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目的探讨鳕鱼皮寡肽(CSO)对人胃癌细胞(SGC-7901)的增殖影响。方法用不同浓度CSO处理体外培养的SGC-7901细胞后,荧光显微镜下观察细胞4’6-二脒基-2-苯基吲哚(DAPI)染色后细胞形态变化,应用CCK-8实验检测细胞活力,免疫印迹法和流式细胞术检测细胞凋亡及细胞周期。结果 CSO对SGC-7901细胞增殖有明显的抑制作用,且并呈剂量、时间效应关系(P<0.05)。作用于SGC-7901胃癌细胞24 h、48 h和72 h的IC50分别是156.90 g/L、102.10 g/L、73.13 g/L。DAPI染色后发现,SGC-7901细胞随着CSO浓度增加可见大量的细胞核碎裂,荧光强烈。24 h细胞凋亡率检测显示,细胞随着浓度的增加凋亡率呈上升趋势,表明CSO对SGC-7901呈浓度效应关系。细胞周期观察发现,SGC-7901细胞G1期细胞数随着CSO浓度的增加而递增,而S/G2期细胞随着浓度的细胞递降。Caspase-3、Caspase-9随着CSO浓度的逐渐增高表达上调,Bcl-2表达下调。结论鳕鱼皮寡肽能抑制人胃癌细胞(SGC-7901)增殖,诱导凋亡,抑癌机制可能与Caspase-3、Caspase-9上调和Bcl-2下调相关。
Objective To investigate the effect of salmon oligopeptide (CSO) on the proliferation of human gastric cancer cells (SGC-7901). Methods SGC-7901 cells cultured in vitro were treated with different concentrations of CSO. The morphological changes of cells after 4’6-dimercapto-2-phenylindole (DAPI) staining were observed under fluorescence microscope. Cells were detected by CCK-8 assay. Viability, immunoblotting and flow cytometry were used to detect apoptosis and cell cycle. RESULTS: CSO significantly inhibited the proliferation of SGC-7901 cells in a dose- and time-dependent manner (P<0.05). The IC50 of SGC-7901 gastric cancer cells at 24 h, 48 h, and 72 h were 156.90 g/L, 102.10 g/L, and 73.13 g/L, respectively. After DAPI staining, it was found that a large number of nuclei were fragmented and the fluorescence was intense in the SGC-7901 cells as the concentration of CSO increased. The detection of cell apoptosis rate at 24 h showed that the cell apoptosis rate increased with the increase of the concentration, indicating that CSO had a concentration-dependent effect on SGC-7901. The cell cycle observation showed that the number of G1 phase cells in SGC-7901 cells increased with the increase of the concentration of CSO, while the cells in S/G2 phase decreased with the concentration of cells. The expression of Caspase-3 and Caspase-9 increased with the increase of CSO concentration, and the expression of Bcl-2 was down-regulated. Conclusion Squid oligopeptide can inhibit the proliferation of human gastric cancer cells (SGC-7901) and induce apoptosis. The mechanism of tumor suppressor may be related to the up-regulation of Caspase-3 and Caspase-9 and the down-regulation of Bcl-2.