■已经显示橙皮苷通过抑制核因子-κB(NF-κB)活化来诱导NALM-6细胞的凋亡。
■为了研究橙皮苷通过阻断磷酸肌醇3-激酶(PI3K)/Akt通路作为癌症治疗的主要靶点对NF-κB活化的抑制作用,在NALM-6细胞中。
■NALM-6细胞与两种浓度的橙皮苷(25,50μM)在存在或不存在胰岛素(100nM)的情况下孵育,作为Akt的有效激活剂。橙皮苷的细胞毒性活性通过3-(4,5-甲基噻唑-2-基)-2,5-二苯基四唑溴化物测定法测定。使用细胞死亡检测ELISAPlus试剂盒通过ELISA测试测量细胞凋亡死亡。为了检测橙皮苷对Akt通路的影响,Akt的磷酸化水平,κBα(IκBα)抑制剂,通过Westernblot分析确定糖原合酶激酶-3β(GSK-3β)和IκB激酶α(IKKα)的表达水平。
■橙皮苷(两种浓度)与未处理的细胞相比,在存在和不存在胰岛素的情况下以时间依赖性方式显著降低细胞存活(P<0.05)。即使在高胰岛素血症条件下,橙皮苷也显着增加NALM-6细胞的凋亡(P<0.0001)。橙皮苷抑制胰岛素诱导的磷酸化和Akt的激活,IκBα,GSK-3β和IKKα表达降低。
■这项研究的结果表明,橙皮苷的细胞毒性和促凋亡作用部分是通过抑制PI3K3/Akt/IKK信号通路介导的。所以,橙皮苷可能通过靶向细胞存活途径作为化疗药物。
UNASSIGNED: It has been shown that hesperidin induces apoptosis in NALM-6 cells through inhibition of nuclear factor-kappa B (NF-κB) activation.
UNASSIGNED: To investigate the effect of hesperidin on inhibition of NF-κB activation through blocking phosphoinositide 3-kinase (PI3K)/Akt pathway as a main target in cancer treatment, in NALM-6 cells.
UNASSIGNED: NALM-6 cells were incubated with two concentrations of hesperidin (25, 50 μM) in the presence or absence of insulin (100 nM), as a potent activator of Akt. The cytotoxic activity of hesperidin was determined by 3-(4,5-methylthiazol-2-yl)-2,5-diphenyltetrazolium bromide assay. Cell apoptotic death was measured by ELISA test using cell death detection ELISA Plus kit. To assay the effect of hesperidin on Akt pathway, the phosphorylation levels of Akt, inhibitor of kappa B alpha (IκBα), and glycogen synthase kinase-3 beta (GSK-3β) and expression level of IκB kinase alpha (IKKα) were determined by Western blot analysis.
UNASSIGNED: Hesperidin (both concentrations) significantly reduced cells survival in the presence and absence of insulin compared to untreated cells in a time-dependent manner (P < 0.05). Hesperidin also significantly increased apoptosis in NALM-6 cells even in hyperinsulinemia condition (P < 0.0001). Hesperidin inhibited insulin-induced phosphorylation and activation of Akt, IκBα, and GSK-3β and decreased expression of IKKα.
UNASSIGNED: The results of this study demonstrated that cytotoxic and proapoptotic actions of hesperidin are partly mediated through the suppression of PI3K3/Akt/IKK signaling pathway. So, hesperidin might act as a chemotherapeutic agent by targeting cell survival pathways.