Department of Pathology, Northwestern University, Chicago, IL 60613, USA.
Mol Cell Biochem. 2011 Jun;352(1-2):209-19. doi: 10.1007/s11010-011-0756-y. Epub 2011 Feb 26.
Microcystin-LR (MC-LR) is the most frequent and most toxic microcystin identified. This natural toxin has multiple features, including inhibitor of protein phosphatases 1 and 2A, inducer of oxidative stress, as well as, tumor initiator and promoter. One unique character of MC-LR is this chemical can accumulate into liver after contacting and lead to severe damage to hepatocytes, such as apoptosis. Fas receptor (Fas) and Fas ligand (FasL) system is a critical signaling system initiating apoptosis. In current study, we explored whether MC-LR could induce Fas and FasL expression in HepG2 cells, a well used in vitro model for the study of human hepatocytes. The data showed MC-LR induced Fas and FasL expression, at both mRNA and protein levels. We also found MC-LR induced apoptosis at the same incubation condition at which it induced Fas and FasL expression. The data also revealed MC-LR promoted nuclear translocation and activation of p65 subunit of NF-κB. By applying siRNA to knock down p65 in HepG2 cells, we successfully impaired the activation of NF-κB by MC-LR. In these p65 knockdown cells, we also observed significant reduction of MC-LR-induced Fas expression, FasL expression, and apoptosis. These findings demonstrate that the NF-κB mediates the induction of Fas and FasL as well as cellular apoptosis by MC-LR in HepG2 cells. The results bring important information for understanding how MC-LR induces apoptosis in hepatocytes.
微囊藻毒素-LR(MC-LR)是最常见和最具毒性的微囊藻毒素。这种天然毒素具有多种特性,包括蛋白磷酸酶 1 和 2A 的抑制剂、氧化应激诱导剂,以及肿瘤起始剂和促进剂。MC-LR 的一个独特特征是,这种化学物质在接触后会积聚到肝脏中,导致肝细胞严重损伤,如细胞凋亡。Fas 受体(Fas)和 Fas 配体(FasL)系统是启动细胞凋亡的关键信号系统。在目前的研究中,我们探讨了 MC-LR 是否可以诱导 HepG2 细胞中 Fas 和 FasL 的表达,HepG2 细胞是一种常用于研究人类肝细胞的体外模型。研究数据表明,MC-LR 可诱导 Fas 和 FasL 在 mRNA 和蛋白质水平上的表达。我们还发现,在诱导 Fas 和 FasL 表达的相同孵育条件下,MC-LR 也可诱导细胞凋亡。研究数据还揭示,MC-LR 可促进 NF-κB 的 p65 亚基向核内易位和激活。通过在 HepG2 细胞中应用 siRNA 敲低 p65,我们成功地削弱了 MC-LR 对 NF-κB 的激活。在这些敲低 p65 的细胞中,我们还观察到 MC-LR 诱导的 Fas 表达、FasL 表达和细胞凋亡显著减少。这些发现表明,NF-κB 介导了 MC-LR 在 HepG2 细胞中诱导 Fas 和 FasL 以及细胞凋亡。这些结果为了解 MC-LR 如何诱导肝细胞凋亡提供了重要信息。