二氢青蒿素通过抑制SIRT1的表达而增强5-氟尿嘧啶对胃癌的抗肿瘤活性

发布时间：2018-03-16 00:13

本文选题：二氢青蒿素　切入点：SIRT蛋白　出处：《中国病理生理杂志》2017年12期 　论文类型：期刊论文

【摘要】：目的:探讨二氢青蒿素对5-氟尿嘧啶治疗胃癌的辅助作用并研究其机制。方法:实验分为对照组、二氢青蒿素组、5-氟尿嘧啶组、5-氟尿嘧啶联合二氢青蒿素组和5-氟尿嘧啶+二氢青蒿素+SIRT1质粒组。MTT法检测胃癌细胞系BGC-823在5-氟尿嘧啶联合二氢青蒿素处理下的细胞活力。Western blot实验检测5-氟尿嘧啶联合二氢青蒿素对BGC-823细胞SIRT1和NADPH氧化酶表达水平,caspase-9和caspase-3活化水平及凋亡信号调节激酶1(ASK1)和c-Jun氨基末端激酶(JNK)蛋白磷酸化水平的影响。流式细胞术检测BGC-823细胞在5-氟尿嘧啶和二氢青蒿素联合处理下的活性氧簇(ROS)生成水平和细胞凋亡率。结果:二氢青蒿素处理能显著抑制BGC-823细胞SIRT1的表达并增加NADPH氧化酶的蛋白水平,明显提高BGC-823细胞对5-氟尿嘧啶的敏感性,降低5-氟尿嘧啶的半数抑制浓度;转染SIRT1表达质粒后,二氢青蒿素联合5-氟尿嘧啶对BGC-823细胞的杀伤活性受到显著抑制(P0.05)。二氢青蒿素能明显促进5-氟尿嘧啶对BGC-823细胞生成ROS的诱导效应和ASK1及JNK的磷酸化(P0.05)。用ROS清除剂N-乙酰半胱氨酸(NAC)或JNK特异性抑制剂SP600125处理后,二氢青蒿素联合5-氟尿嘧啶对BGC-823细胞的杀伤活性和caspase-9及caspase-3的活化均受到明显抑制(P0.05)。另外,NAC能显著抑制二氢青蒿素联合5-氟尿嘧啶对JNK磷酸化的促进作用,而SP600125却不能影响BGC-823细胞ROS的产生,表明JNK是ROS的下游分子。结论:二氢青蒿素联合5-氟尿嘧啶通过SIRT1/NADPH氧化酶/ROS/JNK通路诱导胃癌细胞发生caspase依赖的凋亡。
[Abstract]:Objective: To investigate the effect of Dihydroartemisinin on 5- assisted two fluorouracil in the treatment of gastric cancer and to study its mechanism. Methods: the experiment was divided into control group, two dihydroartemisinin group, 5- fluorouracil group, 5- fluorouracil combined with dihydroartemisinin group and two 5- fluorouracil + two dihydroartemisinin +SIRT1 plasmid group.MTT was detected in gastric cancer cell line BGC-823 cells.Western blot in the two experiment 5- fluorouracil combined with dihydroartemisinin treatment to detect 5- fluorouracil two Dihydroartemisinin on BGC-823 cells SIRT1 and NADPH oxidase expression levels of caspase-9 and caspase-3 activation and apoptosis signal regulating kinase 1 (ASK1) and c-Jun N-terminal kinase (JNK) protein phosphorylation. ROS detection of BGC-823 cells flow cytometry in two 5- fluorouracil and dihydroartemisinin combined treatment under (ROS) generation and apoptosis. Results: two dihydroartemisinin treatment can significantly To inhibit the expression of BGC-823 SIRT1 cells and increased the protein level of NADPH oxidase, significantly improve the sensitivity of BGC-823 cells to 5- fluorouracil, lower half inhibitory concentration of 5- fluorouracil; transfection of SIRT1 expression plasmid, the killing activity of two dihydroartemisinin combined with 5- fluorouracil on BGC-823 cells was significantly inhibited (P0.05) two dihydroartemisinin can significantly. To promote 5- fluorouracil on formation of BGC-823 cells induced by ROS effect and ASK1 and the phosphorylation of JNK (P0.05) by ROS. N- scavenger N-acetylcysteine (NAC) or JNK inhibitor SP600125 treatment, two activation of dihydroartemisinin combined with 5- fluorouracil on the cytotoxicity of BGC-823 cells and caspase-9 and caspase-3 were significantly inhibited (P0.05). In addition, NAC can significantly inhibit the two dihydroartemisinin combined with 5- fluorouracil on the phosphorylation of JNK role, while SP600125 did not affect BGC-823 cell R The generation of OS indicates that JNK is a downstream molecule of ROS. Conclusion: two, artemisinin combined with 5- fluorouracil induces SIRT1/NADPH dependent apoptosis in gastric cancer cells through SIRT1/NADPH oxidase /ROS/JNK pathway.

【作者单位】：湖州市中心医院药学部;湖州市中心医院放疗介入科;
【分类号】：R735.2

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