β-Hydroxy-β-methylbutyrate Suppresses NF-ĸB Activation and IL-6 Production in TE-1 Cancer Cells

Abstract

Background/Aim: Stress reactions, especially those related to surgery, cause poor convalescence of cancer patients. β-Hydroxyβ-methylbutyrate (HMB) is known to regulate excessive inflammation in the body. The objective of this work was to investigate the capacity of HMB to suppress activation of nuclear factor-kappa B (NF-ĸB) and production of interleukin-6 (IL-6) in a human esophageal squamous cell carcinoma cell line . Materials and Methods: Cell proliferation was measured using the water-soluble tetrazolium-1 method, while tumor necrosis factor alpha (TNFα)-induced IL-6 production was measured using an enzyme-linked immunosorbent assay (ELISA) assay. Nuclear translocation of NF-ĸB was detected by immunofluorescence staining. Results: HMB did not affect cell proliferation. However, HMB suppressed the TNFα-induced increase in IL-6 production in TE-1 cells by inhibiting NF-ĸB activation. Conclusion: HMB did not influence TE-1 cell proliferation, but inhibited activation of NF-ĸB and IL-6 production. This result may be useful for improving excessive stress reactions during and after surgery.During and after cancer surgery, operative stress can promote tumor metastasis and result in poor prognoses. Notably, stress related to thoraco-laparotomy for esophageal cancer carries an extremely high risk of postoperative complications. Excessive post-surgical stress or postoperative complications, so-called second attacks, cause a cytokine storm that promotes tumor metastasis (surgical oncotaxis). Reported mechanisms underlying surgical oncotaxis include increased adrenal corticoid levels, blood coagulability, immunosuppression, and reactive oxygen species production (1-3).Usually, increased levels of cytokines, such as interleukin-1 (IL-1) or tumor necrosis factor alpha (TNFα), are observed several hours after surgery, followed by increased IL-6 or IL-8 (4, 5). TNFα, a representative inflammatory cytokine secreted in the tumor microenvironment, elicits secretion of other cytokines by activating nuclear factor-kappa B (NF-ĸB) signaling (6). NF-ĸB plays a central role in inflammatory reactions, and is the primary activator of cytokine storms (7,8). Target genes of NF-ĸB include adhesion factors, iNOS (inducible nitric oxide synthase), and other factors (9). Thus, NF-ĸB plays an important role in tumor survival, metastasis, and chemotherapy resistance. NF-ĸB is essential for promoting inflammation-related cancer. This mechanism can significantly impact chemotherapy and cancer-related surgeries (10). Therefore, NF-ĸB regulation is extremely important for cancer therapies.β-hydroxyβ-methylbutyrate (HMB), a metabolite of the branched-chain amino acid leucine that obstructs NF-ĸB (p65) function, has attracted attention for its ability to control excessive inflammatory reactions. HMB reportedly reduces inflammatory cytokine production, apoptosis, and activity of proteolysis-inducing factor (11). Thus, we hypothesized that HMB regulates activation of the ubiquitin-proteasome pathway to control NF-kB activation and excessive inflamm...

The Effects of β-Hydroxy-β-Methylbutyrate (HMB) on Chemotaxis, Phagocytosis, and Oxidative Burst of Peripheral Blood Granulocytes and Monocytes in Goats

“…According to Townsend et al [45], in men, dietary supplementation with HMB can decrease the expression of tumour necrosis factor alpha (TNF-α) and TNF-α receptor 1 (TNFR1) on CD14+ monocytes measured by flow cytometry in circulating blood. In a study by Miyake et al [46], HMB did not influence TE-1 cell proliferation, but it inhibited the production of NF-kB and IL-6 on the TE-1 cancer cell line in vitro. Hoffman et al [47] demonstrated that 23 days of HMB supplementation in combat soldiers can attenuate the inflammatory response (TNF-α, interleukins IL-8 and IL-10, granulocyte colony-stimulating factor (GC-SF), interferon (IFN)-γ, and fractalkine) to intense military training, and maintain muscle quality.…”

The effects of preoperative supplementation with a combination of beta‐hydroxy‐beta‐methylbutyrate, arginine, and glutamine on inflammatory and hematological markers of patients with heart surgery: a randomized controlled trial

“…HMB's biological effects include changes in protein production, inhibition of proteolysis, and anti-inflammatory actions, which help to preserve muscle mass in both young and old subjects [ 40 , 41 ]. Miyake and coworkers were evaluated the anti-inflammatory effects of HMB in TE-1 cancer cells and reported that HMB administration could downregulate IL-6 production by regulation of inhibitor of kappa B alpha (IĸBα) phosphorylation and reduction of nuclear translocation of NF-ĸB/Rela [ 42 ].…”

The influence of dietary supplementation with the leucine metabolite β-hydroxy-β-methylbutyrate (HMB) on chemotaxis, phagocytosis and respiratory burst of peripheral blood granulocytes and monocytes in calves

“…In a study by Townsend et al [53], a flow cytometry analysis revealed that the circulating levels of TNF-α (tumour necrosis factor) and TNF-α receptor 1 (TNFR1) in CD14 + monocytes decreased in men whose diets were supplemented with HMB. In an in vitro study of the TE-1 cancer cell line, Miyake et al [54] demonstrated that HMB inhibited the activation of NF-ĸB and IL-6 production. In a study by Hoffman et al [55], the inflammatory response (TNF-α, interleukin (IL)-8, IL-10, granulocyte colony-stimulating factor (GC-SF), interferon (IFN)-γ, and fractalkine) to intense military training was attenuated and muscle quality was maintained in combat soldiers whose diets were supplemented with HMB for 23 days.…”