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目的:观察健脾清热活血方对人结肠癌SW480细胞凋亡、增殖、周期及P-β-catenin蛋白的影响,探讨该方防治结肠癌可能机制。方法:培养人结肠癌SW480细胞,分别予空白胎牛血清、不同浓度健脾清热活血方含药血清及PI3K阻断剂LY294002干预24 h,MTT法及流式细胞术检测细胞增殖、凋亡及细胞周期,免疫荧光染色法检测P-β-catenin核内转位。结果:健脾清热活血方含药血清组细胞增殖抑制率及凋亡率显著高于空白对照组(P<0.05);同时S期细胞比例显著升高、G1期细胞比例显著降低(P<0.05)。健脾清热活血方含药血清组及LY294002组细胞P-β-catenin以膜表达为主,而空白对照组P-β-catenin以膜表达缺失及核内表达增加为主。结论:健脾清热活血方可能通过调控P-β-catenin核内转录,阻滞SW480细胞周期于G1期,诱导细胞凋亡,抑制细胞增殖,从而发挥防治结肠癌的效用。
Objective: To observe the effect of “Jianpi Qingre Huoxue Decoction” on the apoptosis, proliferation, cycle and P-β-catenin protein in human colon cancer SW480 cells and to explore the possible mechanism of this recipe in preventing and treating colon cancer. Methods: Human colorectal cancer SW480 cells were cultured and transfected with blank fetal bovine serum, different concentrations of drug-containing serum of Jianpi Qingre Huoxue Decoction and PI3K blocker LY294002 for 24 h. MTT assay and flow cytometry were used to detect cell proliferation and apoptosis Cell cycle, immunofluorescence staining P-β-catenin nuclear translocation. Results: The cell proliferation inhibition rate and apoptosis rate of the drug-containing serum of Jianpi Qingre Huoxue Recipe group were significantly higher than that of the blank control group (P <0.05). Meanwhile, the proportion of S phase cells was significantly increased and the proportion of cells in G1 phase was significantly decreased (P <0.05 ). The expression of P-β-catenin in Jianpi Qingre Huoxue Recipe containing LY294002 group and the drug-containing serum group of Jianpi Qingre Huoxue Recipe were the main, while the expression of P-β-catenin in the blank control group was mainly decreased. Conclusion: Jianpi Qingre Huoxue Recipe can exert the effect of preventing and treating colon cancer by regulating the nuclear transcription of P-β-catenin, arresting the cell cycle of SW480 in G1 phase, inducing cell apoptosis and inhibiting cell proliferation.