DHA联合5-FU对人胃癌细胞增殖的抑制及线粒体呼吸链膜蛋白复合体的表达变化  

The effect of combination of docosahexaenoic acid and 5-fluorouracil on the proliferation of human gastric cancer cell line AGS and the expression of mitochondrial respiratory membrane protein complexes

作　　者：王曙逢[1] 赵志浩[1] 刘竹君[1] 高琨[2] 李幼芬[2] 

机构地区：[1]西安交通大学医学部第一附属医院普通外科,陕西西安710061 [2]西安交通大学生命科学与技术学院,陕西西安710049

出　　处：《西安交通大学学报（医学版）》2015年第4期491-495,共5页Journal of Xi’an Jiaotong University（Medical Sciences）

摘　　要：目的：探讨二十二碳六烯酸（DHA）联合5-氟尿嘧啶（5-FU）对体外培养人胃腺癌细胞株 AGS 增殖的抑制作用并初步分析其机制。方法以不同梯度浓度的 DHA、5-FU 作用于体外培养的 AGS 细胞,采用中效原理和 Chou-Ta-lalay 联合指数原理计算药物单用和合用时的中效浓度（IC50）和合用指数（CI）,确定药物合适作用浓度。实验分为对照组、DHA 组、5-FU 组和 DHA 联合5-FU 组,四甲基偶氮唑蓝（MTT）观察细胞增殖抑制率,流式细胞仪检测细胞周期变化,Western blot 分析细胞线粒体呼吸链膜蛋白复合体的表达变化。结果DHA、5-FU 单用可时间-剂量依赖地抑制 AGS 细胞增殖（P 〈0.05）。DHA、5-FU 单用24、48 h 时 IC50分别为51.60、34.82μg/mL（DHA）和45.90、16.86μg/mL（5-FU）。DHA 与5-FU 合用时,DHA 可显著增强5-FU 对 AGS 细胞生长增殖的抑制效应,并将5-FU的 IC50减少3.56~2.15倍,CI〈1二者呈现协同作用。与对照组、DHA 组、5-FU 组比较,联合干预组 AGS 细胞的DNA 合成前期（G0/Gl 期）细胞分布比例明显增加,DNA 合成期（S 期）细胞分布比例明显减少（P 〈0.05）。与对照组、DHA 组、5-FU 组比较,联合干预组 AGS 线粒体呼吸链膜蛋白复合体表达显著下降（P 〈0.05）。结论DHA 和5-FU 合用可协同抑制人胃腺癌细胞株 AGS 的生长增殖,可能通过抑制线粒体呼吸链膜蛋白复合体表达而干扰细胞能量代谢及阻滞细胞周期于 G0/G1期。Objective To investigate the inhibitory effects of docosahexaenoic acid （DHA）and 5-fluorouracil （5-FU）in combination on human gastric cancer cell line AGS in vitro .Methods Human gastric cancer line AGS was treated with different concentrations of DHA and 5-FU alone or in combination.The inhibition of cell proliferation was evaluated by MTT assay.Dose of median （IC50 ）of drugs （alone or in combination）and the combination index （CI）were calculated using the median-effect equation and the combination index equation of Chou-Talalay.Flow cytometry was used to detect the cell cycle distribution.The expression of mitochondrial respiratory membrane protein complex in AGS cells was analyzed with Western blot.Results DHA and 5-FU alone or in combination could markedly suppress the proliferation of AGS in significantly time-dependent and dose-dependent manners （P 〈0.05）.IC50 values with DHA or 5-FU administered for 24 h and 48 h were 5 1.60 μg/mL （DHA：24 h）,34.82 μg/mL （DHA：48 h）,45.90 μg/mL （5-FU：24 h）,and 1 6.86 μg/mL （5-FU：48 h）, respectively.DHA remarkably strengthened the inhibitory effect of 5-FU and decreased IC50 of 5-FU by 3.56 -2.1 5 folds.The combination of DHA and 5-FU showed synergism.Flow cytometry showed that AGS cells treated with DHA and 5-FU were arrested in G0/G1 phase and the proportion of AGS cells in G0/G1 phase increased compared with that in the control group,DHA group and 5-FU group,while the proportion of the cells in S phase decreased significantly （P 〈 0.05 ）.Western blot showed after treatment with DHA and 5-FU for 48 h,the expression of mitochondrial respiratory membrane protein complex was significantly decreased compared with control group,DHA group and 5-FU group （P 〈0.05）.Conclusion DHA could act synergistically with 5-FU in inhibiting the growth of gastric carcinoma cells,and meanwhile decrease the dose of 5-FU.The mechanism may be associated with cell cycle arrest in G0/G1 phase and interference in the energy metabolism of

关 键 词：二十二碳六烯酸 胃癌 5-氟尿嘧啶 线粒体呼吸链膜蛋白复合体 细胞周期 

分 类 号：R735.2[医药卫生—肿瘤]