南方医科大学学报 ›› 2015, Vol. 35 ›› Issue (08): 1103-.

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三七总皂苷对K562细胞增殖、凋亡及周期的影响及机制

李玉云,翟玮玮,杨向荣,丁娟,阚立新   

  • 出版日期:2015-08-20 发布日期:2015-08-20

Effects of Panax notoginseng saponins on proliferation, apoptosis and cell cycle of K562
cells in vitro and the mechanisms

  • Online:2015-08-20 Published:2015-08-20

摘要: 目的研究三七总皂苷(PNS)对K562 细胞增殖、凋亡、周期的影响及其相关分子机制。方法采用MTT法检测PNS对
K562细胞增殖的影响;AO/EB双荧光染色法及Annexin V-FITC/PI双染色法观察细胞凋亡及死亡状况;流式细胞术检测K562
细胞的周期变化;RT-PCR检测mTOR信号通路主要分子mRNA的表达变化;Western blot 检测cleaved caspase-3 蛋白、cyclin
D1蛋白及mTOR信号通路主要蛋白及磷酸化蛋白的表达量变化。结果100~800 μg/mL的PNS 作用于K562细胞后能够抑制
细胞增殖。PNS能够上调cleaved caspase-3蛋白表达,促使K562细胞发生凋亡。并且下调cyclin D1蛋白表达,促使K562细胞
阻滞在G0/G1期。同时,PNS 能够抑制K562 细胞mTOR mRNA 表达,降低mTOR 蛋白和磷酸化蛋白p-mTOR(Ser2448)、
p-p70S6K(Thr229/389)及p-4E-BP1(Thr37/46)的表达,从而抑制mTOR信号通路活性。结论PNS 对体外培养K562细胞有抑
制增殖,促进凋亡,使其发生周期阻滞的作用。其机制可能与PNS抑制mTOR信号通路活性、上调cleaved caspase-3蛋白表达
及抑制cyclin D1蛋白表达有关。

Abstract: Objective To investigate the effects of Panax notoginseng saponins (PNS) on the proliferation, apoptosis and cell cycle
of K562 cells and explore the molecular mechanisms underlying these effects. Methods PNS-induced growth inhibition of
K562 cells was detected by MTT assay; the cell apoptosis was evaluated by AO/EB staining and Annexin V-FITC/ PI staining;
flow cytometry was used to detect cell cycle changes in the treated cells. The mRNA expressions of the molecules in mTOR
signaling pathway were examined by RT-PCR, and the cellular expressions of cleaved caspeas-3, cyclin D1 and major
proteins in mTOR signaling pathway were detected using Western blotting. Results MTT assay showed that treatment with
100-800 μg/mL PNS significantly inhibited the proliferation, promoted the cell apoptosis, and caused cell cycle arrest in G0/G1
phase in K562 cells. Western blotting revealed increased protein expression of cleaved caspase-3 and decreased expression of
cyclin D1 in PNS-treated cells, in which the proteins expressions of mTOR, p-mTOR, p-p70S6K and p-4E-BP 1 and the mRNA
expression of mTOR were all decreased. Conclusion PNS can inhibit the proliferation, induce apoptosis and cause cell cycle
arrest in K562 cells possibly by up-regulating cleaved caspase 3 and down-regulating cyclin D1 and mTOR signaling pathway.