消痰散结方对人结直肠癌HCT116细胞增殖和凋亡及Caspase3蛋白表达的影响

Effect of Xiaotan Sanjie Recipe on Proliferation and Apoptosis of HCT116 Cell and Expression of Caspase3 Protein

目的探讨消痰散结方对结直肠癌细胞系增殖、凋亡的影响。方法消痰散结方浸膏冻干粉末溶解在细胞培养液中,制备消痰散结方保存液。以不同浓度消痰散结方作用HCT116细胞72 h,CCK-8法检测细胞增殖;取0.94、1.88 mg/m L消痰散结方作用于HCT116细胞24、48、72 h,CCK-8法检测细胞增殖;消痰散结方(0.24、0.47、0.94、1.88 mg/m L)作用细胞72 h后,TUNEL法原位检测细胞凋亡,计算细胞凋亡率;Western blot检测药物干预后Caspase3蛋白表达变化。结果消痰散结方对HCT116细胞增殖有抑制作用,作用72 h IC50为0.94 mg/m L,抑制作用呈浓度-时间依赖性,药物浓度和作用时间有交互作用;消痰散结方促进HCT116细胞凋亡,随药物浓度增高,凋亡率增高,与对照组比较差异有统计学意义(P<0.01);药物作用后Pro-Caspase3蛋白表达量降低,Cleaved Caspase3蛋白表达量增加。结论消痰散结方在体外对HCT116细胞有抑制增殖、促进凋亡的作用,其机制可能与增强Caspase3活性相关。

Objective To investigate the effects of Xiaotan Sanjie Recipe （XTSJR） on the proliferation and apoptosis of colorectal cancer cell;To study its relevant mechanism. Methods The original XTSJR aqueous solution was lyophilized, weighted, and dissolved in cell culture and stored. HCT116 cell line was treated with different concentrations of XTSJR for 72 h, and the cell proliferation was detected by CCK-8 assay. XTSJR treated HCT116 cell line with the concentration of 0.94 mg/mL and 1.88 mg/mL for 24 h, 48 h, 72 h, and the cell proliferation was detected by CCK-8 assay. HCT116 cell line was treated with different concentrations （0.24, 0.47, 0.94, 1.88 mg/mL） of XTSJR for 72 h. The cell apoptosis was detected by TUNEL assay under the fluorescence microscope, and the post-intervention expression of Caspase3 protein was detected by Western blot. Results The inhibitory effects of XTSJR on the proliferation of HCT116 cell line was concentration-time dependent. The IC50 value at 72 h-time point was 0.94 mg/mL. Medicine concentration and the treated time have interactive contribution to the inhibitory effect. XTSJR induced the cell apoptosis. The apoptosis rate increased with the increasing medicine concentration. There was statistically significant difference compared with the control group （ P〈0.01）. After treatment the expression of Pro-Caspase3 decreased, while the expression Cleaved Caspase3 protein increased. Conclusion XTSJR inhibits the proliferation of HCT116 cell line and induce apoptosis. Its mechanism might be related to the activity of Caspase3 protein.