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目的:探讨花椒毒素对人胃癌SGC-7901细胞体外增殖抑制作用,并从死亡受体途径对其进行机制研究。方法:不同浓度花椒毒素作用于胃癌SGC-7901细胞48h后,MTT法检测花椒毒素对胃癌SGC-7901细胞的增殖抑制作用;Hoechst 33258染色及Annexin V-FITC/PI双染法观察花椒毒素诱导细胞凋亡的形态学变化和早中晚期凋亡的区别;流式细胞术检测死亡受体及其配体Fas/Fas L、DR5/TRAIL及Bid蛋白,Caspase-8检测试剂盒检测Caspase-8活性。结果:花椒毒素呈剂量依赖性抑制SGC-7901细胞的增殖,IC50为75.6μg/ml;花椒毒素作用后贴壁细胞减少,凋亡小体的数量增加,且呈剂量依赖性;给药48h后,DR5/TRAIL的表达升高,在一定浓度范围内Fas/Fas L蛋白的表达升高,Caspase-8的活性增强,Bid蛋白活化增多。结论:花椒毒素通过上调DR5/TRAIL的表达,在一定浓度范围内上调Fas/Fas L蛋白的表达,启动死亡受体途径,促进Caspase-8的活化,进而激活下游效应因子;活化的Caspase-8可激活Bid蛋白,进入线粒体发挥作用,可能是花椒毒素诱导SGC-7901细胞凋亡的途径之一。
Objective: To investigate the inhibitory effect of xanthotoxin on the proliferation of human gastric cancer cell line SGC-7901 in vitro and to study its mechanism from the death receptor pathway. Methods: The proliferation of SGC-7901 cells was determined by MTT assay after different concentrations of pricklyashin (SGC-7901) cells were treated with different concentrations of pricklyashin for 24 hours. Hoechst 33258 staining and Annexin V-FITC / PI double staining were used to observe the effects of prionotoxin- Apoptotic morphological changes and early, middle and late apoptosis; Fas / Fas L, DR5 / TRAIL and Bid proteins were detected by flow cytometry; Caspase-8 activity was detected by Caspase-8 assay kit . Results: Xanthoxins inhibited the proliferation of SGC-7901 cells in a dose-dependent manner with an IC50 of 75.6μg / ml. After treated with xanthotoxin, the number of adherent cells decreased and the number of apoptotic bodies increased in a dose-dependent manner. After 48h , The expression of DR5 / TRAIL was increased. The expression of Fas / Fas L protein increased, the activity of Caspase-8 increased and the activity of Bid protein increased in a certain concentration range. CONCLUSIONS: Xanthotoxin can up-regulate the expression of Fas / Fas L protein and activate the death effector pathway by up-regulating the expression of DR5 / TRAIL in a certain concentration range, and then activate the downstream effector factor. The activation of Caspase-8 Activates Bid protein into mitochondria and may be one of the pathways by which toxin SGC-7901 can induce apoptosis of SGC-7901 cells.