首页> 外文期刊>Cancer Cell International >Selected polyphenols potentiate the apoptotic efficacy of glycolytic inhibitors in human acute myeloid leukemia cell lines. Regulation by protein kinase activities
【24h】

Selected polyphenols potentiate the apoptotic efficacy of glycolytic inhibitors in human acute myeloid leukemia cell lines. Regulation by protein kinase activities

机译:选择的多酚可增强糖酵解抑制剂在人急性髓性白血病细胞系中的凋亡功效。通过蛋白激酶活性调节

获取原文
获取外文期刊封面目录资料

摘要

Background The glycolysis inhibitor 2-deoxy- d -glucose (2-DG) is a safe, potentially useful anti-tumour drug, but its efficacy is normally low when used alone. Recent studies indicated that 2-DG stimulates the PI3K/Akt and MEK/ERK defensive pathways, which limits the apoptotic efficacy in tumour cell lines. We hypothesized that co-treatment with selected polyphenols could improve 2-DG-provoked apoptosis by preventing defensive kinase activation. Methods Cell proliferation was measured by cell counting or the MTT assay. Cell cycle, apoptosis and necrosis were determined by propidium iodide staining and/or annexin V labeling followed by flow cytometry. Mitochondria pore transition and depolarization were determined by calcein-ATM or rhodamine 123 labeling followed flow cytometry. Intracellular reactive oxygen species and GSH were determined by dichlorodihydrofluorescein diacetate or monochlorobimane labeling followed by flow cytometry or fluorimetry. Expression and phosphorylation of protein kinases were analyzed by the Western blot. Results (i) 2-DG-provoked apoptosis was greatly potentiated by co-treatment with the sub-lethal concentrations of the flavonoid quercetin in human HL60 acute myeloblastic leukemia cells. Allowing for quantitative differences, apoptosis potentiation was also obtained using NB4 promyelocytic and THP-1 promonocytic cells, using curcumin or genistein instead of quercetin, and using lonidamine instead of 2-DG, but not when 2-DG was substituted by incubation in glucose-free medium. (ii) Quercetin and 2-DG rapidly elicited the opening of mitochondria pore transition, which preceded the trigger of apoptosis. (iii) Treatments did not affect GSH levels, and caused disparate effects on reactive oxygen species generation, which did not match the changes in lethality. (iv) 2-DG and lonidamine stimulated defensive Akt and ERK phosphorylation/activation, while glucose starvation was ineffective. Polyphenols prevented the stimulation of Akt phosphorylation, and in some cases also ERK phosphorylation. In addition, quercetin and 2-DG stimulated GSK-3α,β phosphorylation/inactivation, although with different isoform specificity. The use of pharmacologic inhibitors confirmed the importance of these kinase modifications for apoptosis. Conclusions The present in vitro observations suggest that co-treatment with low concentrations of selected polyphenols might represent a manner of improving the poor anti-tumour efficacy of some glycolytic inhibitors, and that apoptosis potentiation may be at least in part explained by the regulation of defensive protein kinase activities.
机译:背景技术糖酵解抑制剂2-deoxy-d-glucose(2-DG)是一种安全,潜在有用的抗肿瘤药物,但单独使用时其功效通常较低。最近的研究表明2-DG刺激PI3K / Akt和MEK / ERK防御途径,这限制了肿瘤细胞系的凋亡效力。我们假设与选定的多酚类共同治疗可通过预防防御性激酶激活来改善2-DG诱发的细胞凋亡。方法通过细胞计数或MTT法测定细胞增殖。细胞周期,凋亡和坏死通过碘化丙啶染色和/或膜联蛋白V标记,然后通过流式细胞术确定。通过钙黄绿素-ATM或若丹明123标记后流式细胞术确定线粒体的孔过渡和去极化。通过二乙酸二氯二氢荧光素或一氯二苯胺标记,然后用流式细胞术或荧光法测定细胞内活性氧的种类和谷胱甘肽。通过Western印迹分析蛋白激酶的表达和磷酸化。结果(i)通过与亚致死浓度的类黄酮槲皮素共同处理人HL60急性粒细胞白血病细胞,大大增强了2-DG诱导的凋亡。考虑到数量上的差异,使用NB4早幼粒细胞和THP-1原单核细胞,使用姜黄素或染料木黄酮代替槲皮素,并使用lonidamine代替2-DG,但在通过葡萄糖-葡萄糖温育代替2-DG时,也获得了凋亡增强作用。免费媒体。 (ii)槲皮素和2-DG迅速引起线粒体孔转换的开放,这是在凋亡触发之前发生的。 (iii)处理不影响谷胱甘肽水平,对活性氧的产生产生不同的影响,与致死率的变化不匹配。 (iv)2-DG和lonidamine刺激防御性Akt和ERK磷酸化/激活,而葡萄糖饥饿则无效。多酚阻止了Akt磷酸化的刺激,在某些情况下还阻止了ERK的磷酸化。此外,槲皮素和2-DG刺激GSK-3α,β磷酸化/失活,尽管具有不同的同工型特异性。药理抑制剂的使用证实了这些激酶修饰对于细胞凋亡的重要性。结论目前的体外观察结果表明,低浓度选择的多酚类药物的联合治疗可能是改善某些糖酵解抑制剂抗肿瘤效果较差的一种方式,并且细胞凋亡的增强至少可以部分通过防御性调节来解释。蛋白激酶活性。

著录项

相似文献

  • 外文文献
  • 中文文献
  • 专利
获取原文

客服邮箱:kefu@zhangqiaokeyan.com

京公网安备:11010802029741号 ICP备案号:京ICP备15016152号-6 六维联合信息科技 (北京) 有限公司©版权所有
  • 客服微信

  • 服务号