The Effect of Socioeconomic Status and Race/Ethnicity on the Risk of Presenting With Advanced Stage at Diagnosis in Embryonal Tumors.

We evaluated whether socioeconomic status (SES), race/ethnicity, and their interaction were associated with the presentation of advanced stage at diagnosis in embryonal tumors. Children 0 to 19 years of age diagnosed with embryonal tumors between 2006 and 2018 were identified from the US Surveillance, Epidemiology, and End Results program database specialized with Census Tract SES/Rurality. SES quintile was derived from a composite index for census tracts. We performed logistic regression to estimate odds ratios (ORs) and 95% confidence intervals by SES and race/ethnicity, adjusting for sex, age, and diagnosis year. Overall, no significant associations were found between either SES or race/ethnicity and the risk of presenting with advanced stage at diagnosis, although patterns of risk reductions were observed in atypical teratoid/rhabdoid tumors and embryonal rhabdomyosarcoma with increasing SES. In the stratified analysis, decreased odds of presenting with advanced-stage embryonal rhabdomyosarcoma were observed for Hispanics with higher SES (OR: 0.24, 95% Confidence Interval: 0.08-0.75) compared with Hispanics with lower SES. Future studies incorporating individual-level SES, cancer-specific staging information, and potential demographic, clinical, epidemiological, and genetic risk factors are warranted to confirm our findings.

The association between maternal nutrient intake during pregnancy and the risk of sporadic unilateral retinoblastoma among offspring.

Previous studies have associated maternal diet during pregnancy with the development of sporadic unilateral retinoblastoma (RB), but few studies have focused on the role of individual nutrients. The aim of this study is to investigate the association between maternal nutrient intake during pregnancy and the development of sporadic unilateral RB in the offspring. A modified food frequency questionnaire, with additional questions on supplement use, was completed via a phone interview. Cases were recruited from hospitals and controls were comprised of friends and relatives of the patient without a history of cancer. Overall, 168 sporadic unilateral RB cases and 145 controls were included in case-control study. We performed logistic regression to estimate odds ratios (ORs) and 95 % confidence intervals (CI), adjusting for child's age, child's sex, parental race/ethnicity, maternal education, total calorie intake during pregnancy, maternal age at birth, maternal smoking during pregnancy, pre-pregnancy body mass index, maternal weight gain during pregnancy, paternal age at birth, and maternal multivitamin use in the year before pregnancy. In the adjusted model, the interquartile (IQR) increase in vitamin A intake, which was measured in retinol activity equivalent (RAE; OR: 0.64, 95 % CI: 0.46-0.90), and vitamin D intake (OR: 0.62, 95 % CI: 0.42-0.91) significantly reduced the risk of sporadic unilateral RB. These findings suggest that a higher intake of vitamins A and D can be a protective factor for sporadic unilateral RB. Further analyses in consideration of multi-exposures such as parental occupational exposures are warranted to discover the complex etiology of sporadic unilateral RB. In addition, the role of nutritional epigenetics for how maternal nutrient intake influences the risk of sporadic unilateral RB in the offspring still needs to be explored.

Exploration of genetic ancestry and socioeconomic status in the incidence of neuroblastoma: An ecological study.

Although global differences in the incidence of neuroblastoma have been examined, the underlying mechanism has yet to be elucidated. Previous studies have suggested genetic ancestry and human development index (HDI) as contributing factors, but few studies have been conducted at the international level. Here, we aimed to examine whether the frequency of common genomic variation associated with neuroblastoma can affect its risk at the ecological level with consideration of the HDI. Minor allele frequencies (MAFs) for 22 single-nucleotide polymorphisms (SNPs) were abstracted from the Geography of Genetic Variants Browser. The number of incident neuroblastomas for each population was obtained from the Cancer Incidence in Five Continents series. Further, population pseudo-polygenic risk scores (pp-PRSs) were calculated as a sum of MAFs at the population level, each of which was weighted by effect sizes from prior studies. Negative binomial regression was used to estimate the incidence rate ratios (IRRs) and the 95% confidence intervals (CIs) to examine whether differences in MAFs across the population influence the risk of neuroblastoma, with and without adjustment for HDI and whether pp-PRSs can be a predictor of the risk of neuroblastoma. Overall, our results indicated that the neuroblastoma risk associated with variation in SNP frequency could not be differentiated from that of HDI at the ecological level. Additionally, pp-PRSs were not significantly associated with the risk of neuroblastoma (IRR: 0.99, 95% CI: 0.62-1.60). Further study using individual-level data is warranted to minimize the bias related to the use of population-level data in this study.

Daytime sleepiness among Midwestern firefighters.

Excessive daytime sleepiness (EDS) is the most common symptom of sleep-disorders. Although high levels of EDS have been reported among firefighters, there is a limited understanding on EDS in firefighters. A cross-sectional survey was conducted in full-time firefighters (n = 275). The survey included the Epworth Sleepiness Scale to assess EDS, as well as other questions related to job characteristics, health history, and demographics. Firefighters reported inadequate sleep hours, extremely short sleep latencies, and high EDS higher than the general population. A majority of firefighters indicated a willingness to consult with a sleep professional. Risk factors associated with EDS included at least five years of professional experiences and depression diagnosis. Our findings suggest that EDS is widespread among firefighters. With increasing time on the job, frequencies of both obesity and EDS increase, which can have severe and compounding effects on health and safety.

Association between prenatal exposure to PM2.5 and the increased risk of specified infant mortality in South Korea.

BACKGROUND: Findings from previous studies on the association between exposure to fine particulate matter (PM2.5) and the risk of infant mortality were inconsistent. Thus, two main objectives of our study were to examine the association between exposure to PM2.5 and specified infant mortality and to identify critical trimesters. METHODS: We retrospectively created a birth cohort of singleton full-term infants born in South Korea between 2010 and 2015 using national birth and infant mortality data. The specified causes of infant mortality were circulatory and respiratory diseases, perinatal conditions, congenital anomalies, and sudden infant death syndrome. We performed 1:10 propensity score matching for various exposure windows: each trimester, prenatal, and postnatal (up to age 1). Conditional logistic regression was applied to estimate odds ratios (ORs) and 95% confidence intervals (CIs), while accounting for gestational age, birth weight, maternal education level, season of birth, and regions (metropolitan areas/provinces). We also conducted sex-stratified analyses and used different matching ratios for sensitivity analyses. RESULTS: A total of 2,501,836 births and 761 deaths (0.03%) were identified in the birth cohort. We found an increased risk of infant mortality per 10 µg/m3 increase in PM2.5 exposure during the prenatal period (OR: 1.29, 95% CI: 1.07-1.55). Exposure in the 1st and 2nd trimesters was linked to an elevated risk (OR: 1.19, 95% CI: 1.02-1.37; OR: 1.21, 95% CI: 1.04-1.40). However, no association was shown in the third trimester. PM2.5 exposure in the 1st and 2nd trimesters was associated with elevated male infant mortality, but did not reach statistical significance in female infants. The use of different matching ratios did not significantly affect the results. CONCLUSION: The study findings suggest that exposure to PM2.5 could affect infant mortality differently by the timing of exposure and sex, which suggests a relation to fetal development. However, further investigations are warranted.

Children's Environmental Health Indicators in Context of the Sustainable Development Goals for Small Island Developing States.

The unique environmental vulnerability of small island developing states (SIDS) is likely to impact negatively on children's health. Children's environmental health indicators (CEHI) are standardized measures that can be used to assess the environmental exposures and their resulting health outcomes in children. This study sought to utilize the United Nations (UN) global Sustainable Development Goals (SDGs), with their associated targets and indicators, as a framework for a CEHI proposal for SIDS. Exposure-side indicators were taken from key themes from the 2012 Rio+20 UN Conference on Sustainable Development, and health-side indicators were selected based on the most significant contributors to the burden of disease in children. The multiple-exposures⁻multiple-effect (MEME) framework was then used to show the relationships between environmental exposures and children's health outcomes. The framework was populated with available data from the World Bank's DataBank. Whilst there was some data available at a population level, major gaps in both exposure-side and health-side indicators were revealed. In order to progress children's environmental health in SIDS, a further piece of work is required to propose a fully prioritized set of exposure-side and health-side CEHIs; based on, but not exclusively linked to, the SDGs.

Does exposure to PM10 decrease age at menarche?

BACKGROUND: There has been a consistent decrease in age at menarche in South Korea. A potential risk factor for early menarche is exposure to particulate matter (PM), because endocrine-disrupting compounds emitted into air from anthropogenic sources may be incorporated into PM. The objective of this study was to examine the association between pre-menarcheal exposure to PM ≤ 10 µm in diameter (PM10) and age at menarche in adolescents of South Korea using Korea National Health and Nutrition Examination Survey (KNHANES) 2010-2012 data. METHODS: We used self-reported age at menarche of 639 girls aged 13-17 years in this study. The cut-off age for early menarche was set to 12 years. Based on each subject's address, 1-year, 2-year, and 3-year averages of annual mean PM10 concentrations (models 1-3) were linked to KNHANES. Models were adjusted for body mass index (BMI), city size, household income level, maternal age at menarche, and second-hand smoke exposure at home. SURVEYREG and SURVEYLOGISTIC procedures were used to address the complex survey design of KNHANES. RESULTS: Overall analysis showed that exposure to PM10 has a significant effect on decreasing age at menarche. Multiple linear regression results suggested that each 1 µg/m3 increase in 1-year, 2-year, 3-year averages of annual mean PM10 concentrations accelerated age at menarche by 0.046 years (95% CI: -0.064, -0.027; p < .0001), 0.038 years (95% CI: -0.059, -0.018; p = 0.0003),and 0.031 years (95% CI: -0.047, -0.015; p = 0.0002), respectively. Adjusted ORs for a 1 µg/m3 increase in PM10 concentration were 1.08 (95% CI: 1.04 -1.12) for model 1, 1.06 (95% CI: 1.02 -1.10) for model 2, and 1.05 (95% CI: 1.01 -1.09) for model 3. CONCLUSION: Our findings suggest that elevated PM10 concentration can decrease age at menarche. This is the first study that investigates the association between exposure to PM10 and age at menarche using a nationally representative sample of Koreans.

Children's Environmental Health Indicators for Low- and Middle-Income Countries in Asia.

BACKGROUND: Given that low- and middle-income countries (LMICs) in Asia still have high child mortality rates, improved monitoring using children's environmental health indicators (CEHI) may help reduce preventable deaths by creating healthy environments. OBJECTIVES: Thus, the aim of this study is to build a set of targeted CEHI that can be applied in LMICs in Asia through the CEHI initiative using a common conceptual framework. METHODS: A systematic review was conducted to identify the most frequently used framework for developing CEHI. Due to the limited number of eligible records, a hand search of the reference lists and an extended search of Google Scholar were also performed. Based on our findings, we designed a set of targeted CEHI to address the children's environmental health situation in LMICs in Asia. The Delphi method was then adopted to assess the relevance, appropriateness, and feasibility of the targeted CEHI. FINDINGS: The systematic review indicated that the Driving-Pressure-State-Exposure-Effect-Action framework and the Multiple-Exposures-Multiple-Effects model were the most common conceptual frameworks for developing CEHI. The Multiple-Exposures-Multiple-Effects model was adopted, given that its population of interest is children and its emphasis on the many-to-many relationship. Our review also showed that most of the previous studies covered upper-middle- or high-income countries. The Delphi results validated the targeted CEHI. The targeted CEHI were further specified by age group, gender, and place of residence (urban/rural) to enhance measurability. CONCLUSIONS: Improved monitoring systems of children's environmental health using the targeted CEHI may mitigate the data gap and enhance the quality of data in LMICs in Asia. Furthermore, critical information on the complex interaction between the environment and children's health using the CEHI will help establish a regional environmental children's health action plan, named "The Children's Environment and Health Action Plan for Asia."

NFIB is a potential target for estrogen receptor-negative breast cancers.

BACKGROUND: The association between nuclear factor I/B (NFIB) gene and triple negative breast cancer has been previously suggested. METHODS: We investigated the relationship between NFIB mRNA and protein expression and molecular subtypes of breast cancer as well as the effect of NFIB silencing on the proliferation and apoptosis of breast cancer cells. Also, the clinical importance of NFIB expression was investigated in 163 breast cancer patients. RESULTS: By using 20 frozen human breast cancer tissues and various breast cancer cell lines, we observed a significant high level of NFIB mRNA level in triple negative breast cancer. NFIB protein was upregulated in ER negative breast cancer tissues but the expression level was similar between HER2 subtype and triple negative subtype. The clinical significance of NFIB was further examined in a tissue microarray from 163 invasive breast cancer patients, and the immunohistochemistry results showed a significant association between NFIB expression and nuclear grade, ER, and HER2 expression status. NFIB positive tumors were more likely to have high nuclear grade, ER negativity and HER2 over-expression. HCC1954 cells transfected with siRNA against NFIB showed a significant reduction in cell proliferation and increase in apoptotic signaling pathway. CONCLUSIONS: Our results show a potential role of NFIB as a novel target in ER negative breast cancers.

Evidences for correlation between the reduced VCAM-1 expression and hyaluronan synthesis during cellular senescence of human mesenchymal stem cells.

Mesenchymal stem cells (MSCs) undergo cellular senescence during in vitro expansion culture, which accompanies the loss of migration and homing abilities. In this study, we analyzed expression levels of several surface markers of human MSCs at different passages of expansion culture. It has been shown that expression of vascular cell adhesion molecule-1 (VCAM-1) was most markedly decreased among the tested markers in the senescent MSCs. Interestingly the reduced VCAM-1 expression could be restored by applying hyaluronan, a major glycosaminoglycan ligand of CD44, to the culture. It was found that the hyaluronan level in extracellular and pericellular matrices was greatly reduced in the senescent MSCs, mainly due to the decreased expression of hyaluronan synthases, suggesting a correlation between the reduced VCAM-1 expression and hyaluronan synthesis. In fact, when hyaluronan synthases were knock-downed by siRNA transfection, the VCAM-1 expression was also reduced. Our results indicate that VCAM-1 expression in the senescent MSCs was down-regulated because of the reduced synthesis of hyaluronan. Thus, we suggest that hyaluronan supplementation in expansion culture of MSCs would compensate adverse effects induced by its decreased synthesis and subsequently enhance cell adhesion and migration abilities.

DNA copy number alterations and expression of relevant genes in triple-negative breast cancer.

Triple-negative breast cancer (TNBC) is defined by a lack of expression of estrogen, progesterone, and HER2 receptors, and genetically most of them fall into the basal subgroup of breast cancer. The important issue of TNBC is poorer clinical outcome and absence of effective targeted therapy. In this study, we sought to identify DNA copy number alterations and expression of relevant genes characteristic of TNBC to discover potential therapeutic targets. Frozen tissues from 114 breast cancers were analyzed using high-resolution array comparative genomic hybridization. The classification into subtype was determined by estrogen and progesterone receptor expression, and by the presence or absence of gain on the ERBB2 containing clone. The ACE algorithm was used for calling gain and loss of clones. Twenty-eight cases (25%) were classified as TNBC. Recurrent gains (> or =25%) unique to TNBC were 9p24-p21, 10p15-p13, 12p13, 13q31-q34, 18q12, 18q21-q23, and 21q22. Two published gene expression array data sets comparing basal subtype versus other subtype breast cancers were used for searching candidate genes. Of the genes upregulated in the basal subtype, 45 of 686 genes in one data set and 59 of 1,428 in the second data set were found to be located in the gained regions. Of these candidate genes, gain of NFIB (9p24.1) was specific for TNBC in a validation set by real-time PCR. In conclusion, we have identified recurrently gained regions characteristic of TNBC, and found that NFIB copy number and expression is increased in TNBC across the data sets. This article contains Supplementary Material available at http://www.interscience.wiley.com/jpages/1045-2257/suppmat.

Rosiglitazone promotes tumor necrosis factor-related apoptosis-inducing ligand-induced apoptosis by reactive oxygen species-mediated up-regulation of death receptor 5 and down-regulation of c-FLIP.

Death receptor 5 (DR5/TRAIL-R2) is an apoptosis-inducing membrane receptor for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). In this study, we show that rosiglitazone sensitizes human renal cancer cells to TRAIL-mediated apoptosis, but not normal human mesangial cells. Furthermore, because rosiglitazone-enhanced TRAIL-mediated apoptosis is induced in various types of cancer cells but is not interrupted by Bcl-2 overexpression, this combinatory treatment may provide an attractive strategy for cancer treatment. We found that treatment with rosiglitazone significantly induces DR5 expression at both its mRNA and its protein levels, accompanying the generation of reactive oxygen species (ROS). Both treatment with DR5/Fc chimeric protein and silencing of DR5 expression using small interfering RNAs attenuated rosiglitazone plus TRAIL-induced apoptosis, showing the critical role of DR5 in this cell death. Pretreatment with GSH significantly inhibited rosiglitazone-induced DR5 up-regulation and the cell death induced by the combined treatment with rosiglitazone and TRAIL, suggesting that ROS mediate rosiglitazone-induced DR5 up-regulation, contributing to TRAIL-mediated apoptosis. However, both DR5 up-regulation and sensitization of TRAIL-mediated apoptosis induced by rosiglitazone are likely PPARgamma-independent, because a dominant-negative mutant of PPARgamma and a potent PPARgamma inhibitor, GW9662, failed to block DR5 induction and apoptosis. Interestingly, we also found that rosiglitazone treatment induced down-regulation of cellular FLICE-inhibitory protein (c-FLIPs), and ectopic expression of c-FLIPs attenuated rosiglitazone plus TRAIL-mediated apoptosis, demonstrating the involvement of c-FLIPs in this apoptosis. Taken together, the results of this study demonstrate that rosiglitazone enhances TRAIL-induced apoptosis in various cancer cells by ROS-mediated DR5 up-regulation and down-regulation of c-FLIPs.

The novel phospholipase C activator, m-3M3FBS, induces apoptosis in tumor cells through caspase activation, down-regulation of XIAP and intracellular calcium signaling.

We investigated the effect of the novel phospholipase C activator, m-3M3FBS, on the apoptosis of human renal Caki cancer cells. Treatment with m-3M3FBS induced apoptosis of Caki cells, which was accompanied by accumulation of sub-G1 phase and DNA fragmentation. We found that induction of apoptosis is a common response of several cancer cell types to m-3M3FBS treatment. Overexpression of Bcl-2 and c-FLIPs fails to block m-3M3FBS-induced apoptosis. However, ectopic expression of XIAP partly inhibits m-3M3FBS-induced apoptosis in Caki cells. m-3M3FBS-induced apoptosis appeared to involve the XIAP down-regulation and caspase activation. m-3M3FBS also induced the expression of a potential proapoptotic gene, C/EBP homologous protein (CHOP), however, suppression of CHOP expression by small interfering RNA did not abrogate the m-3M3FBS-induced apoptosis. In addition, inhibition of phospholipase C (PLC) or chelation of intracellular calcium prevented m-3M3FBS-induced apoptosis in Caki cells, suggesting that the involvement of PLC pathway and intracellular calcium signaling on the apoptosis in m-3M3FBS-treated Caki cells. Collectively, our present results suggest that m-3M3FBS-induced apoptosis in Caki cells may result from the activation of caspase, down-regulation of XIAP and intracellular Ca(2+) release pathway and that m-3M3FBS treatment might overcome the anti-apoptotic effect of Bcl-2 or c-FLIPs in cancer cells.

Mithramycin A sensitizes cancer cells to TRAIL-mediated apoptosis by down-regulation of XIAP gene promoter through Sp1 sites.

Mithramycin A is a DNA-binding antitumor agent, which has been clinically used in the therapies of several types of cancer and Paget's disease. In this study, we investigated the combined effect of mithramycin A and tumor necrosis factor-alpha-related apoptosis-inducing ligand (TRAIL) on apoptosis of cancer cells. In Caki renal cancer cells, which are resistant to TRAIL, cotreatment with subtoxic doses of mithramycin A and TRAIL resulted in a marked increase in apoptosis. This combined treatment was also cytotoxic to Caki cells overexpressing Bcl-2 but not to normal mesengial cells. Moreover, apoptosis by the combined treatment with mithramycin A and TRAIL was dramatically induced in various cancer cell types, thus offering an attractive strategy for safely treating malignant tumors. Mithramycin A-stimulated TRAIL-induced apoptosis was blocked by pretreatment with the broad caspase inhibitor zVAD-fmk or Crm-A overexpression, showing its dependence on caspases. We found that mithramycin A selectively down-regulated XIAP protein levels in various cancer cells. Luciferase reporter assay and the chromatin immunoprecipitation assay using the XIAP promoter constructs show that mithramycin A down-regulates the transcription of XIAP gene through inhibition of Sp1 binding to its promoter. Although XIAP overexpression significantly attenuated apoptosis induced by mithramycin A plus TRAIL, suppression of XIAP expression by transfection with its small interfering RNA prominently enhanced TRAIL-induced apoptosis. We present here for the first time that mithramycin A-induced suppression of XIAP transcription plays a critical role in the recovery of TRAIL sensitivity in various cancer cells.

Caspase-dependent and caspase-independent apoptosis induced by evodiamine in human leukemic U937 cells.

Evodiamine is one of the major bioactive compounds that have been isolated and purified from the fruit of Evodiae fructus. Evodiamine exhibits antitumor activities against the human tumor cells, including multidrug-resistant tumor cells. However, the molecular mechanism involved in cell death induced by evodiamine treatment remains poorly understood. In the present study, we showed that evodiamine activated the caspase-dependent apoptotic pathway. This apoptosis was only partially inhibited by a pancaspase inhibitor benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone, which suggested that evodiamine-induced apoptosis in leukemic U937 cells is partially caspase independent. We observed the nuclear translocation of apoptosis-inducing factor in evodiamine-induced apoptosis of U937 cells, which may be responsible for the caspase-independent apoptotic execution. We next showed that evodiamine induced the substantial amount of apoptosis both in Bcl-2- and Akt-overexpressing U937 cells but not in human peripheral blood mononuclear cells. Although benzyloxycarbonyl-Val-Ala-Asp-fluoromethyl ketone inhibited caspase activity in Bcl-2-overexpressing U937 cells, it completely prevented neither the induction of apoptosis or the nuclear translocation of apoptosis-inducing factor, which suggests that evodiamine is, at least in part, able to bypass the resistance of leukemia cells via caspase-independent apoptotic pathways. Thus, therapeutic strategy using evodiamine may warrant further evaluation.

Curcumin sensitizes tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-mediated apoptosis through CHOP-independent DR5 upregulation.

Death receptor DR5 (DR5/TRAIL-R2) is an apoptosis-inducing membrane receptor for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL). In this study, we showed that curcumin, a plant product containing the phenolic phytochemical, is a potent enhancer of TRAIL-induced apoptosis through upregulation of DR5 expression. Both treatment with DR5/Fc chimeric protein and silencing of DR5 expression using small interfering RNA (siRNA) attenuated curcumin plus TRAIL-induced apoptosis, showing that the critical role of DR5 in this cell death. Curcumin also induced the expression of a potential pro-apoptotic gene, C/EBP homologous protein (CHOP), both at its mRNA and protein levels. However, suppression of CHOP expression by small interfering RNA did not abrogate the curcumin-mediated induction of DR5 and the cell death induced by curcumin plus TRAIL, demonstrating that CHOP is not involved in curcumin-induced DR5 upregulation. Taken together, the present study demonstrates that curcumin enhances TRAIL-induced apoptosis by CHOP-independent upregulation of DR5.

Curcumin sensitizes tumor necrosis factor-related apoptosis-inducing ligand (TRAIL)-induced apoptosis through reactive oxygen species-mediated upregulation of death receptor 5 (DR5).

Curcumin exhibits anti-inflammatory and antitumor activities. Although its functional mechanism has not been elucidated so far, numerous studies have shown that curcumin induces apoptosis in cancer cells. In the present study, we show that subtoxic concentrations of curcumin sensitize human renal cancer cells to the tumor necrosis factor-related apoptosis inducing ligand (TRAIL)-mediated apoptosis. This apoptosis induced by the combination of curcumin and TRAIL is not interrupted by Bcl-2 overexpression. We found that treatment with curcumin significantly induces death receptor 5 (DR5) expression both at its mRNA and protein levels, accompanying the generation of the reactive oxygen species (ROS). Not only the pretreatment with N-acetylcystine but also the ectopic expression of peroxiredoxin II, an antioxidative protein, dramatically inhibited the apoptosis induced by curcumin and TRAIL in combination, blocking the curcumin-mediated DR5 upregulation. Taken together, the present study demonstrates that curcumin enhances TRAIL-induced apoptosis by ROS-mediated DR5 upregulation.

8-Hydroxyquinoline inhibits iNOS expression and nitric oxide production by down-regulating LPS-induced activity of NF-kappaB and C/EBPbeta in Raw 264.7 cells.

In activated macrophage, large amounts of nitric oxide (NO) are generated by inducible nitric oxide synthase (iNOS), resulting in acute or chronic inflammatory disorders. In Raw 264.7 cells stimulated with lipopolysaccharide (LPS) to mimic inflammation, 8-hydroxyquinoline (8HQ) inhibited the LPS-induced expression of both iNOS protein and mRNA in a parallel dose-dependent manner. 8HQ did not enhance the degradation of iNOS mRNA. To investigate the mechanism by which 8HQ inhibits iNOS gene expression, we examined the activation of MAP kinases in Raw 264.7 cells. We did not observe any significant change in the phosphorylation of MAPKs between LPS alone and LPS plus 8HQ-treated cells. Moreover, 8HQ significantly inhibited the DNA-binding activity of nuclear factor-kappaB (NF-kappaB) and CCAAT/enhancer-binding protein beta (C/EBPbeta), but not activator protein-1 and cAMP response element-binding protein. Taken together, these results suggest that 8HQ acts to inhibit inflammation through inhibition of NO production and iNOS expression through blockade of C/EBPbeta DNA-binding activity and NF-kappaB activation.