Objective To investigate the effect of water extracts of Coptidis Rhizoma and Evodiae Fructus at different ratios on the proliferation,migration and apoptosis of colon cancer cell CT-26 and to determine the optimal proportion of both. Methods Using MTT assay,clone formation assay,the wound healing and flow cytometry the proliferation,migration and apoptosis ability of CT-26 cells in experimental groups (Coptidis Rhizoma:Evodiae Fructus ratios at 1:6、2:5、3:4、4:3、5:2、6:1 and single of the two medicines) were detected. Results Coptidis Rhizoma,5:2,6:1 groups showed a remarkable inhibition effect on CT-26 ability of proliferation. After a 6h treatment of 6:1 group,there was an obvious inhibition of cell growth and it was significantly higher than that of 5:2 and Coptidis Rhizoma group (P<0.01). After 48h treatment of 6:1,5:2 and optidis Rhizoma groups,the proliferation inhibition rates reached (80.39±2.72)%,(56.34±12.57)% and (62.71±5.46)%,respectively. In addition,the growth inhibition effect exhibited a time-dependent and dose-dependent pattern. As the proportion of Coptidis Rhizoma in experiment groups increased,the clone formation rate decreased gradually and in 2:5,3:4,4:3,5:2,6:1 and Coptidis Rhizoma groups the rates were lower significantly than that in the control group (P<0.01). The wound healing indicated that Coptidis Rhizoma and 6:1 groups' 48h treatment could obviously inhibite the wound healing process. Flow cytometry analysis showed that with the increase of Coptidis Rhizoma the apoptosis rates increased and after 24h treatment of Coptidis Rhizoma and 6:1 groups the rates were (31.05± 1.34)%, (35.09 ±3.72)%(P<0.05). Conclusion The combination of Coptidis Rhizoma and Evodiae Fructus can inhibit the proliferation,migration and induce apoptosis of CT-26 cells,and the strongest effect is achieved at the proportion of Coptidis Rhizoma and vodiae Fructus at 6:1.%目的 比较黄连、吴茱萸不同配比的水提物对结肠癌细胞CT-26增殖、迁移和凋亡的影响,从而找出两药的最佳配比.方法 采用MTT试验、克隆形成试验、细胞划痕试验、流式细胞术观察不同配比(黄连:吴茱萸比例为1:6、2:5、3:4、4:3、5:2、6:1以及两药单独作用)的水提物对CT-26细胞增殖、迁移和凋亡的影响. 结果 黄连、5:2、6:1组对细胞增殖的抑制作用明显,6:1组作用6 h后即产生抑制作用,12、24、48 h的细胞增殖抑制率均大于5:2组和黄连组,且差异有统计学意义(P<0.01),6:1、5:2 和黄连组作用48 h后的增殖抑制率分别达到(80.39± 2.72)%、(56.34±12.57)%、(62.71±5.46)%,且黄连、吴茱萸配伍对细胞增殖的抑制作用存在时间依赖性和剂量依赖性. 随着配比中黄连比例的增加,克隆形成率逐渐降低,2:5、3:4、4:3、5:2、6:1、黄连组的集落形成率与空白对照组相比差异均有统计学意义(P<0.01). 划痕实验结果表明黄连组和6:1组作用24 h后,与空白对照组相比能够明显抑制划痕愈合的程度.流式细胞仪PI单染亚二倍体峰分析结果显示,随着配比中黄连比例的增加,细胞凋亡率上升,作用24 h后黄连和6:1 组的细胞凋亡率分别达到(31.05±1.34)%、(35.09±3.72)%,且两者差异有统计学意义(P<0.05). 结论 黄连、吴茱萸配伍对结肠癌细胞CT-26的增殖、迁移和凋亡均有一定的抑制作用,其中以配比为6:1时作用最显著.
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