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目的研究猪苓多糖(polyporus polysaccharide,PPS)抑制肺癌A549细胞增殖的作用及其机制。方法设对照组和PPS低、中、高剂量组,用MTT实验、流式细胞术检测PPS对细胞增殖的影响,用实时荧光定量PCR(q RT-PCR)法检测PPS对A549细胞Cyclin D1 m RNA表达水平及其稳定性的影响,用Western blotting检测PPS对A549细胞Cyclin D1蛋白、人抗原R(human antigen R,Hu R)蛋白表达及对Hu R蛋白胞浆及胞核内分布的影响。结果与对照组比较,中、高剂量的PPS作用后,A549细胞增殖明显受到抑制(P<0.05),Cyclin D1 m RNA稳定性降低(P<0.05),Cyclin D1 m RNA及蛋白表达减少(P<0.05),Hu R总蛋白没有明显变化,但胞浆Hu R蛋白表达下降(P<0.05),胞核Hu R蛋白表达升高(P<0.05)。结论 PPS可抑制A549细胞增殖,其作用机制可能与改变Hu R在细胞内定位,从而降低Cyclin D1 m RNA稳定性及Cyclin D1蛋白表达有关。
Objective To study the effect of polyporus polysaccharide (PPS) on the proliferation of lung cancer A549 cells and its mechanism. Methods The effects of PPS on cell proliferation were detected by MTT assay and flow cytometry in control and PPS low, medium and high dose groups. Cyclin D1 m of A549 cells was detected by qRT-PCR The expression of Cyclin D1 protein, human antigen R (Hu R) protein and the distribution of Hu R protein in the cytoplasm and nucleus of A549 cells were detected by Western blotting. Results Compared with the control group, the proliferation of A549 cells was significantly inhibited (P <0.05), the stability of Cyclin D1 mRNA was decreased (P <0.05), and the expression of Cyclin D1 mRNA and protein was decreased <0.05). There was no significant change in Hu R protein, but the expression of Hu R protein in cytoplasm decreased (P <0.05) and the expression of Hu R protein in nucleus increased (P <0.05). Conclusions PPS can inhibit the proliferation of A549 cells and its mechanism may be related to the change of Hu R localization in cells and the decrease of Cyclin D1 m RNA stability and Cyclin D1 protein expression.