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目的:探讨健脾活血解毒法拆方含药血清对人结直肠癌细胞系HCT-116细胞的生长、细胞凋亡率、细胞凋亡相关基因和自噬基因表达的影响。方法:实验分为对照组、健脾组、活血组、解毒组、健脾活血组、健脾解毒组和健脾活血解毒组,分别采用酸性磷酸酶法(APA)、流式细胞术(FCM)和Western blot法检测各组含药血清对肿瘤细胞的生长、细胞凋亡率的变化、凋亡相关基因Caspase-3表达以及自噬相关基因LC3表达。结果:各组10%含药血清作用24 h后均能抑制HCT-116细胞增殖,但组间不具有明显差异。光镜下可见,健脾活血解毒法各组含药血清作用24 h后,细胞数量减少,且体积变小;细胞早期及中晚期凋亡率与对照组相比无显著性差异,但自噬相关蛋白LC3出现活性剪切条带。结论:健脾活血解毒法含药各拆方血清体外可抑制人结直肠癌HCT-116细胞增殖,机制可能与自噬作用通路密切相关。
Objective: To investigate the effect of Jianpi Huoxue Jiedu Drug-containing Serum on the growth, apoptosis rate, apoptosis-related genes and autophagy gene expression of human colorectal cancer cell line HCT-116. Methods: The experiment was divided into the control group, the spleen group, the Huoxue group, the detoxification group, the Jianpihuoxue group, the Jianpijiedu group and the Jianpi Huoxueetuetu group, respectively. The cells were treated with acid phosphatase (APA), flow cytometry ) And Western blot were used to detect the changes of tumor cell growth and apoptosis rate, the expression of Caspase-3, and the expression of autophagy-related gene LC3 in each group. Results: The proliferation of HCT-116 cells was inhibited by 10% drug-containing serum in each group for 24 h, but there was no significant difference between the two groups. Light microscope, the spleen promoting blood circulation and detoxification method in each group containing serum 24 h, the number of cells decreased, and smaller; early and late apoptosis rate compared with the control group no significant difference, but the autophagy Associated protein LC3 active shear band. Conclusion: The spleen and promoting blood circulation to detoxification drugs containing the demolition of serum in vitro can inhibit the proliferation of human colorectal cancer HCT-116 cells, the mechanism may be closely related with autophagy pathway.