二十二碳六烯酸与5-氟尿嘧啶通过干扰人胃癌AGS细胞系的能量代谢和细胞周期阻滞发挥协同抗癌特性。
Synergistic anticancer properties of docosahexaenoic acid and 5-fluorouracil through interference with energy metabolism and cell cycle arrest in human gastric cancer cell line AGS cells.
作者信息
Gao Kun, Liang Qi, Zhao Zhi-Hao, Li You-Fen, Wang Shu-Feng
机构信息
Kun Gao, You-Fen Li, The Key Laboratory of Biomedical Information Engineering of Ministry of Education, School of Life Science and Technology, Xi'an Jiaotong University, Xi'an 710049, Shaanxi Province, China.
出版信息
World J Gastroenterol. 2016 Mar 14;22(10):2971-80. doi: 10.3748/wjg.v22.i10.2971.
AIM
To explore the synergistic effect of docosahexaenoic acid (DHA)/5-fluorouracil (5-FU) on the human gastric cancer cell line AGS and examine the underlying mechanism.
METHODS
AGS cells were cultured and treated with a series of concentrations of DHA and 5-FU alone or in combination for 24 and 48 h. To investigate the synergistic effect of DHA and 5-FU on AGS cells, the inhibition of cell proliferation was determined by MTT assay and cell morphology. Flow cytometric analysis was also used to assess cell cycle distribution, and the expression of mitochondrial electron transfer chain complexes (METCs) I, II and V in AGS cells was further determined by Western blot analysis.
RESULTS
DHA and 5-FU alone or in combination could markedly suppress the proliferation of AGS cells in a significant time and dose-dependent manner. DHA markedly strengthened the antiproliferative effect of 5-FU, decreasing the IC50 by 3.56-2.15-fold in an apparent synergy. The morphological changes of the cells were characterized by shrinkage, cell membrane blebbing and decreased adherence. Cell cycle analysis showed a shift of cells into the G0/G1 phase from the S phase following treatment with DHA or 5-FU (G0/G1 phase: 30.04% ± 1.54% vs 49.05% ± 6.41% and 63.39% ± 6.83%, respectively, P < 0.05; S phase: 56.76% ± 3.14% vs 34.75% ± 2.35% and 25.63% ± 2.21%, respectively, P < 0.05). Combination treatment of DHA and 5-FU resulted in a significantly larger shift toward the G0/G1 phase and subsequent reduction in S phase (G0/G1 phase: 69.06% ± 2.63% vs 49.05% ± 6.41% and 63.39% ± 6.83%, respectively, P < 0.05; S phase: 19.80% ± 4.30% vs 34.75% ± 2.35% and 25.63% ± 2.21%, respectively, P < 0.05). This synergy was also reflected in the significant downregulation of the expression of METCs in AGS cells.
CONCLUSION
Synergistic anticancer properties of DHA and 5-FU may involve interference with energy production of AGS cells via downregulation of METCs and cell cycle arrest.
目的
探讨二十二碳六烯酸（DHA）/5-氟尿嘧啶（5-FU）对人胃癌细胞系AGS的协同作用，并研究其潜在机制。
方法
培养AGS细胞，分别用一系列浓度的DHA和5-FU单独或联合处理24小时和48小时。为研究DHA和5-FU对AGS细胞的协同作用，通过MTT法和细胞形态学检测细胞增殖抑制情况。还采用流式细胞术分析评估细胞周期分布，并通过蛋白质免疫印迹分析进一步测定AGS细胞中线粒体电子传递链复合物（METCs）Ⅰ、Ⅱ和Ⅴ的表达。
结果
DHA和5-FU单独或联合使用均可显著抑制AGS细胞增殖，呈明显的时间和剂量依赖性。DHA显著增强了5-FU的抗增殖作用，使半数抑制浓度（IC50）明显协同降低3.56至2.15倍。细胞形态学变化表现为细胞皱缩、细胞膜起泡和贴壁减少。细胞周期分析显示，用DHA或5-FU处理后，细胞从S期向G0/G1期转变（G0/G1期：分别为30.04%±1.54%、49.05%±6.41%和63.39%±6.83%，P<0.05；S期：分别为56.76%±3.14%、34.75%±2.35%和25.63%±2.21%，P<0.05）。DHA和5-FU联合处理导致向G0/G1期的转变明显更大，随后S期减少（G0/G1期：分别为69.06%±2.63%、49.05%±6.41%和63.39%±6.83%，P<0.05；S期：分别为19.80%±4.30%、34.75%±2.35%和25.63%±2.21%，P<0.05）。这种协同作用还体现在AGS细胞中METCs表达的显著下调。
结论
DHA和5-FU的协同抗癌特性可能涉及通过下调METCs干扰AGS细胞的能量产生和细胞周期阻滞。
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