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目的:探讨仙慈丹(XCD)有效部位对肺癌细胞A549增殖抑制作用及其作用机制。方法:XCD有效部位为XCD水提滤液通过AB-8型大孔树脂柱收集80%乙醇洗脱液所得,得率为0.90%。体外培养A549细胞至对数期,分别以质量浓度为5,10,15,25,50,100,200 mg.L-1的仙慈丹有效部位作用,应用MTT法测定其对A549细胞的抑制率,进而求出半数抑制浓度(IC50)。运用AnnexinⅤ-FITC/PI双染流式细胞术及PI单染流式细胞术测定XCD有效部位对A549细胞凋亡及周期的影响。结果:通过MTT实验,经过统计计算得出XCD有效部位在对A549作用24,48,72 h后的IC50分别为136.550,56.671,55.322 mg.L-1;通过流式细胞仪测定XCD作用下A549在24,48,72 h的凋亡率分别为(17.98±0.11)%,(23.34±0.23)%,(29.54±0.78)%;XCD作用A549细胞24 h后,细胞周期被阻滞在S期,在48 h后细胞周期被阻滞在G0/G1期。结论:XCD有效部位对A549细胞增殖抑制作用明显,通过诱导肿瘤细胞凋亡,同时调节细胞周期抑制A549细胞生长。
Objective: To investigate the inhibitory effect of xidzidan (XCD) on the proliferation of lung cancer cell line A549 and its mechanism. Methods: The effective fraction of XCD was obtained by collecting the eluate of XCD water extract through 80% ethanol eluent through AB-8 macroporous resin column with a yield of 0.90%. A549 cells were cultured in vitro until the logarithmic phase, respectively, the concentration of 5,10,15,25,50,100,200 mg.L-1 cefoxitid effective site role, using MTT assay inhibition rate of A549 cells, and then seeking Half the inhibitory concentration (IC50). The effect of effective site of XCD on apoptosis and cycle of A549 cells was determined by Annexin V-FITC / PI double staining flow cytometry and PI single staining flow cytometry. Results: Through MTT experiments, the IC50 of effective fraction of XCD after 24,48,72 h of treatment with A549 were 136.550, 56.671 and 55.322 mg.L-1, respectively. The cell viability of A549 The apoptotic rates at 24,48,72 h were (17.98 ± 0.11)%, (23.34 ± 0.23)% and (29.54 ± 0.78)%, respectively. After XCD treated A549 cells for 24 h, the cell cycle was arrested in S phase After 48 h, the cell cycle was arrested at G0 / G1 phase. Conclusion: The effective site of XCD can inhibit the proliferation of A549 cells obviously, and induce the apoptosis of A549 cells by regulating the cell cycle and inhibiting the growth of A549 cells.