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Synergistic Apoptotic Effect of D-Fraction From Grifola frondosa and Vitamin C on Hepatocellular Carcinoma SMMC-7721 Cells

机译:灰树花D成分和维生素C对肝癌细胞SMMC-7721细胞的协同凋亡作用

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摘要

The aim of this study was to investigate the anticancer effect of a combination of D-fraction polysaccharide from Grifola frondosa (DFP) and vitamin C (VC) on hepatocellular carcinoma in vitro. DFP is a bioactive extract from the maitake mushroom. Anticancer activity was demonstrated using various concentrations of DFP alone or in combination with VC against the human hepatocarcinoma SMMC-7721 cell line. To investigate the anticancer mechanism, studies designed to detect cell apoptosis were conducted. Results from the MTT assay indicated that a combination of DFP (0.2 mg/mL) and VC (0.3 mmol/L) led to a 70% reduction in cell viability. Flow cytometry results indicated that DFP/VC treatment induced apoptosis in approximately 65% SMMC-7721 cells. Cell cycle analysis identified cell cycle arrest at the G2/M phase following DFP/VC treatment for 48 hours. In addition, cellular morphological changes were observed using transmission electron microscopy. Western blot analysis revealed that the upregulation of BAX, downregulation of Bcl-2, activation of poly-(ADP-ribose)-polymerase (PARP), and the release of cytochrome c were observed in cells treated with the combination of DFP/VC, which showed that the mechanism of anticancer activity in the SMMC-7721 hepatocarcinoma cells involved induction of apoptosis.
机译:这项研究的目的是研究Grifola frondosa(DFP)和维生素C(VC)的D级分多糖在体外对肝癌的抗癌作用。 DFP是舞茸的一种生物活性提取物。使用各种浓度的DFP单独或与VC结合使用,可证明其对人肝癌SMMC-7721细胞系具有抗癌活性。为了研究抗癌机制,进行了旨在检测​​细胞凋亡的研究。 MTT分析的结果表明,DFP(0.2 mg / mL)和VC(0.3 mmol / L)的组合导致细胞活力降低70%。流式细胞术结果表明,DFP / VC处理可诱导约65%的SMMC-7721细胞凋亡。细胞周期分析确定了DFP / VC处理48小时后G2 / M期的细胞周期停滞。另外,使用透射电子显微镜观察到细胞形态变化。 Western blot分析显示,在DFP / VC组合处理的细胞中,观察到BAX的上调,Bcl-2的下调,聚(ADP-核糖)-聚合酶(PARP)的激活以及细胞色素c的释放,这表明SMMC-7721肝癌细胞的抗癌活性机制涉及凋亡的诱导。

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