葡萄柚黄酮抗MDS细胞株Skm-1机制研究

Research on Mechanism of the Antagonistic Action ofPure Total Flavonoids from Citrus Paradisi Macfad on MDS Cell Line Skm-1

目的:观察葡萄柚黄酮(PTFC)对骨髓增生异常综合征(MDS)细胞株Skm-1的毒性,并探讨作用机制。方法:采用四氮唑盐法(MTT法)观察PTFC对MDS细胞株增殖抑制作用,用流式细胞术检测PTFC对细胞凋亡的诱导作用和对周期的干预作用,用Western blot检测相关蛋白表达。结果:PTFC呈时间-剂量依赖性抑制Skm-1细胞,24h、48h的IC50分别为1.54mg/mL,1.36mg/mL。流式细胞术检测凋亡结果显示,细胞早期凋亡率随着药物作用无明显增加,PTFC可阻滞细胞于G0/G1期。Western blotting结果显示PTFC未能激活Caspase-3、PARP,能引起CDK4、CDK6、CyclinD1、PI3K、Akt、P-Akt、P65下调。结论:PTFC对MDS细胞株有明显的毒性作用,其作用机制是通过PI3K/Akt/NF-kB信号通路使得细胞周期阻滞在G0/G1期,具有潜在的药物应用价值,值得进一步深入研究。

Objective To observe the effects of pure total flavonoids from Citrus paradisi Macfad（PTFC）on the proliferation in myelodysplastic syndrome（MDS） cell line Skm-1,and to investigate its mechanisms.Methods MTT assay was used to evaluate cell growth inhibition.Apoptosis and cell cycle were detected byflow cytometry analysis.The expression of proteins which were associated with apoptosis,cell cycle and PI3K/Akt/NF-kB signal pathway were examined by Western blotting.Results PTFC could inhibit the growth ofSkm-1 cells in a time-dose-dependent mode,with IC50 values of 1.54mg/mL for 24h and 1.36mg/mL for 48h.The results of flow cytometry analysis showed that cell apoptotic didn＇t increase following drug dose increasing,and the PTFC treated Skm-1 cells were blocked at the cell cycle arrest in G0/G1 phase.Western blotting resultsshowed that the expression of Caspase-3 and PARP were no changed,but the expressions of CDK4/CDK6,CyclinD1,PI3K,Akt,P-Akt and P65 were down-regulated after Skm-1 cells treated with PTFC.Conclusion PTFC hasobvious toxic effects on MDS cell lines,its mechanism might be making cell cycle be blocked in G0/G1 phase.Through PI3K/Akt/NF-kB signaling path ways PTFC has potential drug application value,is worthy of furtherlucubrated study.