7124

TNF

DIF|TNF-alpha|TNFA|TNFSF2;tumor necrosis factor;TNF;tumor necrosis factor

APC1 protein|TNF superfamily, member 2|TNF, macrophage-derived|TNF, monocyte-derived|TNF-a|cachectin|tumor necrosis factor alpha|tumor necrosis factor ligand superfamily member 2|tumor necrosis factor-alpha

6p21.3

protein-coding

BACKGROUND: Circulating levels of tumor necrosis factor (TNF) and lymphotoxin-alpha (LT-alpha) have been associated with outcome in solid and hematologic malignancies. Within the TNF gene and the LT-alpha gene, polymorphisms have been identified at nucleotide positions -308 and +252, respectively. The variant alleles for TNF are designated TNF1 and TNF2, the onesfor LT-alpha LT-alpha (10.5 kb) and LT-alpha (5.5 kb). Of interest, TNF2 and LT-alpha (5.5 kb) were shown to be associated with higher TNF and LT-alpha plasma levels than their counterparts. In the present study, we investigated the associations of the above mentioned polymorphisms with risk of relapse in childhood acute lymphoblastic leukemia (ALL) treated according to Berlin-Frankfurt-Munster (BFM) protocols. METHODS: Matched case-control study of64 relapsed and 64 successfully treated non-relapsed childhood B-cell precursor ALL patients of standard and intermediate risk for treatment failure. RESULTS: The odds ratio (OR) for the combined category of TNF1/TNF2 and TNF2/TNF2 genotypes in comparison to the TNF1/TNF1 genotype was 1.17 (95 % confidence interval (CI) = 0.53 - 2.56, P = 0.697). The ORs for the LT-alpha (10.5 kb/5.5 kb) and the LT-alpha (5.5 kb/5.5 kb) genotypes with reference to the LT-alpha (10.5 kb/10.5 kb) genotype were 2.17 (95 % CI = 0.84 - 5.58, P = 0.107) and 0.5 (95 % CI = 0.09 - 2.66, P = 0.418), respectively. CONCLUSIONS: Our results do not suggest a major role of the investigated genetic polymorphisms with regard to risk of relapse in standard- and intermediate-risk childhood B-cell precursor ALL treated according to BFM protocols.

Retinoblastoma is the most common primary intra-ocular tumor in childhood. It has been established that recurrent cytogenetic abnormalities are the hallmark of mostly ALL malignant tumors. Recurrent atypical cytogenetic abnormalities of theshort arm of chromosome 6 were reported in several cases of retinoblastoma, mainly in association with reciprocal translocations. In the present study, alterations of chromosome 6 associated with retinoblastoma were studies in the Y-79 and the WERI-Rb-1 cell lines established from highly malignant retinoblastomas. Fluorescence in situ hybridization (FISH) technique was used toidentify the chromosomal breakpoint on 6p in this cell line. To perform this detection, yeast artificial chromosome (YAC) clones from p21 to p22 bands were used and the breakpoint was localized on 6p21.3. Previous studies had suggested that activation of some genes on 6p could be dependent on a translocation mechanism. expression of a candidate gene localized near the chromosomal breakpoint was measured by immunocytochemistry and flow cytometry techniques. Anenhancement of the tumor necrosis factor-alpha (TNF-alpha) protein expression inY79 cells was detected by this approach. The relationship between TNF-alpha over-expression and the malignancy of retinoblastoma is discussed.

Juvenile myelomonocytic leukemia (JMML) is an aggressive childhood disorder withfew therapeutic options. Granulocyte-macrophage colony-stimulating factor (GM-CSF) and tumor necrosis factor-alpha (TNF-alpha) promote JMML cell growth. Ahyperactive function of the ras oncogene is a hallmark of JMML. We therefore targeted the protein kinase Raf-1 downstream of Ras using a DNA enzyme that degrades mRNA-Raf-1. Western blots of JMML cell lysates revealed phosphorylated Raf-1 protein, indicating constitutive activation. Addition of GM-CSF, but not TNF-alpha, increased phosphorylation of both Raf-1 and the mitogen-activated protein kinases (MAPKs) JNK-1 and ERK-1. Depletion of Raf-1 protein markedly impaired activation of MAPKs, induced substantial inhibition of JMML cell colonyformation, and virtually abolished GM-CSF hypersensitivity in JMML cells. Exogenous TNF-alpha, but not GM-CSF, restored colony formation of JMML cells pretreated with the enzyme. We could not detect any effect of the enzyme on the proliferation of normal bone marrow cells, indicating its specificity and potential safety. When immunodeficient mice engrafted with JMML cells were treated continuously with the enzyme via a peritoneal osmotic mini-pump for 4 weeks, a profound reduction in the JMML cell numbers in the recipient murine bone marrows was found. We conclude that GM-CSF is a chief regulator of JMML growth and exerts its proleukemic effects primarily via the Ras/Raf-1 signaling cascade. TNF-alpha plays a permissive role, being dependent upon GM-CSF to induce JMML cell proliferation. The DNA enzyme efficiently catabolized mRNA-Raf-1 with subsequent inhibition of JMML cell growth, suggesting its potential as a mechanism-based therapy in this fatal leukemia.

Ewing's sarcoma is the second most common human bone tumor in childhood. Here, we investigated the sensitivity of the Ewing tumor cell line, SK-N-MC, to the apoptotic effect of type I (IFNalpha) and type II (IFNgamma) interferons and TNFalpha. We demonstrate that although IFNalpha and TNFalpha alone are unable toinduce cell death, they act in synergy with IFNgamma to induce SK-N-MC cell apoptosis. The synergistic induction of apoptosis correlated with the synergistic induction of TNFalpha-related apoptosis-inducing ligand (TRAIL) mRNA and TRAIL protein synthesis as well as of TRAIL secretion. Preparations of inducer-free supernatants from SK-N-MC cells stimulated with combinations of cytokines were shown to be cytotoxic for untreated SK-N-MC cells. This cytotoxicity was partially inhibited by addition of TRAILR2/Fc fusion protein, indicating that the secreted TRAIL mediates, at least in part, the apoptotic effect displayed by thesupernatants of stimulated SK-N-MC cells. We have shown that the presence of IFNgamma is required to allow the sustained expression of IRF1 in SK-N-MC cells stimulated by addition of IFNalpha or TNFalpha suggesting that IRF1 plays a rolein the synergistic induction of apoptosis by combinations of cytokines. Furthermore, we have shown that inhibition of NF-kappaB activation contributes to the IFNgamma-mediated sensitization to the apoptotic effect of TNFalpha. To our knowledge, this is the first report showing that interferon/cytokine combinations are able to induce TRAIL gene expression and TRAIL protein synthesis and secretion in Ewing sarcoma-derived cells. We believe that the observations reported here might contribute to the development of alternative new approaches to the treatment of Ewing tumors resistant to conventional therapy.#CI- Copyright 2004 Nature Publishing Group

Excessive production of tumor necrosis factor alpha (TNFalpha) may influence therisk and/or progression of hematologic malignancies and has been associated withfebrile episodes at diagnosis. We have examined the putative association of the C-850T polymorphism of the human TNFalpha gene with childhood acute lymphoblastic leukemia (ALL) and with clinical and laboratory findings at diagnosis, in 58 childhood ALL patients from northern Greece. No statistically significant associations have emerged between this polymorphism and either the risk for childhood ALL or presence of fever, anemia, leukocytosis and leukopenia at diagnosis, in this study.

OBJECTIVE: tumor necrosis factor (TNF)-alpha plays a role in the inflammatory process in Crohn disease, a disease with an apparent polygenic basis. We investigated whether polymorphisms in multiple genes involved in the lipopolysaccharide-TNF inflammatory pathway are independently associated with Crohn disease in the Jewish Ashkenazi population. Polymorphisms in CD14, Toll-like receptor 4 (TLR4), and TNF-alpha were studied. In addition, we investigated polymorphisms in the TNF-alpha converting enzyme (TACE) gene, whichto date has not been studied for an association with Crohn disease. PATIENTS ANDMETHODS: To examine whether TLR4 Asp299Gly, CD14-260C/T, TNF-1031T/C, TNF-863C/A, TNF-857C/T, TACE-172C/T, and TACE-154C/A polymorphisms are associated with Crohndisease in the Ashkenazi Jewish population, we analyzed families with at least 1child with Crohn disease for association with these mutations using a family-based association test (transmission disequilibrium test) for analysis. RESULTS: The allelic frequency in the patient population of TLR4 G allele was 8.0%, CD14 T allele was 51.3%, TNF-1031C was 18.8%, TNF-863A was 14.2%, TNF-857Twas 25.2%, TACE172T was 20.7%, and TACE154A was 24.5%. The transmission disequilibrium test transmitted:untransmitted (T:U) result for TLR4G was T:U = 32:20, for CD14T was T:U = 103:88, for TNF-1031C was T:U = 48:56, for TNF-863A was T:U = 39:42, for TNF-857T was T:U = 63:62, for TACE-172C/T was T:U = 48:59, and for TACE-154C/A was T:U = 52:55. No statistically significant associations were observed. CONCLUSIONS: The transmission disequilibrium test did not demonstrate preferential transmission of these variants in Jewish Ashkenazi patients with Crohn disease. These results suggest that these polymorphisms in the TNF/lipopolysaccharide pathway play little or no role in susceptibility to Crohn disease in the Jewish Ashkenazi population.

BACKGROUND: Iron deficiency and the Q248H mutation in the gene, SLC40A1, that encodes for the cellular iron exporter, ferroportin, are both common in African children. The iron status of macrophages influences the pro-inflammatory response of these cells. We hypothesized that Q248H mutation may modify the inflammatory response by influencing iron levels within macrophages. METHODS: The Q248H mutation and circulating concentrations of ferritin, C-reactive protein and selected pro-inflammatory cytokines (interleukin-12, interferon-gamma, TNF-alpha, and macrophage migration inhibitory factor) and anti-inflammatory cytokines (interleukin-4 and interleukin-10) were measured in 69 pre-school children recruited from well-child clinics in Harare, Zimbabwe. RESULTS: In multivariate analysis, both ferroportin Q248H and ferritin <10ug/L were associated with significantly lower circulating concentrations of tumor necrosis factor-alpha. Ferroportin Q248H but not low iron stores was associated with lower circulating macrophage migration inhibitory factor as well. Anti-inflammatory cytokine levels were not significantly associated with either ferroportin Q248H or iron status. CONCLUSIONS: Ferroportin Q248H and low iron stores are both associated with lower circulating tumor necrosis factor-alpha, while only ferroportin Q248H is associated with lower circulating macrophage migration inhibitory factor. Whether the reduced production of tumor necrosis factor-alpha observed in ferroportin Q248H heterozygotes may be of significance in anemia of chronic disease is yet to be determined.#CI- Copyright 2010 Elsevier B.V. ALL rights reserved.

BACKGROUND: Impaired apoptosis has been implicated in the development of childhood adrenocortical tumors (ACT), although the expression of apoptosis-related gene expression in such tumors has not been reported. METHODS:The mRNA expression levels of the genes CASP3, CASP8, CASP9, FAS, TNF, NFKB, andBCL2 were analyzed by quantitative real-time PCR in consecutive tumor samples obtained at diagnosis from 60 children with a diagnosis of ACT and in 11 non-neoplastic adrenal samples. BCL2 and TNF protein expression was analyzed by immunohistochemistry. RESULTS: A significant association was observed between tumor size >/=100 g and lower expression levels of the BCL2 (P=0.03) and TNF (P=0.05) genes; between stage IV and lower expression levels of CASP3 (P=0.008),CASP9 (P=0.02), BCL2 (P=0.002), TNF (P=0.05), and NFKB (P=0.03); Weiss score >/=3 and lower expression of TNF (P=0.01); unfavorable event and higher expression values of CASP9 (P=0.01) and lower values of TNF (P=0.02); and death and lower expression of BCL2 (P=0.04). Underexpression of TNF was associated with lower event-free survival in uni- and multivariate analyses (P<0.01). Similar results were observed when patients with Weiss score <3 were excluded. CONCLUSION: This study supports the participation of apoptosis-related genes in the biology and prognosis of childhood ACT and suggests the complex role of these genes in the pathogenesis of this tumor.

BACKGROUND: It has been suggested that fetal carcinogenic exposure might lead topredisposition to develop cancer during childhood or in later life possibly through modulation of the fetal transcriptome. Because gender effects in the incidence of childhood cancers have been described, we hypothesized differences at the transcriptomic level in cord blood between male and female newborns as a consequence of fetal carcinogenic exposure. The objective was to investigate whether transcriptomic responses to dietary genotoxic and nongenotoxic carcinogens show gender-specific mechanisms-of-action relevant for chemical carcinogenesis. METHODS: Global gene expression was applied in umbilical cord blood samples, the CALUX-assay was used for measuring dioxin(-like), androgen(-like), and estrogen(-like) internal exposure, and acrylamide-hemoglobin adduct levels were determined by mass spectrometry adduct-FIRE-procedure(TM). Tolink gene expression to an established phenotypic biomarker of cancer risk, micronuclei frequencies were investigated. RESULTS: While exposure levels did not differ between sexes at birth, important gender-specific differences were observed in gene expressions associated with these exposures linked with cell cycle, the immune system and more general cellular processes such as posttranslation. Moreover, oppositely correlating leukemia/lymphoma genes between male and female newborns were identified in relation to the different biomarkersof exposure that might be relevant to male-specific predisposition to develop these cancers in childhood. CONCLUSIONS/IMPACT: This study reveals different transcriptomic responses to environmental carcinogens between the sexes. In particular, male-specific TNF-alpha-NF-kB signaling upon dioxin exposure and activation of the Wnt-pathway in boys upon acrylamide exposure might represent possible mechanistic explanations for gender specificity in the incidence of childhood leukemia.#CI- 2012 AACR

BACKGROUND: Febrile convulsions (FCs), occurring between 6 months and 6 years ofage is the most common seizure disorder during childhood. The febrile response is thought to be mediated by the release of pyrogenic cytokines, such as tumor necrosis factor and interleukin-1 (IL-1). There is a significant relationship between genetic components for susceptibility of FCs and different report mutation. We investigated association between two polymorphisms in the tumor necrosis factor (TNF)-alpha promoter region (G-308A, C-850T) and FCs in the southwest area of Iran. MATERIALS AND METHODS: In this matched case-control study, 100 patients with febrile convulsion as case group and 130 healthy children as control group were enrolled in the study. Peripheral blood samples were collected and DNA was extracted by standard phenol-chloroform method. The genotype and allele frequencies of TNF- alpha polymorphisms in case and control groups were determined by using PCR-RFLP (polymerase chain reaction restriction fragment length polymorphism) method. Statistical analysis was done using Chi-square test. RESULTS: The average age of case and control groups were 3.4 +/- 1.4 and 3.4 +/- 1.2 years, respectively. There was no significant difference between age and sex in both the groups (P > 0.05). A family history of febrile convulsion was detected in 44% of patients. Moreover, the simple febrile convulsion was detected in 85% of the case group. CONCLUSION: RFLP analysis of TNF- alpha promoter region polymorphisms, considering P = 0.146 and P = 0.084 for G-308A and C-850T, respectively, showed no correlation between TNF- alpha polymorphisms and predisposition to simple febrile, based on the kind of convulsion (atypical and simple febrile convulsion). We found a significant relation between genotype distribution of G-308A and atypical febrile convulsionin case group (P = 0.04). A significant correlation between genotype distribution of G-308A and atypical febrile convulsion in the case group was found, but therewas no correlation between TNF- alpha polymorphisms at positions of -308A, and 850T and predisposition to simple febrile convulsion. Further studies are neededto understand clinical usefulness of this correlation.

Retinoblastoma is an intraocular malignancy of childhood, which is very radiosensitive. gamma-irradiation, however, induces side effects, and the precise mechanisms of tumor cell death after the treatment remain unknown. In this study, we demonstrated that gamma-irradiation induced apoptosis (programmed cell death)in human retinoblastoma cell lines Y79 and WERI-Rb-1. The expression levels of p53, tumor suppressor gene product, and its-associated protein, WAF1/CIP1, were both up-regulated, and function of p53 was remarkably activated after the treatment. Moreover, apoptosis was induced in the absence of gamma-irradiation by overexpression of the WAF1/CIP1 gene in both retinoblastoma cells. These resultsindicate that the transfer of the WAF1/CIP1 gene may have potential for the treatment of human retinoblastomas instead of gamma-irradiation.