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Cisplatin combination drugs induce autophagy in HeLa cells and interact with HSA via electrostatic binding affinity

机译:顺铂联合药物诱导HeLa细胞自噬并通过静电结合亲和力与HSA相互作用

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Cisplatin, as a significant chemotherapeutic drug for the treatment of cancers, was combined with rapamycin (RAPA), an autophagy inducer, or 3-methyladenine (3-MA), an autophagy inhibitor, and these cisplatin combination drugs were tested with HeLa cells to explore their specific effects on autophagy by cell viability assay, mitochondria membrane potential (MMP) determination, transmission electron microscopic (TEM) observation, dansylcadaverine (MDC) staining, and western blotting analysis. Results revealed that cisplatin combination drugs enhanced formation of autophagosomes, and morphological and biochemical markers of autophagy in HeLa cells can be clearly determined with the formation of enlarged acidic vesicles and conversion of light chain 3 (LC3) protein. Cisplatin combination drugs induce stronger effects on autophagy than either of the components does. Combination drug-induced autophagy inhibits the growth of HeLa cell in a dose-dependent manner and subsequently sensitizes the cells to apoptosis and cell death. Furthermore, interactions between cisplatin combination drugs and human serum albumin (HSA) were investigated under fluorescence, synchronous fluorescence, and circular dichroism analysis. Results suggest that cisplatin combination drugs can bind to HSA and induce conformation and microenvironmental changes of HSA via electrostatic binding affinity. These investigations can provide useful and fundamental information, which could be used in cytotoxic chemotherapy to dramatically increase efficacy in pharmaceutical and biotechnology fields.
机译:顺铂是治疗癌症的重要化学治疗药物,与自噬诱导剂雷帕霉素(RAPA)或自噬抑制剂3-甲基腺嘌呤(3-MA)结合使用,并用HeLa细胞对这些顺铂组合药物进行了测试,通过细胞活力测定,线粒体膜电位(MMP)测定,透射电镜(TEM)观察,丹磺胺(MDC)染色和蛋白质印迹分析探索它们对自噬的特异性作用。结果表明,顺铂联合药物可增强自噬体的形成,并且通过扩大酸性囊泡的形成和轻链3(LC3)蛋白质的转化,可以清楚地确定HeLa细胞中自噬的形态学和生化标志物。顺铂联合药物对自噬的诱导作用强于任一组分。组合药物诱导的自噬以剂量依赖的方式抑制HeLa细胞的生长,并随后使细胞对凋亡和细胞死亡敏感。此外,在荧光,同步荧光和圆二色性分析下,研究了顺铂联合药物与人血清白蛋白(HSA)之间的相互作用。结果表明顺铂联合药物可以通过静电结合亲和力与HSA结合并诱导HSA 的构象和微环境变化。这些研究可以提供有用的基础信息,这些信息可用于细胞毒性化学疗法,以大大提高药物和生物技术领域的功效。

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