTT assay. The cell cycle distribution and cell apoptosis were analyzed by flow cytometry. Nuclear staining with DAPI was used to reflect the morphological change of the AGS cells treated with poncirin. The protein levels of extrinsic apoptosis pathway-related proteins such as Fas L,caspase-8,caspase-3 and PARP,and mitochondria-mediated intrinsic apoptosis pathway-associated proteins such as Bak,Bcl-x L,Bax and caspase-9 were determined by Western blot. RESULTS： Poncirin inhibited the viability of AGS gastric cancer cells in a time-and concentration-dependent manner（ P 0. 05）. Poncirin induced accumulation of G_1 DNA content and significantly increased total apoptosis in the AGS cells. Nuclear staining showed a dose-dependent increase in the number of apoptotic cells after treated with poncirin. The protein level of Fas L was upregulated in a dose-dependent manner by treatment with poncirin. Poncirin significantly activated caspase-8 and caspase-3. Moreover,poncirin significantly induced the cleavage of PARP in a dose-dependent manner（ P 0. 05）. In addition,the protein levels of Bcl-x L,Bax and Bak were unchanged after treated with different doses of poncirin. Furthermore,caspase-9 was not activated by poncirin treatment in the AGS cells. CONCLUSION： Poncirin has the anti-cancer effect via extrinsic apoptosis pathway to inhibit the growth of AGS gastric cancer cells,possibly making it a therapeutic agent for human gastric cancer treatment.
出处
《中国病理生理杂志》
CAS
CSCD
北大核心
2017年第9期1637-1642,共6页
Chinese Journal of Pathophysiology
关键词
枸橘苷
胃癌细胞
细胞凋亡
Poncirin
Gastric cancer cells
Apoptosis
