Effects of 4-Hexylresorcinol on Protein Expressions in RAW 264.7 Cells as Determined by Immunoprecipitation High Performance Liquid Chromatography

Abstract

4-Hexylresorcinol (4HR) is a small organic compound that is used as an additive antiseptic and antioxidant, but its molecular properties have not been clearly elucidated. The present study explored the cellular effects of 4HR on RAW 264.7 cells by immunoprecipitation high-performance liquid chromatography (IP-HPLC) using 216 antisera. 4HR-treated cells showed significant decreases in the expressions of proliferation-related proteins, cMyc/MAX/MAD network, p53/Rb/E2F and Wnt/β-catenin signalings, epigenetic modifications, and protein translation. Furthermore, 4HR suppressed the expressions of growth factors and proteins associated with RAS signaling, NFkB signaling, inflammation, and osteogenesis, but elevated the expressions of proteins associated with p53-mediated and FAS-mediated apoptosis, T-cell immunity, angiogenesis, antioxidant, and oncogenic signaling. In a 4HR adherence assay, TNFα, PKC, osteopontin, and GADD45 were strongly adherent to 4HR-coated beads, whereas IL-6, c-caspase 3, CDK4, and c-caspase 9 were not. Many 4HR adherent proteins were expressed at lower levels in 4HR treated RAW 264.7 cells than in non-treated controls, whereas 4HR non-adherent proteins were expressed at higher levels. These observations suggest 4HR affects the expressions of proteins in an adhesion-dependent manner and that its effects on proteins are characteristic and global in RAW 264.7 cells.

4-Hexylresorcinol Exhibits Different Characteristics to Estrogen

“…According to NIH toxicology report, 4HR has shown anticancer effects in various types of cancers [15]. As 4HR increases the apoptotic stress mediated by mitochondrial proteins [18], cancer cells seem to be more influenced by 4HR administration. Based on these findings, there has been no evidence that 4HR might increase the incidence of breast cancer.…”

“…In this study, any clue of carcinogenic effect by 4HR on breast cancer cells was not found. In addition, we recently clarified the effect of angiogenesis and osteogenesis by 4HR is mediated not by estrogen, but by transforming growth factor-β1 [18,36].…”

“…However, there has been no convincing evidence for 4HR as a xenoestrogen. 4HR can induce cellular apoptosis in a dose-dependent and cellular type-dependent manner [4,18]. Thus, the inhibition of the spermatogenesis may be a complication of lethal dosage.…”

Immunomodulation for maxillofacial reconstructive surgery

“…5). 4HR administered to M2-type macrophages expresses a high level of TGF-β1 [63,64]. Osteal macrophages (OsteoMacs) produce osteoblast activators, such as TGF-β [65], osteopontin [66], and bone morphogenetic protein 2 (BMP-2) [67].…”

“…TGF-β1 is an important cytokine in wound healing [79]. Though the downstream signals from TGF-β1 are divergent, 4HR decreases TGF-β1 associated inflammatory reactions by inhibiting the NF-κB pathway [63]. In the diabetic animal model, 4HR ointment showed a substantial increase in capillary regeneration as compared to the untreated control (data submitted for publication).…”