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首页> 外文期刊>World Journal of Gastroenterology >Molecular mechanisms of apoptosis induced by Scorpio water extract in human hepatoma HepG2 cells.
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Molecular mechanisms of apoptosis induced by Scorpio water extract in human hepatoma HepG2 cells.

机译：天蝎水提取物诱导人肝癌HepG2细胞凋亡的分子机制。

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AIM: To clarify the mechanism underlying the anti-mutagenic and anti-cancer activities of Scorpio water extract (SWE). METHODS: Human hepatoma HepG2 cells were incubated with various concentrations of SWE. After 24-h incubation, cytotoxicity and apoptosis evaluations were determined by MTT and DNA fragmentation assay, respectively. After treatment with SWE, mitochondrial membrane potential (MMP) was determined by measuring the retention of the dye 3,3'-dihexyloxacarbocyanine (DiOC(6)(3)) and the protein expression including cytochrome C and poly-(ADP-ribose) polymerase (PARP) were measured by Western blotting. Caspase-3 and -9 enzyme activities were measured using specific fluorescence dyes such as Ac-DEVD-AFC and Ac-LEHD-AFC. RESULTS: We found that treatment with SWE induced apoptosis as confirmed by discontinuous DNA fragmentation in cultured human hepatoma HepG2 cells. Our investigation also showed that SWE-induced apoptosis of HepG2 cells were associated with intracellular events including disruption of MMP, increased translocation of cytochrome C from mitochondria to cytosol, activation of caspase-3, and PARP. Pre-treatment of N-acetyl-Asp-Glu-Val-Asp-CHO (Ac-DEVD-CHO), a caspase-3 specific inhibitor, or cyclosporin A (CsA), an inhibitor of MMP disruption, completely abolished SWE-induced DNA fragmentation. CONCLUSION: These results suggest that SWE possibly causes mitochondrial damage, leading to cytochrome C release into cytosol and activation of caspases resulting in PARP cleavage and execution of apoptotic cell death in HepG2 cells. These results further suggest that Scorpio may be a valuable agent of therapeutic intervention of human hepatomas.
机译：目的：阐明天蝎座水提取物（SWE）的抗诱变和抗癌活性的机制。方法：将人肝癌HepG2细胞与各种浓度的SWE孵育。孵育24小时后，分别通过MTT和DNA片段测定法确定细胞毒性和凋亡评估。用SWE处理后，通过测量染料3,3'-二己基氧杂羰花青素（DiOC（6）（3））的保留和蛋白质表达（包括细胞色素C和聚-（ADP-核糖））来确定线粒体膜电位（MMP）通过蛋白质印迹法测量聚合酶（PARP）。使用特定的荧光染料（例如Ac-DEVD-AFC和Ac-LEHD-AFC）测量Caspase-3和-9酶的活性。结果：我们发现用SWE处理可诱导凋亡，这一点已通过培养的人肝癌HepG2细胞中不连续的DNA片段得到证实。我们的研究还显示，SWE诱导的HepG2细胞凋亡与细胞内事件有关，包括MMP破坏，细胞色素C从线粒体到细胞质的易位性增加，胱天蛋白酶3的活化和PARP。预处理caspase-3特异性抑制剂N-乙酰-Asp-Glu-Val-Asp-CHO（Ac-DEVD-CHO）或MMP破坏抑制剂环孢菌素A（CsA）完全废除了SWE诱导的DNA片段化。结论：这些结果表明，SWE可能引起线粒体损伤，导致细胞色素C释放到细胞质中并激活半胱氨酸蛋白酶，导致PARP裂解并导致HepG2细胞凋亡。这些结果进一步表明，天蝎座可能是治疗人类肝癌的有价值的药物。

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