The IKK complex, or inhibitor of NFkB kinase, is composed of IKK alpha and IKK beta. These kinases are at the core of the NFkB signaling cascade. The NFkB family is made up of transcription factors that are kept inactive in the cytoplasm through inhibitory IkB proteins. When various stimuli are activated, these IKK proteins go through phosphorylation, leading to polyubiquitination and subsequent degradation. Without the presence of these inhibitory factors, NFkB can now enter the nucleus and turn on the transcription of a variety of genes that influence cellular proliferation, immune response, apoptosis, cancer pathways and more. Recently, research has shed light on the yellow pigment from Curcuma longa, curcumin, as having potential therapeutic properties through its antioxidant and anti-inflammatory characteristics. The following experiments use an IKB alpha antibody to investigate the effects of curcumin on the NFkB signaling pathway.
Simple Western: IkB-alpha Antibody (6A920) [NB100-56507] - Simple Western lane view shows a specific band for IKB alpha in 0.5 mg/ml of NIH-3T3 lysate. This experiment was performed under reducing conditions using the 12-230 kDa separation system.
Kasinski et al used an IKB alpha antibody to study the effects that a curcumin monoketone analog (EF24) has on the inhibition of the NFkB signaling pathway. Initially testing on this curcumin compound exhibited potent anticancer behavior. First, it was established that the curcumin analog had more potent cell toxicity than curcumin itself in ovarian, breast, prostate and cervical cancer cells. Once this was determined, immunoprecipitation studies using an IKB alpha antibody showed that EF24 inhibits TNF alpha induced IKB phosphorylation. What’s more, A549 cells pretreated with EF24 or curcumin and then stimulated by TNF-alpha had interesting results. An IKB alpha antibody in western blot showed that TNF alpha alone induced IKB alpha degradation, where the treatment of EF24 blocked this effect.
Furthermore, Bimonte et al used an IKB alpha antibody to take a closer look at the curcumin compound and its effect on tumor growth and angiogenesis in pancreatic cancer. The curcumin compound was of initial interest because it has similar protective effects as the pancreatic cancer chemotherapeutic agent gemcitabine without the side effects. First, an IKB alpha antibody was used in western blot on a mouse model of pancreatic cancer that was generated via injection of MIA PaCa-2-RFP cells into the pancreas of nude mice. The western blot results indicated that there was a reduced expression of IKB alpha and IKB beta in the tumor of mice treated with curcumin versus that of controls. Overall, the previous two studies demonstrate that curcumin and its analog EF24 are successful at changing the expression levels of IKB alpha and beta through the NFkB pathway.
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