低聚葡萄籽原花青素联合顺铂对A549细胞增殖及细胞周期的影响

Effect of Oligomeric Proanthocyanidins from Grape Seeds Combined with Cisplatin on A549 Cell Proliferation and Cell Cycle

目的:探讨低聚葡萄籽原花青素(oligomer grape seed proanthocyanidins,O-GSP)联合顺铂(cisdichlorodiamineplatinum(Ⅱ),DDP)对A549细胞增殖、细胞周期及细胞周期相关蛋白表达的影响。方法:体外培养A549细胞,四甲基偶氮唑蓝法检测DDP和O-GSP单独及联合用药对A549细胞存活率的影响;流式细胞仪检测O-GSP联合DDP对A549细胞周期的影响;Western Blot检测促细胞周期相关蛋白CDK 4、Cyclin D1和p-Rb的表达。结果:DDP(≥5 mg/L)和O-GSP(≥16 mg/L)单独用药均对A549细胞增殖有抑制作用,且两者同时作用对细胞增殖抑制具有联合作用。5 mg/L DDP单独用药可以于S期阻滞A549细胞,4 mg/L O-GSP单独用药可以于G_0/G_1期阻滞A549细胞,两者联合可显著降低细胞周期调控蛋白CDK 4、Cyclin D1和p-Rb的表达(P<0.05)。结论:O-GSP可以联合DDP抑制A549细胞增殖,且这种联合作用与Cyclin D1-CDK 4-Rb通路介导的细胞周期调控有关。

Objective:To investigate the effect of oligomeric proanthocyanidins from grape seeds(O-GSP)combined with cisplatin(DDP)on the proliferation,cell cycle and cell cycle-related protein expression of A549cells.Methods:A549cells were cultured in vitro.The effects of DDP and O-GSP alone and in combination on the survival rate of A549cells were observed by MTT assay;the combined effect of O-GSP and DDP on the cell cycle of A549cells was detected by flow cytometry and the expression levels of the cell cycle-related proteins CDK4,Cyclin D1and p-Rb were detected by Western blot.Results:DDP(≥5mg/L)and O-GSP(≥16mg/L)treatments alone could inhibit A549cell proliferation.The combined treatment had a synergistic effect on the inhibition of cell proliferation.DDP treatment alone at5mg/L arrested A549cells at S phase and O-GSP at4mg/L arrested the cells at G0/G1phase.The combination of DDP at5mg/L and O-GSP at4mg/L significantly reduced the expression levels of the cell cycle regulatory proteins CDK4,Cyclin D1and p-Rb.Conclusion:DDP can inhibit the proliferation of A549cells in combination with O-GSP through a mechanism related to the regulation of cell cycle progression mediated by the Cyclin D1-CDK4-Rb pathway.