The induction of apoptosis and autophagy in human hepatoma SMMC-7721 cells by combined treatment with vitamin C and polysaccharides extracted from Grifola frondosa
The induction of apoptosis and autophagy in human hepatoma SMMC-7721 cells by combined treatment with vitamin C and polysaccharides extracted from Grifola frondosa
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DOI：
10.1007/s10495-017-1421-z
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发表时间：
2017-11-01
期刊：
影响因子：
7.2
通讯作者：
Yang, Ke-Hu
中科院分区：
文献类型：
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作者：
Zhao, Fei;Zhao, Jin;Yang, Ke-Hu
从蘑菇 Grifola frondosa (GFP) 中提取的多糖是一种潜在的抗癌剂。本研究的目的是研究单独使用 GFP 和维生素 C (VC) 以及联合使用对人肝癌 SMMC-7721 和 HepG2 细胞活力的影响。还进行了旨在检测细胞凋亡和自噬的研究来探讨其机制。细胞活力测定结果表明，24小时后，GFP（0.2或0.25 mg/mL）和VC（0.3 mmol/L）（GFP/VC）组合导致SMMC-7721和HepG2细胞的细胞活力分别降低52.73%和53.93%。流式细胞分析表明，GFP/VC处理诱导细胞周期停滞在G2/M期，SMMC-7721和HepG2细胞中分别有约43.62%和42.46%发生细胞凋亡。此外，Hoechst33258和单丹磺酰尸胺染色以及透射电子显微镜的结果表明，GFP/VC诱导SMMC-7721和HepG2细胞的凋亡和自噬。蛋白质印迹分析显示凋亡相关蛋白[BAX和caspase-3上调、Bcl-2下调和聚（ADP-核糖）聚合酶激活]和自噬蛋白标记物（beclin-1和微管相关蛋白1A/1B轻链-3上调）表达的变化。我们还证明 Akt 和 p-Akt 的表达均增强，表明 PI3K/Akt/mTOR 通路可能不参与该过程。我们的研究表明，GFP和VC的联合应用在体外诱导细胞凋亡和自噬，并可能在体内具有抗肿瘤活性。
Polysaccharides extracted from the mushroom Grifola frondosa (GFP) are a potential anticancer agent. The objective of this study was to investigate the effect of GFP and vitamin C (VC) alone and in combination on the viability of human hepatocarcinoma SMMC-7721 and HepG2 cells. Studies designed to detect cell apoptosis and autophagy were also conducted to investigate the mechanism. Results from the cell viability assay indicated that a combination of GFP (0.2 or 0.25 mg/mL) and VC (0.3 mmol/L) (GFP/VC) led to 52.73 and 53.93% reduction in cell viability of SMMC-7721 and HepG2 cells separately after 24 h. Flow cytometric analysis indicated that GFP/VC treatment induced cell cycle arrest at the G2/M phase, and apoptosis occurred in approximately 43.62 and 42.46% of the SMMC-7721 and HepG2 cells separately. Moreover, results of Hoechst33258 and monodansylcadaverine staining, and transmission electron microscopy, showed that GFP/VC induced apoptosis and autophagy in SMMC-7721 and HepG2 cells. Western blot analysis showed changes in the expression of apoptosis-related proteins [upregulation of BAX and caspase-3, downregulation of Bcl-2, and activation of poly-(ADP-ribose)-polymerase] and autophagy protein markers (upregulation of beclin-1 and microtubule-associated protein 1A/1B light chain-3). We also demonstrated that the expression of both Akt and p-Akt was enhanced, suggesting the PI3K/Akt/mTOR pathway might not be involved in this process. Our study shows that the combined application of GFP and VC induced cell apoptosis and autophagy in vitro, and might have antitumor activity in vivo.