PURPOSE: To evaluate the expression of senescence-related proteins according to the aging process and to determine the role of senescence-related proteins in the bone tissue and their effects on the process of bone union. MATERIALS AND METHODS: 18 Sprague-Dawley rats (8 weeks old: 7, 32 weeks old: 6, and 70 weeks old: 5) were used in the experiment. A unilateral closed femur fracture was made, and the fracture callus was obtained 2 weeks after the fracture. The ossification process was observed in proliferative chondrocytes, the hypertrophic chondrocytes, and in the mesenchymal layer individually by immunohistochemistry, using p16, p21, c-fos and c-jun antibodies. RESULTS: There was no significant differences in the manifestation of p-16, p-21, c-fos, c-jun gene according to the age. The positive ratio of p-16 was maximal in proliferative chondrocytes (54.93%) and decreased in the mesenchymal layer (46.48%), and in hypertrophic chondrocytes (10.85%), in order. The positive ratio of c-fos was maximal in proliferative chondrocytes (73.32%) and decreased in the mesenchymal layer (51.84%), and in hypertrophic chondrocytes (9.64%), in order. CONCLUSION: We believe that senescent genes in the bone tissue participate in the differentiation of osteochondral cells and in the process of fracture callus ossification.
Aging ; Animals ; Antibodies ; Bone and Bones ; Bony Callus* ; Chondrocytes ; Femur ; Genes, jun ; Immunohistochemistry ; Rats* ; Rats, Sprague-Dawley

AIMTo investigate the effects of anoxia/reoxygenation on Fos and Jun expression and apoptosis in cultured rat hippocampal neurons.

METHODSThe hippocampal neurons cultured for 12 d were exposed to anoxia environment (90% N2 + 10% CO2) for 4 h and then reoxygenated for 24 h and 72 h. The neurons were immunocytochemically stained using the antiserum against Fos and Jun, and the apoptosis were detected by using the terminal deoxynucleotidyl transferase mediated biotinylated deoxyuridine triphosphate nickel end labeling (TUNEL) method and flow cytometric analysis.

RESULTSThe percentage of Fos and Jun positive neurons and apoptosis neurons in cultured hippocampal neurons after anoxia/reoxygenation increased than those in control.

CONCLUSIONThe occurrence of neurons apoptosis is related to the increase in Fos and Jun expression in cultured hippocampal neurons after anoxia/reoxygenation.

Animals ; Apoptosis ; Cell Hypoxia ; Cells, Cultured ; Genes, fos ; Genes, jun ; Hippocampus ; metabolism ; Neurons ; metabolism ; Oxygen ; metabolism ; Rats ; Rats, Wistar

PURPOSE: Fragile sites are points on chromosomes which tend to break non-randomly when exposed to specific chemical agents or conditions of tissue culture. The chromosomal break induced by the antineoplastic drug, 1-beta-D-arabinofuranosyl-cytosine(Ara-c), was investigated to study the laboratory conditions in which the incidence of chromosomal break could be enhanced. Besides, the fragile sites induced by Ara-C were investigated and compared to the already known locations of the specific chromosomal alterations observed in specific neoplasms. METHODS: T-lymphocytes from theree normal males and three females were cultured for 48 hours. Cells from each individual were exposed to the Ara-C for an additional 24 hours. After the caffeine was added during the last six hours culture, the metaphase chromosomes were prepared following the conventional method. A site was considered fragile if it was found to break two or more per 100 chromosomal breaks in more than four of six individuals tested. RESULTS: Ara-C induced 252.1 chromosomal breaks per 100 mitotic cells and this result was significantly higher than that of the control, which induced 25.2 breaks(P<0.05). The incidence of the chromosomal break by Ara-C was higher, if cultured in the MEM-FA, which has no folic acid, than in the RPMI 1640 which contains enough folic acid(P<0.05). The most common break site by Ara-C was 3p14.2(FRA3B). There were 20 fragile sites induced by Ara-C. Among these 20 fragile sites, seven coincided with the locations of the mapped oncogenes, JUN, SKI, REL, N-MYC, FHIT, MET, ETS-1, and FOS. CONCLUSIONS: S phase specific chemotherapeutic agent, Ara-C, induced the expression of the chromosomal fragile sites effectively using the T-lymphocyte in vitro. Some of the fragile sites by Ara-C highly coincided with the oncogenes and neoplasm specific chromosome breakpoints. In this regard, the fragile sites reported here could provide the unknown neoplasm related chromosomal alternation points.
Caffeine ; Chromosome Breakage* ; Chromosome Breakpoints ; Cytarabine ; Female ; Folic Acid ; Genes, jun ; Humans ; Incidence ; Male ; Metaphase ; Oncogenes ; S Phase ; T-Lymphocytes

AIMTo study the effects of hypoxic preconditioning on anoxic tolerance and Jun expression in cultured rat hippocampal neurons after anoxia/reoxygenation.

METHODS12 day cultured hippocampal neurons in control and hypoxic preconditioning group were exposed to anoxic environment (0.90L/L N2 + 0.10 L/L CO2) for 4 h, and then reoxygenated for either 24 h or 72 h. The neurons were immunocytochemically stained using the antiserum against Jun. The number of survival neurons and the percentage of Jun expressing neurons were investigated.

RESULTSThe percentage of Jun expressing neurons induced by anoxia in hypoxic-preconditioning group was significantly less than that in control group. The number of survival neurons was more in the hypoxic-preconditioning group than that in control group after anoxic reoxygenation.

CONCLUSIONHypoxic-preconditioning can induce the development of anoxic-tolerance in cultured hippocampal neurons. The decrease in Jun expressing neurons in hippocampus may be an adaptive reaction to acute anoxia.

Animals ; Animals, Newborn ; Cell Hypoxia ; Cells, Cultured ; Genes, jun ; Hippocampus ; metabolism ; Neurons ; metabolism ; Oxygen ; metabolism ; Rats ; Rats, Wistar

Apoptosis ; Genes, fos ; Genes, jun ; Heat-Shock Proteins ; physiology ; Humans ; Liver ; blood supply ; Liver Transplantation ; adverse effects ; NF-kappa B ; metabolism ; Nitric Oxide ; physiology ; Reperfusion Injury ; etiology

The expression of the p53 protein (p53) was compared with those of several oncogenes including c-fos (Fos), c-jun (Jun), and epidermal growth factor receptor (EGFR1) using immunohistochemistry in frozen and paraffin-embedded sections of 25 basal cell carcinomas (BCCs) to find out any correlation between p53 and oncogenes in the pathogenesis of human BCC. In normal skin, positive reactions were obtained for EGFR1 and Fos, while p53 and Jun were negative in all cases. In the lesions, EGFR1 was observed in all cases and p53 was positive in 9 of 25 (36%). Fos was expressed in 21 of 25 (84%) and four negative cases were all p53-positive; this negative correlation between p53 and Fos staining was statistically significant (P< 0.01). Jun was detected in 14 of 20 (70%) and no significant relationship was observed between the expression of Jun and Fos or p53. These data suggest the possibility of down regulation of Fos expression by high levels of p53 protein. Further work is necessary to determine the mechanism of this interaction.
Aged ; Carcinoma, Basal Cell/chemistry/*genetics ; Comparative Study ; Female ; Gene Expression ; Genes, fos ; Genes, jun ; Genes, p53 ; Human ; Immunohistochemistry ; Male ; Middle Age ; Oncogene Protein p65(gag-jun)/analysis ; Oncogene Proteins v-fos/analysis ; *Oncogenes ; Protein p53/analysis ; Receptor, Epidermal Growth Factor/analysis ; Skin Neoplasms/chemistry/*genetics

BACKGROUND: The main polyphenol components in green tee are (-)-epicatechin, (-)-epigallocatechin, (-)-epicatechin gallate and (-)-epigallocatechin gallate (EGCG). It is well known that flavonoids such as catechins can be protective against inflammatory and cancer and cardiovascular diseases. These protective effects are largely due to their inhibition of some enzymes and antioxidative activities by scavenging free radicals. Ultraviolet(UV) exposure of the skin, particulary UVB (290-320nm), causes adverse biological effects, including alterations in cutaneous immune cells, photoaging and photocarcinogenesis. Several studies have shown that EGCG afforded protection against UVB-induced inflammatory responses and photocarcinogenesis in murine models. OBJECTIVE AND METHODS: In this study, we investigated the effects of EGCG on UVB irradiated human skin fibroblasts using viability test, thiobarbituric acid assay, propidium iodide(PI) stain, and western blot analyses and RT-PCR. RESULTS: Cell survival curves after UVB irradiation showed dose dependent decrement pattern by trypan blue exclusion assay. Only 42% of dermal fibroblasts survived at 150 mJ/cm2 UVB irradiation. The damage was associated with cell membrane lipid peroxidation, as shown by accumulation malondialdehyde(MDA). By pre-cultivation with EGCG (50nmol), a significant preventive effect was noted on the increase in the absolute number of surviving cells(up to 81.5% of cells survived) and the levels of MDA markedly decreased. Morphological changes associated with apoptotic cell death were easily distinguished by PI stain. Bases on our finding, we investgated the regulation of p53, p21, bax, bcl-2, cyclin D1, E, Cdk2, and PARP proteins by western blot analyses. The expression p53 protein was elevated by following UVB exposure which was inhibited by EGCG treatment. Using RT-PCR, the transcription of p53, fas and jun gene showed similar results which obtained by western blot analyses. CONCLUSION: EGCG, which have newly accepted as a potential UV protection properties, is effective membrane peroxidation inhibitor and prevent apoptotic changes when present in relevant concentration at the site of action beginning and during UVB irradiation. And the protective mechanism of EGCG against UVB-induced cell damage maybe, at least in part, related with p53, fas and jun pathway.
Blotting, Western ; Cardiovascular Diseases ; Catechin ; Cell Death ; Cell Membrane ; Cell Survival ; Cyclin D1 ; Fibroblasts* ; Flavonoids ; Free Radicals ; Genes, jun ; Humans* ; Lipid Peroxidation ; Membranes ; Propidium ; Skin* ; Trypan Blue

BACKGROUND: Human cytomegalovirus (HCMV) has the ability to activate the expression of many viral and cellular genes. The c-jun proto-oncogene has known to be induced at immediate early time of HCMV infection, however, the mechanism of up-regulation of the gene was not known. We found HCMV immediate-early (IE) 2 expression transactivate the c-jun promoter in human embryonal lung cell (HEL). METHODS: The c-jun promoter region between -117 and -59 contains binding sites for the transcription factors Sp1, CAAT, AP-1 like (ATF/CREB), and MEF2. We tried to map the sequences in the c-jun promoter responsible for activation of the promoter by HCMV IE2 expression. Transient expression assays were performed using various reporter plasmids containing the c-jun promoter-regulatory region linked to the luciferase gene and a plasmid expressing HCMV IE2 gene. RESULTS: Deletional and point mutational analysis showed that ATF, MEF2, and another down stream elements were involved in the up-regulation of c-jun promoter. Gel mobility shift assay showed that there are several factors in HEL cell nuclear extracts that specifically bind to these sites and in vitro translated IE2 could not move or supershift the specific bands. CONCLUSION: This study delineate the mechanism of c-jun up-regulation in HCMV infection and would give the clue for the possible contribution of HCMV in tumorigenesis.
Binding Sites ; Carcinogenesis ; Cytomegalovirus* ; Electrophoretic Mobility Shift Assay ; Genes, jun ; Humans* ; Luciferases ; Lung ; Plasmids ; Promoter Regions, Genetic ; Rivers ; Transcription Factor AP-1 ; Transcription Factors ; Up-Regulation

To elucidate the endocrine mechanism of human parturition, the expression of c-Jun and c-Fos mRNA were examined in relation to estrogen receptor (ER) and progesterone receptor (PR) in human myometrium. c-Jun mRNA was detected in all myometrial tissues (n=5) during labor but not before labor (n=5) and in oxytocin-resistant postterm pregnancy (n=3). c-Fos mRNA was detected in only one myometrial tissue from a woman in labor. The distribution and intensity of immunostaining for ER and PR were semiquantitatively scored. During the late pregnancies, no significant difference was seen in the receptor scores for myometrial ER and PR between the patients who experienced labor and those who did not. Receptor scores for ER and PR were significantly lower in postterm pregnancy than in late pregnancy, regardless of the labor status. These data suggest that there are no changes in ER and PR in human myometrium during parturition. On the other hand, postterm pregnancy is associated with low ER and PR. c-Jun, induced during labor without changes in ER and PR, may play a role as a signaling mechanism in human myometrium.
Adult ; Blotting, Northern ; Female ; Genes, jun/genetics* ; Human ; Immunohistochemistry ; Labor/metabolism* ; Myometrium/metabolism* ; Myometrium/cytology ; Pregnancy ; RNA, Messenger/analysis ; Receptors, Estrogen/metabolism* ; Receptors, Progesterone/metabolism* ; Reference Values

The Jun activation-domain binding protein 1 (Jab1) recognize a potential coactivator of activator protein 1 (AP-1) such as c-fos, c-jun transcription factor and the fifth subunit of the COP9 signalosome complex. Also, Jab1 activate the c-jun gene resulted cell proliferation. Not only a powerful tumor suppressor but also regulator of apoptosis negative cdk inhibitor p27(kip1) are involved in the cell cycle. This is Jab1 and p27(kip1) interact with each other, Jab1 accelerate p27(kip1) from nuclear to cytoplasm through ubiquitin/proteasome pathway. However, information about the relationship between Jab1 and p27(kip1) is not known much. Taken together, the results of this study identify function and structure of Jab1 and p27(kip1) were described in a recent article on the basis of relevant. Besides Jab1 and p27(kip1) will organize the relationship between the disease and women.
Apoptosis ; Breast Neoplasms ; Carrier Proteins* ; Cell Cycle ; Cell Proliferation ; Cyclin-Dependent Kinase Inhibitor p27 ; Cytoplasm ; Endometriosis ; Female ; Genes, jun ; Humans ; Ovarian Neoplasms ; Transcription Factor AP-1 ; Transcription Factors