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目的探讨大蒜素对人胃癌细胞系MGC803和SGC7901细胞周期的影响及其作用机制。方法以台盼蓝拒染法检测人胃癌细胞系MGC803和SGC7901细胞的增殖抑制率;以透射电镜观察两种胃癌细胞的直接损伤改变;以流式细胞仪及瑞士姬姆萨染色光镜观察两种胃癌细胞周期的改变;以SP免疫组化及RTPCR方法检测两种胃癌细胞的p21WAF1和p16INK4基因及蛋白表达的变化。结果大蒜素可抑制MGC803细胞生长,24h药物半数抑制浓度(IC50)为6.4μg/ml;也可抑制SGC7901细胞的生长,24hIC50为7.3μg/ml。12μg/ml的大蒜素作用两种胃癌细胞24h后,细胞出现急性直接损伤,膜系统改变明显。以3μg/ml、6μg/ml、9μg/ml终浓度大蒜素分别作用于两种胃癌细胞系24h后,与对照组比较,G0和G1期细胞周期百分比(%)明显减少,而G2和M期明显增加(P<0.01);6μg/ml大蒜素作用培养两种胃癌细胞24h后,与对照组比较,细胞分裂指数明显增加,提示两种细胞系经大蒜素处理后,细胞周期阻滞于M期。经大蒜素处理后,MGC803细胞的p21WAF1和p16INK4基因及蛋白表达均明显增加;SGC7901细胞的p21WAF1基因及蛋白表达明显增加。结论大蒜素可使MGC803及SGC7901两种细胞周期阻滞于M期;大蒜素的M期阻滞作用可能与其上调细胞的p21WAF1和p16INK4基因表达有关。
Objective To investigate the effects of allicin on the cell cycle of human gastric cancer cell lines MGC803 and SGC7901 and its mechanism. Methods The inhibitory rate of proliferation of human gastric cancer cell lines MGC803 and SGC7901 was detected by trypan blue exclusion staining. The direct damage of the two kinds of gastric cancer cells was observed by transmission electron microscopy. Flow cytometry and Giemsa staining of Swiss The changes of cell cycle were detected by immunohistochemistry and RTPCR method. The expressions of p21WAF1 and p16INK4 in two gastric cancer cell lines were detected by Western blot. Results Allicin inhibited the growth of MGC803 cells. The IC50 of 24h was 6.4μg / ml. Allicin also inhibited the growth of SGC7901 cells with 24hIC50 of 7.3μg / ml. 12μg / ml allicin effect of two kinds of gastric cancer cells 24h, the cells showed acute direct damage, the membrane system changes significantly. Allicin at 3μg / ml, 6μg / ml and 9μg / ml all showed a significant decrease in cell cycle percentage (%) at G0 and G1 phase compared with the control group, while G2 and M phase (P <0.01). After cultured with 6μg / ml Allicin for 24 h, the cell division index increased significantly compared with the control group, suggesting that the cell cycle arrest in both cell lines after allicin treatment was at M period. After allicin treatment, the gene and protein expression of p21WAF1 and p16INK4 in MGC803 cells were significantly increased; p21WAF1 gene and protein expression in SGC7901 cells increased significantly. Conclusion Allicin can arrest two kinds of cell cycle of MGC803 and SGC7901 in M phase. The arrest effect of allicin may be related to its up-regulation of p21WAF1 and p16INK4 gene expression.