Department of Pathology and Johns Hopkins Cancer Center, Johns Hopkins University School of Medicine, Baltimore, Maryland 21231, USA.
J Biol Chem. 2011 Sep 9;286(36):31457-65. doi: 10.1074/jbc.M111.274308. Epub 2011 Jul 16.
The efficacy of drugs used to treat cancer can be significantly attenuated by adaptive responses of neoplastic cells to drug-induced stress. To determine how cancer cells respond to inhibition of the enzyme fatty acid synthase (FAS), we focused on NF-κB-mediated pathways, which can be activated by various cellular stresses. Treating lung cancer cells with C93, a pharmacological inhibitor of FAS, results in changes indicative of a rapid initiation of NF-κB signaling, including translocation of RelA/p65 NF-κB to the nucleus, activation of a transfected NF-κB-luciferase reporter, and increased expression of NF-κB-dependent transcripts, IL-6, IL-8, and COX-2. Verifying that these responses to C93 are specifically related to inhibition of FAS, we confirmed that levels of these same transcripts increase in response to siRNA targeting FAS. Inhibiting this NF-κB response (either by transfecting a mutant IκBα or treating with bortezomib) resulted in increased cell killing by C93, indicating that the NF-κB response is protective in this setting. Because inhibiting FAS leads to accumulation of intermediate metabolites of fatty acid biosynthesis, we then questioned whether protein kinase C (PKC) is involved in this response to metabolic stress. Immunofluorescence microscopy revealed that C93 treatment results in cellular translocation of PKCα and PKCβ isoforms and increased PKCα-dependent phosphorylation of the IκBα subunit of NF-κB. Furthermore, inhibiting PKC activity with RO-31-8220 or PKCα isoform-specific siRNA attenuates C93-induced IκBα phosphorylation and NF-κB activation and also potentiates C93-induced cell killing. These results suggest a link between PKC and NF-κB in protecting cancer cells from metabolic stress induced by inhibiting FAS.
用于治疗癌症的药物的疗效可能会因肿瘤细胞对药物诱导的应激的适应性反应而显著降低。为了确定癌细胞如何对脂肪酸合酶(FAS)的抑制剂的抑制作用作出反应,我们集中研究了 NF-κB 介导的途径,该途径可被各种细胞应激激活。用 C93(一种 FAS 的药理学抑制剂)处理肺癌细胞会导致 NF-κB 信号快速起始的变化,包括 RelA/p65 NF-κB 向核内易位、转染的 NF-κB-荧光素酶报告基因的激活以及 NF-κB 依赖性转录物、IL-6 和 IL-8 和 COX-2 的表达增加。验证这些对 C93 的反应与 FAS 的抑制特异性相关,我们证实了相同的转录物水平会因针对 FAS 的 siRNA 而增加。抑制这种 NF-κB 反应(通过转染突变型 IκBα或用硼替佐米治疗)会导致 C93 的细胞杀伤增加,表明在这种情况下 NF-κB 反应具有保护作用。因为抑制 FAS 会导致脂肪酸生物合成的中间代谢物的积累,所以我们随后询问蛋白激酶 C(PKC)是否参与这种代谢应激反应。免疫荧光显微镜显示,C93 处理导致 PKCα 和 PKCβ 同工型的细胞内易位和 NF-κB 的 IκBα亚单位的 PKCα依赖性磷酸化增加。此外,用 RO-31-8220 或 PKCα 同工型特异性 siRNA 抑制 PKC 活性会减弱 C93 诱导的 IκBα磷酸化和 NF-κB 激活,并增强 C93 诱导的细胞杀伤。这些结果表明 PKC 和 NF-κB 之间存在联系,可保护癌细胞免受抑制 FAS 诱导的代谢应激的影响。