1.Regulation of c-fos gene expression in hepatic stellate cells by transforming growth factor beta.
Ai-Min WANG ; Chun-Gai REN ; Zhi-Jie MA ; Ying XIANG ; Zhi-Wei ZHANG ; Zhi-Jian ZHU ; Bao-En WANG
Journal of Southern Medical University 2009;29(7):1408-1409
OBJECTIVETo investigate the effects of transforming growth factor beta (TGF ) on c-fos gene expression in hepatic stellate cells.
METHODSHepatic stellate cells (HSC-T6) were cultured in the medium containing different concentrations of TGF (0.2, 1, and 5 ng/ml), and cells were collected at different time points of incubation (8, 24, 48, and 72 h). The total RNA of the HSCs was isolated and c-fos gene expression level were measured by reverse transcription polymerase chain reaction.
RESULTSc-fos gene expression levels of HSCs cultured in the presence of low (0.2 ng/ml), moderate (1 ng/ml) and high (5 ng/ml) concentrations of TGF for 8, 24, 48 and 72 h were significantly greater than those of control group. The c-fos gene expression levels of HSCs increased gradually with the increment of TGF concentration, and significant differences in c-fos gene expression were found between the 3TGF groups.
CONCLUSIONTGF strongly up-regulates c-fos gene expression in hepatic stellate cells.
Animals ; Cells, Cultured ; Gene Expression ; drug effects ; Genes, fos ; Hepatic Stellate Cells ; drug effects ; Proto-Oncogene Proteins c-fos ; genetics ; metabolism ; Rats ; Transforming Growth Factor beta ; pharmacology
2.Effects of cadmium on hepatocellular DNA damage, proto-oncogene expression and apoptosis in rats.
Ri-An YU ; Ling-Fei HE ; Xue-Min CHEN
Biomedical and Environmental Sciences 2007;20(2):146-153
OBJECTIVETo study the effects of cadmium on hepatocellular DNA damage, expression of proto-oncogenes c-myc, c-fos, and c-jun as well as apoptosis in rats.
METHODSCadmium chloride at the doses of 5, 10, and 20 micromol/kg was given to rats by i.p. and there were 5 male SD rats in each group. Hepatocellular DNA damage was measured by single cell gel electrophoresis (or comet assay), while expression of proto-oncogenes c-myc, c-fos, and c-jun in rat hepatocytes were measured by Northern dot hybridization. C-Myc, c-Fos, and c-Jun were detected with immuno-histochemical method. Hepatocellular apoptosis was determined by TUNEL (TdT-mediated dUTP Nick End Labelling) and flow cytometry.
RESULTSAt the doses of 5, 10, and 20 micromol/kg, cadmium chloride induced DNA damage in rat hepatocytes and the rates of comet cells were 50.20%, 88.40%, and 93.80%, respectively. Results also showed an obvious dose-response relationship between the rates of comet cells and the dose of cadmium chloride (r = 0.9172, P < 0.01). Cadmium chloride at the doses of 5, 10, and 20 micromol/kg induced expression of proto-oncogenes c-myc, c-fos, and c-jun. The positive brown-yellow signal for c-myc, c-fos, and c-jun was mainly located in the cytoplasm of hepatocytes with immunohistochemical method. TUNEL-positive cells were detected in cadmium-treated rat livers. Apoptotic rates (%) of cadmium-treated liver cells at the doses of 5, 10, and 20 micromol/kg were (17.24 +/- 2.98), (20.58 +/- 1.35), and (24.06 +/- 1.77) respectively, being significantly higher than those in the control. The results also displayed an obvious dose-response relationship between apoptotic rates and the dose of cadmium chloride (r = 0.8619, P < 0.05).
CONCLUSIONCadmium at 5-20 micromol/kg can induce hepatocellular DNA damage, expression of proto-oncogenes c-myc, c-fos, and c-jun as well as apoptosis in rats.
Animals ; Apoptosis ; drug effects ; Cadmium ; toxicity ; DNA Damage ; Gene Expression Regulation ; drug effects ; Hepatocytes ; cytology ; drug effects ; metabolism ; Male ; Proto-Oncogene Proteins ; genetics ; metabolism ; Proto-Oncogene Proteins c-fos ; genetics ; metabolism ; Proto-Oncogene Proteins c-jun ; genetics ; metabolism ; Proto-Oncogene Proteins c-myc ; genetics ; metabolism ; Rats ; Rats, Sprague-Dawley
3.Preparation and application of goat deltafosB gene expression product antibody.
Huiling ZHENG ; Zhenzhen ZHU ; Junhui AN ; Zhenyu YANG ; Ruifang XING ; Linhui YAN
Chinese Journal of Biotechnology 2010;26(12):1704-1709
deltaFosB, a naturally occurring truncated isform of fosB gene, existed in many tissues stably and played an important role in formation and differentiation of adipocyte and osteoblast. deltaFosB may be related to the metabolism of calcium in bone and mammary gland and regulate the signal pathway of calcium transfer from bone to mammary gland. We first sub-cloned deltafosB gene of goat into the vector pET32a to construct prokaryotic expression vector pET32a-deltafosB. Then we induced for deltafosB gene expression efficiently by IPTG. Finally we immunized the adult rabbits with purified recombinant deltaFosB to prepare rabbit anti-goat deltaFosB polyclonal antibody. iELISA analysis showed the antibody with the titer of 1:51 200, and Western blotting result showed that the antibody could specifically detect the deltaFosB protein expressed in prokaryotic cell and HEK-293 cell, respectively. Further Western blotting assay showed that deltaFosB expressed in various tissues of goat in vivo.
Animals
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Antibodies, Monoclonal
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biosynthesis
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Cloning, Molecular
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Escherichia coli
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genetics
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metabolism
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Gene Expression
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Goats
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Proto-Oncogene Proteins c-fos
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biosynthesis
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genetics
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immunology
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Rabbits
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Recombinant Proteins
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biosynthesis
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genetics
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immunology
4.Expression of c-fos, OPG, OPGL in rabbit mandibular distraction osteogenesis zone.
Wei-li GE ; Zhi-jian XIE ; Jian-feng HE
Journal of Zhejiang University. Medical sciences 2006;35(5):496-500
OBJECTIVETo evaluate the possible signal transduction mechanism of the mechanical stress induced by the distraction procedure in osteocytes.
METHODSAn animal model of mandibular distraction osteogenesis in rabbits was established. The expressions of c-fos, OPG and OPGL were detected by ultrasensitive S-P immunohistochemical method.
RESULTAt 4 and 8 days after distraction, distraction zone showed strong positive staining of c-fos, which were apparently higher than that in distraction zone of 2, 4 and 6 weeks after consolidation. At 4 and 8 days after distraction and 2 weeks after consolidation, the expression of OPG was strong, and then wore off gradually at 4 and 6 weeks after consolidation. Weak signals of OPGL could be detected at 6 weeks after consolidation only.
CONCLUSIONc-fos, OPG and OPGL are important regulators in distraction osteogenesis. c-fos is interrelated with the mechanical stress induced by the distraction procedure closely, OPG promotes new bone formation, while OPGL plays a more active role in bone remodeling.
Animals ; Mandible ; cytology ; metabolism ; Osteocytes ; metabolism ; Osteogenesis, Distraction ; Osteoprotegerin ; biosynthesis ; genetics ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; RANK Ligand ; biosynthesis ; genetics ; Rabbits ; Random Allocation
5.Expressions of c-Fos and NADPH-d in the related brainstem during vestibular compensation.
Acta Academiae Medicinae Sinicae 2008;30(6):655-658
OBJECTIVETo study the mechanism of vestibular compensation and to observe the changes of c-Fos and NADPH-d expressions in the brainstem of the vestibular deafferentation rats in static status or following angular acceleration stimulation.
METHODSTotally 60 SD rats were randomly divided into control group (labyrinthine intact), complete unilateral vestibular deafferentation (UVD) group, simultaneous complete bilateral vestibular deafferentation (BVD) group (n = 20 in each group). Subgroups (n = 10 in each subgroup) were set for static status or following angular acceleration stimulation in each group. Double labeling with histochemistry-immunohistochemistry was performed to observe c-Fos/NADPH-d neurons.
RESULTSNo positive c-Fos/NADPH-d expression was observed in the both sides of medial vestibular nucleus (MVN) and prepositus hypoglossi (PrH) of normal rats in static status and BVD rats whether following canal rotation or not. c-Fos/ NADPH-d expression was observed in the ipsilesional MVN and the contralesional PrH of UVD rats. However, c-Fos/NADPH-d were detected in both sides of MVN and PrH in UVD rats and normal rats following angular acceleration stimulation.
CONCLUSIONIn the ipsilesional MVN and the contralesional PrH, c-Fos plays an important role in vestibular compensation, in which nitric oxide acts as a key neurotransmitter.
Animals ; Brain Stem ; metabolism ; Dihydrolipoamide Dehydrogenase ; genetics ; metabolism ; Female ; Gene Expression ; Male ; Proto-Oncogene Proteins c-fos ; genetics ; metabolism ; Random Allocation ; Rats ; Rats, Sprague-Dawley ; Vestibule, Labyrinth ; physiology
7.Effect of heparin-derived oligosaccharide on vascular smooth muscle cell proliferation through inhibition of PKC-alpha expression.
Li LI ; Ting GAO ; Shu-ying HE ; Guang-lin XU ; Li-na YANG
Acta Pharmaceutica Sinica 2012;47(8):993-1000
In this study, the effect of heparin-derived oligosaccharide (HDO) on bovine vascular smooth muscle cell (VSMC) proliferation and signal transduction mechanism involved were investigated. The levels of PKC-alpha protein and mRNA were determined by cell-based ELISA, RT-PCR, Western blotting and immunocytochemical methods. Meanwhile, mRNA levels of c-jun, c-myc and c-fos were assayed by RT-PCR method. The results showed that HDO inhibited newborn calf serum (NCS)-induced expression of PKC-alpha and proto-oncogenes, which may be one of the mechanisms for the inhibition of VSMC proliferation by HDO. Flow cytometry analysis indicated that HDO blocked NCS-induced cell cycle progression by arresting cells at G0/G1 phase. The results imply that HDO inhibits VSMC proliferation by moderating the gene level of PKC-alpha, eventually inhibiting proto-oncogene mRNA expression and blocking G1/S transition.
Animals
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Cattle
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Cell Cycle
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drug effects
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Cell Proliferation
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drug effects
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Cells, Cultured
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G1 Phase
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drug effects
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Heparin
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pharmacology
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Muscle, Smooth, Vascular
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cytology
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metabolism
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Oligosaccharides
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pharmacology
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Protein Kinase C-alpha
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genetics
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metabolism
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Proto-Oncogene Proteins c-fos
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genetics
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metabolism
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Proto-Oncogene Proteins c-jun
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genetics
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metabolism
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Proto-Oncogene Proteins c-myc
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genetics
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metabolism
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RNA, Messenger
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metabolism
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Signal Transduction
8.Growth factors may enhance c-fos and c-jun gene expressions of hepatic stellate cells.
Ai-Min WANG ; Chun-Hai REN ; Ying XIANG
Chinese Journal of Hepatology 2008;16(12):902-904
OBJECTIVETo investigate the effects of growth factors on c-fos and c-jun gene expressions in hepatic stellate cells.
METHODSHepatic stellate cell (HSC) T6 was cultured in media containing different concentrations of platelet derived growth factor (PDGF) (8 ng/ml, 40 ng/ml or 200 ng/ml) and transforming growth factor (TGF) beta (0.2 ng/ml, 1 ng/ml or 5 ng/ml) and the cells were collected at different incubation periods (8, 24, 48 or 72 h). Total RNA of the HSC was isolated and c-fos and c-jun gene expression levels were measured by reverse transcription polymerase chain reaction.
RESULTSC-fos gene expression levels of the HSC cultured in low (8 ng/ml), medium (40 ng/ml) and high (200 ng/ml) concentrations of PDGF were all much higher than those of the control group after exposure to PDGF at 8, 24, 48 or 72 h. The c-fos gene expression levels of the HSC increased as the dosage of PDGF increased and there were significant differences of c-fos gene expression among the three PDGF groups. C-jun gene expression levels of the HSC in low (0.2 ng/ml), medium (1.0 ng/ml) and high (5.0 ng/ml) concentrations of TGF beta groups were much higher than those of the control group after exposure to TGF beta at 8, 24, 48 or 72 h. The c-jun gene expression levels of the HSC increased as the dosage of TGF beta increased and there were significant differences of c-jun gene expression among the three TGF beta groups.
CONCLUSIONPDGF and TGF beta can strongly up-regulate c-fos and c-jun gene expressions in hepatic stellate T6 cells.
Cells, Cultured ; Gene Expression ; Hepatic Stellate Cells ; drug effects ; metabolism ; Humans ; Platelet-Derived Growth Factor ; pharmacology ; Proto-Oncogene Proteins c-fos ; genetics ; Proto-Oncogene Proteins c-jun ; genetics ; Transforming Growth Factor beta ; pharmacology
10.Effect of mailuoning on C-Fos protein expression in rats with cerebral infarction.
Zhi-li GUO ; Ping LIU ; Yong-hong CHEN ; Jian SHANG ; Liubao ZHANG ; Chunning SHENG ; Yun LUO
Chinese Journal of Integrated Traditional and Western Medicine 2004;24(9):831-834
OBJECTIVETo investigate the protective effect of mailuoning (MLN) on nerve cells after cerebral infarction induced by photochemistry.
METHODSEighty SD rats were divided into three groups, the control group (n = 20), the model group (n = 30) and the MLN group (n = 30). Focal cerebro-ischemia induced by photothrombosis in adult rat was used as a model. Changes of C-Fos protein expression before and after MLN treatment were observed using immunohistochemistry, computerized imaging technique and transmission electron microscopy (TEM).
RESULTSC-Fos positive cells located in the transitional zone between the necrotic core and normal cortex. C-Fos protein expression began to show 3 hrs after cerebral infarction, peaked at 6 hrs. As compared with the model group, C-Fos expression was significantly reduced in the MLN group (P<0.01).
CONCLUSIONMLN could markedly reduce the injury of nerve cell in the transitional zone, the protection may be related with its inhibition on C-Fos protein expression.
Animals ; Cerebral Infarction ; etiology ; genetics ; metabolism ; Drugs, Chinese Herbal ; pharmacology ; Male ; Neuroprotective Agents ; pharmacology ; Photochemistry ; Proto-Oncogene Proteins c-fos ; biosynthesis ; genetics ; Rats ; Rats, Sprague-Dawley