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A novel polysaccharide from the Sarcodon aspratus triggers apoptosis in Hela cells via induction of mitochondrial dysfunction

机译: Sarcodon aspratus 的新型多糖通过诱导线粒体功能障碍来触发Hela细胞凋亡

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Background Polysaccharides extracted from fungus that have been used widely in the food and drugs industries due to biological activities. Objective The objective of the present study was to investigate the tumor-suppressive activity and mechanism of a novel polysaccharide (SAP) extracted from Sarcodon aspratus . Methods The SAP was extracted and purified using Sepharose CL-4B gel from S. aspratus . The cytotoxicity of SAP on cell lines was determined by MTT method. Cellular migration assays were implemented by using transwell plates. The apoptosis and mitochondrial membrane potential (Δψm) of Hela cells were analyzed by flow cytometry. The western blot was used to determine the protein expression of Hela cells. Results The results showed that SAP with a molecular weight of 9.01×10~(5)Da could significantly inhibit the growth of Hela cells in vitro . Three-dimensional cell culture (3D) and transwell assays showed that SAP restrained the multi-cellular spheroids growth and cell migration. Flow cytometry analysis revealed that SAP induced a loss of mitochondrial membrane potential (Δψm). Western blot assays indicated that SAP promoted the release of cytochrome c, increased Bax expression, down-regulated of Bcl-2 expression and activated of caspase-3 expression. Conclusion This study suggested that SAP induced Hela cells apoptosis via mitochondrial dysfunction that are critical in events of caspase apoptotic pathways. The anti-tumor (Hela cells) activity of SAP recommended that S. aspratus could be used as a powerful medicinal mushroom against cancer.
机译:背景技术从真菌中提取的多糖由于其生物活性已广泛用于食品和药物工业。目的本研究的目的是研究一种提取自Sarcodon aspratus的新型多糖(SAP)的抑瘤活性及其机制。方法采用Sepharose CL-4B凝胶提取纯化的SAP。用MTT法测定SAP对细胞系的细胞毒性。细胞迁移测定是通过使用transwell板进行的。流式细胞仪分析了Hela细胞的凋亡和线粒体膜电位(Δψm)。免疫印迹用于确定Hela细胞的蛋白质表达。结果结果表明,分子量为9.01×10〜(5)Da的SAP可以显着抑制Hela细胞的生长。三维细胞培养(3D)和Transwell分析表明,SAP抑制了多细胞球体的生长和细胞迁移。流式细胞仪分析表明,SAP引起线粒体膜电位(Δψm)的损失。 Western印迹分析表明,SAP促进了细胞色素c的释放,Bax表达的增加,Bcl-2表达的下调和caspase-3表达的激活。结论该研究表明SAP通过线粒体功能障碍诱导Hela细胞凋亡,这在caspase凋亡途径中起关键作用。 SAP的抗肿瘤(Hela细胞)活性建议将S. aspratus用作抗癌的有力药用蘑菇。

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