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Induction of CYP4F11 by cytokines through the JNK pathway. a, effects of cytokines and TPA on CYP4F11 transcripts in HaCaT cells. Cells were treated with 10 ng/ml TNF-, 10 ng/ml IL-1, or 100 ng/ml TPA for 24 h. Cells treated with 0.1% DMSO or 0.1% bovine serum albumin were used as the vehicle control. The expression of CYP4F11 was quantitated by qRT-PCR. Each data point represents n 5. , P 0.05, , P 0.01 compared with the control group. b and c, effects of pathway inhibitors on the induction of CYP4F11 by cytokines. HaCaT cells preincubated with or without SP600125 (JNK inhibitor) or SB203580 (p38 inhibitor) were treated with 10 ng/ml TNF-(b) or 10 ng/ml IL-1 (c). At 24 h post-treatment, total RNA was isolated, and the expression of CYP4F11 was quantitated by qRT-PCR. Cells preincubated with 0.1% DMSO and then treated with 0.1% bovine serum albumin were used as the vehicle control. Each data point represents n 5. , P 0.01 compared with the relative control group. d, TPA had no effect on c-Jun phosphorylation in HaCaT cells. Cells were treated with 100 ng/ml TPA and collected at different time points as indicated in the figure. Whole cell lysate was isolated from each sample, and 20 g of proteins from each sample was loaded for Western blot assay. -Actin was used as the loading control. e, effects of TNF-on c-Jun phosphorylation in HaCaT cells. Cells preincubated with or without 20 M SP600125 were treated with 10 ng/ml TNF-and collected at different time points as indicated in the figure. Whole cell lysate was isolated from each sample, and 15 g of proteins from each sample was loaded for Western blot assay. -Actin was used as the loading control. f, inhibition effects of SP600125, 9-cis-retinoic acid, and all-trans-retinoic acid on TNF--induced c-Jun phosphorylation in HaCaT cells. Cells preincubated with or without 20 M SP600125, 1 M 9-cis-retinoic acid, or 1 M all-trans-retinoic acid were treated with 10 ng/ml TNF-and collected at different time points as indicated in the figure. Whole cell lysate was isolated from each sample, and 15 g of proteins from each sample was loaded for Western blot assay. -Actin was used as the loading control.
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Mechanisms regulating CYP4F genes remain under investigation, although characterization of CYP4F regulatory modalities would facilitate the discovery of new drug targets. This present study shows that all-trans- and 9-cis-retinoic acids can inhibit CYP4F11 expression in human keratinocyte-derived HaCaT cells. Transrepression of many genes by retino...
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Retinoid X receptors (RXRs) are heterodimerization partners for many nuclear receptors and also act as homodimers. Heterodimers formed by RXR and a nonpermissive partner, e.g. retinoic acid receptor (RAR) and vitamin D receptor (VDR), can be activated only by the agonist of the partner receptor. In contrast, heterodimers that contain permissive par...
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... For example, it is known that CYP4 enzymes allow the inactivation of HETEs by ωhydroxylation [16,17], but their involvement in immune processes is mostly studied in arachidonic acid ω-hydroxylation to 20-HETE or leukotriene metabolism [18][19][20]. The involvement of the nuclear receptor RXR and NF-kB in CYP4F11 expression has been described in immortalized keratinocyte and hepatocyte lines [21,22]. In another study, some CYP4 family enzymes have been studied in polymorphonuclear leukocytes (neutrophils), but, specifically, CYP4F11 was not considered by authors [23]. ...
Malaria is a frequent parasitic infection becomes life threatening due to the disequilibrated immune responses of the host. Avid phagocytosis of malarial pigment hemozoin (HZ) and HZ-containing Plasmodium parasites incapacitates monocyte functions by bioactive lipoperoxidation products 4-hydroxynonenal (4-HNE) and hydroxyeicosatetraenoic acids (HETEs). CYP4F conjugation with 4-HNE is hypothesised to inhibit ω-hydroxylation of 15-HETE, leading to sustained monocyte dysfunction caused by 15-HETE accumulation. A combined immunochemical and mass-spectrometric approach identified 4-HNE-conjugated CYP4F11 in primary human HZ-laden and 4-HNE-treated monocytes. Six distinct 4-HNE-modified amino acid residues were revealed, of which C260 and H261 are localized in the substrate recognition site of CYP4F11. Functional consequences of enzyme modification were investigated on purified human CYP4F11. Palmitic acid, arachidonic acid, 12-HETE, and 15-HETE bound to unconjugated CYP4F11 with apparent dissociation constants of 52, 98, 38, and 73 µM, respectively, while in vitro conjugation with 4-HNE completely blocked substrate binding and enzymatic activity of CYP4F11. Gas chromatographic product profiles confirmed that unmodified CYP4F11 catalysed the ω-hydroxylation while 4-HNE-conjugated CYP4F11 did not. The 15-HETE dose dependently recapitulated the inhibition of the oxidative burst and dendritic cell differentiation by HZ. The inhibition of CYP4F11 by 4-HNE with consequent accumulation of 15-HETE is supposed to be a crucial step in immune suppression in monocytes and immune imbalance in malaria.
... Wang et al. [117] demonstrated that CYP4F11 expression is mediated by RXR and c-Jun N-terminal kinase (JNK) signaling pathways in human keratinocyte-derived cells (HaCaT). While treatment with tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) increases CYP4AF11 levels by the interaction between the activating protein-1 (AP-1) complex and JNK pathway, and retinoids have an inhibitory effect in CYP4F11 induction [117]. ...
... Wang et al. [117] demonstrated that CYP4F11 expression is mediated by RXR and c-Jun N-terminal kinase (JNK) signaling pathways in human keratinocyte-derived cells (HaCaT). While treatment with tumor necrosis factor α (TNF-α) and interleukin 1β (IL-1β) increases CYP4AF11 levels by the interaction between the activating protein-1 (AP-1) complex and JNK pathway, and retinoids have an inhibitory effect in CYP4F11 induction [117]. A similar inhibitory effect in CYP4F11 expression is also observed in HepG2 cells through the modulation of the nuclear factor-kappa B (NF-kB) pathway by TNF-α and mitogen-activated protein kinase kinase kinase (MEKK) overexpression [118]. ...
Orphan cytochromes P450 (CYP) are enzymes whose biological functions and substrates are unknown. However, the use of new experimental strategies has allowed obtaining more information about their relevance in the metabolism of endogenous and exogenous compounds. Likewise, the modulation of their expression and activity has been associated with pathogenesis and prognosis in different diseases. In this work, we review the regulatory pathways and the possible role of orphan CYP to provide evidence that allow us to stop considering some of them as orphan enzymes and to propose them as possible therapeutic targets in the design of new strategies for the treatment of diseases associated with CYP-mediated metabolism.
... Among the all differentially expressed genes, IGFN1, CYP4F11, KCNN3, CYP26A1, and ATP8A1 were much more significant up or downregulated genes, which mainly participated in cell metabolism processes. For instance, CYP4F11, and CYP26A1 primarily catalyze many reactions involved in drug metabolism, synthesis of cholesterol, steroids and other lipids (Wang et al., 2010;Sun et al., 2015;Qiu et al., 2017). KCNN3 and ATP8A1 are mainly responsible for ions metabolism, including the activation and transport of potassium ion, calcium ion and heavy metal ion (Jing et al., 2019;Rahm et al., 2021). ...
With the rapid development of nanotechnology and nanoscience, nanosafety assessment has raised public concern. Although many studies have illustrated that nanomaterials could lead to genotoxicity, the early alterations of DNA methylation with nanomaterials under low-dose exposure have not been completely clear. In this study, we investigated the potential effect and molecular mechanism of AgNPs on the alternation of DNA methylation fingerprints in HEK293T cells under sublethal exposure. Intriguingly, silver nanoparticle treatment increased 5-mC level and changed methylation-related enzyme contents. Mechanistically, we scrutinized the changes in the molecular signaling and biological functions by means of MeDIP-Seq and RNA-seq. Our results revealed that AgNPs might undermine a number of vital regulatory networks including the metabolic processes, biological regulation and other cellular processes. More specifically at the DNA methylation fingerprints, there were 12 up-regulated and simultaneous hypomethylated genes, and 22 down-regulated and concomitant hypermethylated genes in HEK293T cells responding to AgNPs. Notably, these genes were primarily involved in lipid metabolism and ion metabolism. Together, these responsive genes might be used as early sensitive indicators for the variations of early epigenetic integrity through changing the DNA methylation fingerprints, as reflective of biological risk and toxicity of silver nanoparticles under realistic exposure scenarios.
... Cells were treated with H 2 O 2 (200µM) in the presence and absence of inhibitors, i.e. SB203580 (20mM) for p38MAPK (37), SP600125 (20mM) for JNK (56), Ro 31-7549 (2mM) and Go6976 (1mM) for PKC (57). Again, they were treated with inhibitors 1 hour before addition of H 2 O 2 and incubated for additional 4 hours before fixation. ...
Pemphigus Vulgaris (PV) is a life-threatening autoimmune disease manifested with blisters in the skin and mucosa and caused by autoantibodies against adhesion protein desmoglein-3 (Dsg3) expressed in epithelial membrane linings of these tissues. Despite many studies, the pathogenesis of PV remains incompletely understood. Recently we have shown Dsg3 plays a role in regulating the yes-associated protein (YAP), a co-transcription factor and mechanical sensor, and constraining reactive oxygen species (ROS). This study investigated the effect of PV sera as well as the anti-Dsg3 antibody AK23 on these molecules. We detected elevated YAP steady-state protein levels in PV cells surrounding blisters and perilesional regions and in keratinocytes treated with PV sera and AK23 with concomitant transient ROS overproduction. Cells treated with hydrogen peroxide also exhibited augmented nuclear YAP accompanied by reduction of Dsg3 and α-catenin, a negative regulator of YAP. As expected, transfection of α-catenin-GFP plasmid rendered YAP export from the nucleus evoked by hydrogen peroxide. In addition, suppression of total YAP was observed in hydrogen peroxide treated cells exposed to antioxidants with enhanced cell-cell adhesion being confirmed by decreased fragmentation in the dispase assay compared to hydrogen peroxide treatment alone. On the other hand, the expression of exogenous YAP disrupted intercellular junction assembly. In contrast, YAP depletion resulted in an inverse effect with augmented expression of junction assembly proteins, including Dsg3 and α-catenin capable of abolishing the effect of AK23 on Dsg3 expression. Finally, inhibition of other kinase pathways, including p38MAPK, also demonstrated suppression of YAP induced by hydrogen peroxide. Furthermore, antioxidant treatment of keratinocytes suppressed PV sera-induced total YAP accumulation. In conclusion, this study suggests that oxidative stress coupled with YAP dysregulation attributes to PV blistering, implying antioxidants may be beneficial in the treatment of PV.
... Articles differential genes in response to treatment with ATRA and 9cis-retinoic acid in these cells. 46 The fluorescence emission from the compound could be detected at around 450−480 nm when DC271 (4c) was excited with 365−405 nm light ( Figure 7). The distribution of the fluorescence signal varied somewhat depending on compound concentration and the duration of treatment; however, in general, the compound could be detected diffusely in the nucleus and in some brightly fluorescent, punctate structures in the cytoplasm (presumably lipid vesicles). ...
Retinoids, such as all-trans-retinoic acid (ATRA), are endogenous signalling molecules derived from Vitamin A that influ-ence a variety of cellular processes through mediation of transcription events in the cell nucleus. Due to these wide-ranging and powerful biological activities, retinoids have emerged as therapeutic candidates of enormous potential. However, their use has been limited, to date, due to a lack of understanding of the complex and intricate signaling pathways that they con-trol. We have designed and synthesized a family of synthetic retinoids that exhibit strong, intrinsic, solvatochromatic fluo-rescence as multifunctional tools to interrogate these important biological activities. We utilized the unique photophysical characteristics of these fluorescent retinoids to develop a novel in vitro fluorometric binding assay to characterize and quanti-fy their binding to their cellular targets, including Cellular Retinoid Binding Protein II (CRABPII). The dihydroquinoline retinoid, DC360, exhibited particularly strong binding (Kd = 34.0 ± 2.5 nM) and we further used X-ray crystallography to solve the structure of the DC360-CRABPII complex to 1.8 Å, which showed that DC360 occupies the known hydrophobic retinoid-binding pocket. Finally, we used confocal fluorescence microscopy to image the cellular behaviour of the com-pounds in cultured human epithelial cells, highlighting a fascinating nuclear localisation, and used RNA sequencing to con-firm that the compounds regulate similar cellular processes to ATRA. We anticipate that the unique properties of these fluo-rescent retinoids can now be used to shed new light on the vital and highly complex retinoid signalling pathway.
... Previous studies showed that increase of TNF-α-induced CYP4F11 was in response to jun N-terminal protein kinase (JNK) pathway activity (Sechler et al., 2013). Another finding suggested that the CYP ω-hydroxylase, consisting of CYP4A and CYP4F molecules, promoted angiogenesis and metastatic potential in human NSCLC cells through up-regulation of Vascular endothelial growth factor (VEGF) and Matrix metallopeptidase 9 (MMP-9) (Wang et al., 2010;Bell and Strobel, 2012). Previously, CYP2D6 was found to be another important CYP450 enzyme involved in drug responses, accounting for 30% of metabolism of all medications (Gurley et al., 2008). ...
The efficacy of erlotinib treatment for advanced non-small cell lung cancer (NSCLC) is due to its action as an epidermal growth factor receptor tyrosine kinase inhibitor (EGFR-TKI). Patients treated with erlotinib experience different drug responses. The effect of germline mutations on therapeutic responses and adverse drug responses (ADRs) to erlotinib in Chinese patients requires elucidation. Sixty Han Chinese advanced non-small cell lung cancer patients received erlotinib monotherapy and, for each participant, 76 candidate genes (related to EGFR signaling, drug metabolism and drug transport pathways) were sequenced and analyzed. The single-nucleotide polymorphisms (SNPs) rs1042640 in UGT1A10, rs1060463, and rs1064796 in CYP4F11, and rs2074900 in CYP4F2 were significantly associated with therapeutic responses to erlotinib. Rs1064796 in CYP4F11 and rs10045685 in UGT3A1 were significantly associated with adverse drug reaction. Moreover, analysis of a validation cohort confirmed the significant association between rs10045685 in UGT3A1 and erlotinib adverse drug response(unadjusted p = 0.015). This study provides comprehensive, systematic analyses of genetic variants associated with responses to erlotinib in Chinese advanced non-small cell lung cancer patients. Newly-identified SNPs may serve as promising markers to predict responses and safety in erlotinib-treated advanced non-small cell lung cancer patients after chemotherapy doublet.
... Underexpression of EPHX1 is frequently described in COPD airway and lung tissues, and "slow" EPHX1 SNPs are associated with impaired enzyme activity and increased COPD risk, and "fast" SNPs potentially confer a protective effect (40). Cytochrome P450 4F enzymes, such as CYP4F11, are involved in cellular protection, xenobiotic metabolism, detoxification, lipid synthesis, and metabolic activation of drugs, including those used to treat chronic inflammatory disease (63,64). They also have a direct role in inhibiting inflammation through suppression of leukotriene and prostaglandin signals (64). ...
... Cytochrome P450 4F enzymes, such as CYP4F11, are involved in cellular protection, xenobiotic metabolism, detoxification, lipid synthesis, and metabolic activation of drugs, including those used to treat chronic inflammatory disease (63,64). They also have a direct role in inhibiting inflammation through suppression of leukotriene and prostaglandin signals (64). CYP4F11 contains both c-Jun N-terminal kinase (JNK)/activator protein 1 and hormone response element binding domains; it is positively regulated by retinoid X receptors and JNK (through TNF-a activation), and is negatively regulated by retinoic acid receptors. ...
... However, regulation of CYP4F11 in an environment of chronic inflammation is complex. In human epidermal keratinocytes, although TNF-a leads to immediate activation of CYP4F11 through JNK, subsequent activation of NF-kb results in direct inhibition of CYP4F11 (63,64). CYP4F11 regulators are clearly important to COPD biology, although little is known about the function of this enzyme in respiratory tissues. ...
Rationale: DNA methylation is an epigenetic modification that is highly disrupted in response to cigarette smoke and involved in a wide spectrum of malignant and non-malignant diseases, but surprisingly not previously assessed in small airways of patients with chronic obstructive pulmonary disease (COPD). Small airways are the primary sites of airflow obstruction in COPD. We sought to determine whether DNA methylation patterns are disrupted in small airway epithelia of COPD patients, and evaluate whether changes in gene expression are associated with these disruptions. Methods: Genome-wide methylation and gene expression analysis were performed on small airway epithelial DNA and RNA obtained from the same patient during bronchoscopy, using Illumina's Infinium HM27 and Affymetrix's Genechip Human Gene 1.0 ST arrays. To control for known effects of cigarette smoking on DNA methylation, methylation and gene expression profiles were compared between former smokers (FS) with and without COPD matched for age, pack years and years of smoking cessation. Results: Our results indicate that aberrant DNA methylation is i) a genome-wide phenomenon in small airways of patients with COPD and ii) associated with altered expression of genes and pathways important to COPD, such as the Nrf2 oxidative response pathway. Conclusions: DNA methylation is likely an important mechanism contributing to modulation of genes important to COPD pathology. Since these methylation events may underlie disease-specific gene-expression changes, their characterization is a critical first step towards the development of epigenetic markers and an opportunity for developing novel epigenetic therapeutic interventions for COPD.
... They inactivate the leukotriene and prostaglandin prompts for the inflammation cascade playing an anti-inflammatory role, and they also catalyze the metabolism of many drugs (Hashizume et al., 2002;Kalsotra et al., 2004). Among the human CYP4F enzymes, CYP4F11 is most active in metabolizing therapeutic drugs and has been demonstrated that retinoids downregulate CYP4F11 expression in HaCaT cells for its anti AP-1 activity and supports the positive regulation of this cytochrome through the AP-1 complex by inflammatory cytokines TNF-α and IL-1β in accord with the literature (Wang et al., 2010). CYP4F11 catalyzes N-hydroxylations of leukotriene-B4, arachidonic acid, lipoxin-A4, and 8hydroxyeicosatetraenoic acid (Wang et al., 2010). ...
... Among the human CYP4F enzymes, CYP4F11 is most active in metabolizing therapeutic drugs and has been demonstrated that retinoids downregulate CYP4F11 expression in HaCaT cells for its anti AP-1 activity and supports the positive regulation of this cytochrome through the AP-1 complex by inflammatory cytokines TNF-α and IL-1β in accord with the literature (Wang et al., 2010). CYP4F11 catalyzes N-hydroxylations of leukotriene-B4, arachidonic acid, lipoxin-A4, and 8hydroxyeicosatetraenoic acid (Wang et al., 2010). ...
Involvement of cytochrome P450 genes (CYPs) in breast cancer (BCa) may differ between populations, with expression patterns affected by tumorigenesis. This may have an important role in the metabolism of anticancer drugs and in the progression of cancer. The aim of this study was to determine the mRNA expression patterns of four cytochrome P450 genes (CYP2W1, 3A5, 4F11 and 8A1) in Mexican women with breast cancer. Real- time PCR analyses were conducted on 32 sets of human breast tumors and adjacent non-tumor tissues, as well as 20 normal breast tissues. Expression levels were tested for association with clinical and pathological data of patients. We found higher gene expression of CYP2W1, CYP3A5, CYP4F11 in BCa than in adjacent tissues and only low in normal mammary glands in our Mexican population while CYP8A1 was only expressed in BCa and adjacent tissues. We found that Ki67 protein expression was associated with clinicopathological features as well as with CYP2W1, CYP4F11 and CYP8A1 but not with CYP3A5. The results indicated that breast cancer tissues may be better able to metabolize carcinogens and other xenobiotics to active species than normal or adjacent non-tumor tissues.
... TNF-stimulation causes an activation of two signal transduction pathways, the c-Jun NH 2 -terminal kinase (JNK) pathway and the nuclear factor B of the light-chain-enhancer in activated B cells (NF-B) pathway (De Smaele et al., 2001; Tang et al., 2001; Deng et al., 2003). Our previous studies have examined the roles the JNK pathway and retinoic acids play in the regulation of CYP4F11 and have found that JNK stimulation causes an increase in CYP4F11 mRNA, whereas retinoids cause down-regulation of CYP4F11 mRNA (Wang et al., 2010). However, we did not examine the effects of NF-B on CYP4F11 expression during TNF-stimulation. ...
... (Wang et al., 2010 ). TNF-activates two pathways: the JNK pathway , which was shown to up-regulate CYP4F11 expression through activator protein 1 (AP-1) binding sites, and the NF-B signaling pathway. ...
... However, what makes this finding unique is that the inhibition of endogenous CYP4F11 is not seen in the presence of TNF-after 24 h. This regulation by TNF-was reported in a previous study from our laboratory (Wang et al., 2010) and is verified in this study inFig. 1, which also shows that the presence of TNF-and an inhibitor of NF-B cause a greater increase in CYP4F11 transcript quantity than TNF-alone. ...
Although the mechanisms that regulate CYP4F genes have been and are currently being studied in a number of laboratories, the specific mechanisms for the regulation of these genes are not yet fully understood. This study shows that nuclear factor κB of the light-chain-enhancer in activated B cells (NF-κB) can inhibit CYP4F11 expression in human liver carcinoma cell line (HepG2) as summarized below. Tumor necrosis factor-α (TNF-α), a proinflammatory cytokine, has been shown to activate NF-κB signaling while also activating the c-Jun NH(2)-terminal kinase (JNK) signaling pathway. Other studies have reported that JNK signaling can up-regulate CYP4F11 expression. The results of this study demonstrate that in the presence of TNF-α and the specific NF-κB translocation inhibitor N-[3,5-bis(trifluoromethyl)phenyl]-5-chloro-2-hydroxybenzamide (IMD-0354), there is a greater increase in CYP4F11 expression than that elicited by TNF-α alone, indicating that NF-κB plays an inhibitory role. Moreover, NF-κB stimulation by overexpression of mitogen-activated protein kinase kinase kinase inhibited CYP4F11 promoter expression. CYP4F11 promoter inhibition can also be rescued in the presence of TNF-α when p65, a NF-κB protein, is knocked down. Thus, NF-κB signaling pathways negatively regulate the CYP4F11 gene.
