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1.
Biofactors ; 45(2): 253-258, 2019 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-30537158

RESUMO

Human sirtuin 1 (hSIRT1) is a NAD+ -dependent deacetylase that regulates several cellular processes. Unlike resveratrol, natural polymeric phenolic compounds isolated from Vitaceae are mostly hSIRT1 inhibitors. The resveratrol tetramer, (+)-hopeaphenol ((+)-HP), and its geometric isomer, (-)-isohopeaphenol ((-)-iHP), were tested for inhibitory effects on purified hSIRT1 using a fluorescent derivative of peptide substrate p53-AMC (Fluor de Lys) and a cofactor NAD+ . The Lineweaver-Burk plots indicated that both (+)-HP and (-)-iHP were competitive inhibitors against NAD+ . Computer-assisted modeling of the binding of these molecules with hSIRT1 protein provided the most feasible conformation of the enzyme-inhibitor complex. © 2018 BioFactors, 45(2):253-258, 2019.


Assuntos
Polifenóis/farmacologia , Sirtuína 1/química , Sirtuína 1/metabolismo , Estilbenos/farmacologia , Humanos , Fenóis/química , Fenóis/farmacologia , Polifenóis/química , Ligação Proteica , Resveratrol/química , Resveratrol/farmacologia , Sirtuína 1/antagonistas & inibidores , Estilbenos/química
2.
FEBS Open Bio ; 8(3): 349-360, 2018 03.
Artigo em Inglês | MEDLINE | ID: mdl-29511612

RESUMO

Amyloid-ß (Aß), a primary component of amyloid plaques, has been widely associated with the pathogenesis of Alzheimer's disease. The Ca2+-binding protein regucalcin (RGN) plays multiple roles in maintaining cell functions by regulating intracellular calcium homeostasis, various signaling pathways, and gene expression systems. Here, we investigated the functional role of RGN against Aß-induced cytotoxicity in neuronally differentiated PC12 cells. Overexpression of RGN reduced Aß-induced apoptosis by reducing mitochondrial dysfunction and caspase activation. It also attenuated Aß-induced reactive oxygen species production and oxidative damage and decreased Aß-induced nitric oxide (NO) overproduction, upregulation of inducible NO synthase by nuclear factor-κB, and nitrosative damage. Interestingly, the genetic disruption of RGN increased the susceptibility of neuronally differentiated PC12 cells to Aß toxicity. Thus, RGN possesses antioxidant activity against Aß-induced oxidative and nitrosative stress and may play protective roles against Aß-induced neurotoxicity in Alzheimer's disease.

3.
J Nat Med ; 72(1): 260-266, 2018 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-29151157

RESUMO

Erypoegin K is an isoflavone isolated from the stem bark of Erythrina poeppigiana. It contains a furan group at the A-ring of the core isoflavone structure and can inhibit the activity of glyoxalase I, an enzyme that catalyzes the detoxification of methylglyoxal (MG), a by-product of glycolysis. In the present study, we found that erypoegin K has a potent cytotoxic effect on human leukemia HL-60 cells. Its cytotoxic effect was much stronger than that of a known glyoxalase I inhibitor S-p-bromobenzylglutathione cyclopentyl diester. Conversely, erypoegin K demonstrated weak cytotoxicity toward normal human peripheral lymphocytes. The treatment of HL-60 cells with erypoegin K significantly induced caspase-3 activity, whereas the pretreatment of the cells with caspase-3 inhibitor suppressed erypoegin K-induced cell death. Furthermore, nuclear condensation and apoptotic genome DNA fragmentation were observed in erypoegin K-treated HL-60 cells. These results indicated that the observed cell death was mediated by apoptosis. In addition, the toxic compound MG was highly accumulated in the culture medium of erypoegin K-treated HL-60 cells, suggesting that cell apoptosis was triggered by extracellular MG. The present study showed that erypoegin K has a potent apoptosis-inducing effect on cancerous cell lines, such as HL-60.


Assuntos
Benzofuranos/química , Erythrina/química , Células HL-60/química , Isoflavonas/química , Leucemia/tratamento farmacológico , Apoptose , Humanos , Leucemia/patologia
4.
Nat Prod Commun ; 10(9): 1581-4, 2015 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-26594764

RESUMO

It has been reported that many malignant human tissues, including breast, colon, and lung cancers, may show an elevated expression of glyoxalase I (GLO I). GLO I catalyzes the reaction to transform hemimercaptal, a compound formed from methylglyoxal (MG) and reduced glutathione, into S-D-lactoylglutathione, which is then converted to D-lactic acid by glyoxalase II. GLO I inhibitors are expected to be useful for inhibiting tumorigenesis through the accumulation of apoptosis-inducible MG in tumor cells. Here, we investigated the anti-proliferative activity of eight kinds of isoflavone isolated from Erythrina poeppigiana against the growth of HL-60 human leukemia cells from the viewpoint of GLO I inhibition. Of the compounds tested, the diprenyl isoflavone, isolupalbigenin, was shown to exhibit the highest anti-proliferative activity against HL-60 cells. Upon the treatment of HL-60 cells with isolupalbigenin, MG was significantly accumulated in the culture medium, and the caspase 3 activity of the cell lysate was elevated in a time-dependent manner. Thus, it is suggested that isolupalbigenin inhibits the enzyme GLO I, resulting in MG accumulation in the medium, and leading to cell apoptosis. Isolupalbigenin, with two prenyl groups in its A- and B-rings, might be expected to become a potent leading compound for the development of anticancer agents.


Assuntos
Antineoplásicos Fitogênicos/farmacologia , Erythrina/química , Isoflavonas/farmacologia , Lactoilglutationa Liase/antagonistas & inibidores , Lactoilglutationa Liase/metabolismo , Antineoplásicos Fitogênicos/química , Sobrevivência Celular , Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Regulação Enzimológica da Expressão Gênica , Células HL-60 , Humanos , Isoflavonas/química , Lactoilglutationa Liase/genética , Estrutura Molecular
5.
J Nat Med ; 68(3): 636-42, 2014 Jul.
Artigo em Inglês | MEDLINE | ID: mdl-24756815

RESUMO

A novel isoflavone, erythgianin A (1), along with nine known compounds 2-10, was isolated from the stem bark of Erythrina poeppigiana (Leguminosae). The unusual isoflavone structure of 1, possessing a highly oxidized 3″,4″-dihydroxy-2″-hydroxymethyl-2″-methyl-2″,3″-dihydropyrano substituent, was determined on the basis of spectroscopic analyses. All of the isolated compounds were evaluated for their in vitro inhibitory activity toward human glyoxalase I. Among the isolates, isolupalbigenin (10) with two prenyl groups showed the highest inhibitory activity.


Assuntos
Inibidores Enzimáticos/química , Inibidores Enzimáticos/farmacologia , Erythrina/química , Lactoilglutationa Liase/antagonistas & inibidores , Fenóis/química , Fenóis/farmacologia , Humanos , Isoflavonas/química , Isoflavonas/farmacologia , Casca de Planta/química
6.
J Pharm Pharmacol ; 65(8): 1204-13, 2013 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-23837588

RESUMO

OBJECTIVES: The aim of this study was to examine the mechanism underlying the inhibitory effect of our synthesized carbazolequinone derivatives on nitric oxide (NO) production in activated macrophages. METHODS: Lipopolysaccharide (LPS) and interferon-γ (IFN-γ)-stimulated RAW264.7 macrophages were treated with carbazolequinone derivatives. The NO and prostaglandin E2 (PGE2 ) levels in cell culture supernatants fractions were measured by Greiss and ELISA assay, respectively. The expression of inducible NO synthase (iNOS) and cyclooxygenase-2 (COX-2) was assessed by the real-time RT-PCR method. Nuclear factor kappa B (NF-κB) activation was detected by an NF-κB-dependent luciferase reporter assay. KEY FINDINGS: Our synthesized carbazolequinone derivatives (7-methoxy-2-methylcarbazole-1,4-quinone, 6-methoxy-2-methylcarbazole-1,4-quinone and 6-chloro-2-methylcarbazole-1,4-quinone) significantly inhibited LPS/IFN-γ-induced NO production and iNOS expression in RAW264.7 cells. They also inhibited the LPS/IFN-γ-mediated induction of COX-2 expression and PGE2 production. In addition, the LPS/IFN-γ-induced transcription activity of NF-κB was attenuated. Using the RAW264.7-tsAM5NE co-culture system, we found that these carbazolequinone derivatives protected neuronally differentiated tsAM5NE cells from NO-induced cell death by inhibiting the production of NO. CONCLUSIONS: These results suggest that the three carbazolequinone derivatives inhibit LPS/IFN-γ-induced NO production via iNOS and COX-2 downregulation due to NF-κB inhibition. Therefore, these three carbazolequinone derivatives may be useful for developing a new drug against NO-mediated neurodegenerative diseases.


Assuntos
Anti-Inflamatórios/farmacologia , Carbazóis/farmacologia , Interferon gama/farmacologia , Lipopolissacarídeos/farmacologia , Macrófagos/efeitos dos fármacos , Óxido Nítrico/antagonistas & inibidores , Quinonas/farmacologia , Animais , Anti-Inflamatórios/química , Anti-Inflamatórios/isolamento & purificação , Carbazóis/química , Carbazóis/isolamento & purificação , Técnicas de Cultura de Células , Linhagem Celular , Ciclo-Oxigenase 2/genética , Macrófagos/metabolismo , Camundongos , Estrutura Molecular , Murraya/química , Óxido Nítrico/biossíntese , Óxido Nítrico Sintase Tipo II/genética , Quinonas/química , Quinonas/isolamento & purificação
7.
J Nat Med ; 65(2): 353-9, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21246298

RESUMO

It is well known that inflammation is associated with various neurodegenerative diseases, such as Parkinson's disease and Alzheimer's disease. An inflammatory mediator, nitric oxide (NO), is produced by inducible NO synthase (iNOS) in microglia and seems to be one of the possible causes of neurodegeneration. Several natural and synthetic compounds which exert anti-inflammatory effects by inhibiting NO production have been reported to date. The aim of this work was to investigate whether any of the 6 terpenoid coumarins (methyl galbanate, galbanic acid, farnesiferol A, badrakemone, umbelliprenin, and aurapten) isolated from Ferula szowitsiana DC. have inhibitory activity against NO production in RAW264.7 mouse macrophage cells stimulated with lipopolysaccharide (LPS) and interferon-γ (IFN-γ). Of the 6 terpenoid coumarins tested, methyl galbanate significantly decreased NO production in LPS/IFN-γ-stimulated RAW264.7 cells. In the presence of methyl galbanate, LPS/IFN-γ-induced iNOS mRNA expression was significantly decreased to 52% of the level found with LPS/IFN-γ stimulation alone. Methyl galbanate slightly attenuated COX-2 mRNA expression. Using the RAW264.7-tsAM5NE co-culture system, we showed that methyl galbanate protected neuronally differentiated tsAM5NE cells from NO-induced cell death by inhibiting the production of NO. Our finding suggests that methyl galbanate may be useful for developing a new drug against neurodegenerative diseases.


Assuntos
Cumarínicos/química , Cumarínicos/farmacologia , Macrófagos/efeitos dos fármacos , Macrófagos/metabolismo , Óxido Nítrico/biossíntese , Terpenos/química , Terpenos/farmacologia , Animais , Western Blotting , Linhagem Celular , Interferon gama/farmacologia , Lipopolissacarídeos/farmacologia , Camundongos , Estrutura Molecular , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Sesquiterpenos/química , Sesquiterpenos/farmacologia , Umbeliferonas/química , Umbeliferonas/farmacologia
8.
Cell Biol Int ; 35(4): 325-34, 2011 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-21029049

RESUMO

We established a clonal adrenal medullary cell line, named tsAM5NE, from transgenic mice harbouring the temperature-sensitive Simian virus 40 large T-antigen gene, under the control of the tyrosine hydroxylase promoter. tsAM5NE cells conditionally grew at a permissive temperature of 33°C and exhibited the noradrenergic chromaffin cell phenotype. To understand the characteristics of tsAM5NE cells, we first examined the responsiveness of the cells to ligands of the GDNF (glial cell line-derived neurotrophic factor) family. tsAM5NE cells proliferated at the permissive temperature of 33°C in response to either GDNF or neurturin, but not artemin or persephin. At the non-permissive temperature of 39°C, GDNF or neurturin caused tsAM5NE cells to differentiate into neuron-like cells; however, the differentiated cells died in a time-dependent manner. Interestingly, LIF (leukaemia inhibitory factor) did not affect the GDNF-mediated cell proliferation at 33°C, but promoted the survival and differentiation of GDNF-treated cells at 39°C. In the presence of GDNF plus LIF, the morphological change induced by the temperature shift was associated with up-regulated expression of neuronal markers, indicating that the cells had indeed undergone neuronal differentiation. Thus, we demonstrated that tsAM5NE cells had the capacity to terminally differentiate into neuron-like cells in response to GDNF plus LIF when the oncogene was inactivated by the temperature shift. Thus, this cell line provides a useful model system for studying the mechanisms regulating neuronal differentiation.


Assuntos
Glândulas Suprarrenais/citologia , Antígenos Transformantes de Poliomavirus/genética , Linhagem Celular Tumoral/citologia , Células Cromafins/citologia , Animais , Linhagem Celular Tumoral/metabolismo , Proliferação de Células , Células Cultivadas , Células Cromafins/metabolismo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Camundongos , Camundongos Transgênicos , Fatores de Crescimento Neural/metabolismo , Neurogênese , Neurônios/citologia
9.
Neurosci Lett ; 438(1): 42-7, 2008 Jun 13.
Artigo em Inglês | MEDLINE | ID: mdl-18455310

RESUMO

We recently established adrenal medullary cell line tsAM5D, which was immortalized by use of a temperature-sensitive mutant of the oncogene simian virus 40 large T-antigen. In the present study, when co-treated with glial cell line-derived neurotrophic factor (GDNF) and ciliary neurotrophic factor (CNTF), tsAM5D cells proliferated at the permissive temperature (33 degrees C) for the T-antigen expression and differentiated into neuron-like cells at the nonpermissive temperature (39 degrees C). Interestingly, in GDNF/CNTF-treated cultures, the addition of pan-specific transforming growth factor (TGF)-beta-neutralizing antibody did not affect the cell proliferation at 33 degrees C, but significantly reduced the survival of neuronally differentiated cells at 39 degrees C. Using real-time RT-PCR for analysis of GDNF/CNTF-treated cells, we found that the expression of mRNAs for TGF-beta1, TGF-beta2, and TGF-beta3 was up-regulated by the temperature shift. These results suggest that autocrine TGF-beta signaling is necessary for the survival of GDNF/CNTF-differentiated tsAM5D cells upon the temperature shift.


Assuntos
Medula Suprarrenal/crescimento & desenvolvimento , Medula Suprarrenal/metabolismo , Comunicação Autócrina/fisiologia , Células Cromafins/metabolismo , Neurônios/metabolismo , Fator de Crescimento Transformador beta/genética , Medula Suprarrenal/citologia , Animais , Antígenos Transformantes de Poliomavirus/genética , Antígenos Transformantes de Poliomavirus/metabolismo , Comunicação Autócrina/efeitos dos fármacos , Diferenciação Celular/efeitos dos fármacos , Diferenciação Celular/fisiologia , Linhagem Celular Transformada , Células Cromafins/citologia , Células Cromafins/efeitos dos fármacos , Fator Neurotrófico Ciliar/metabolismo , Fator Neurotrófico Ciliar/farmacologia , Fator Neurotrófico Derivado de Linhagem de Célula Glial/metabolismo , Fator Neurotrófico Derivado de Linhagem de Célula Glial/farmacologia , Camundongos , Neurônios/citologia , Neurônios/efeitos dos fármacos , Isoformas de Proteínas/genética , RNA Mensageiro/metabolismo , Temperatura Ambiente , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta2/genética , Fator de Crescimento Transformador beta3/genética , Regulação para Cima/genética
10.
J Neurosci Res ; 86(8): 1694-710, 2008 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-18293415

RESUMO

To understand the characteristics of tsAM5D cells immortalized with the temperature-sensitive simian virus 40 large T-antigen, we first examined the responsiveness of the cells to ligands of the glial cell line-derived neurotrophic factor (GDNF) family. tsAM5D cells proliferated at the permissive temperature of 33 degrees C in response to either GDNF or neurturin, but not persephin or artemin. At the nonpermissive temperature of 39 degrees C, GDNF or neurturin caused tsAM5D cells to differentiate into neuron-like cells; however, the differentiated cells died in a time-dependent manner. Interestingly, ciliary neurotrophic factor (CNTF) did not affect the GDNF-mediated cell proliferation at 33 degrees C but promoted the survival and differentiation of GDNF-treated cells at 39 degrees C. In the presence of GDNF plus CNTF, the morphological change induced by the temperature shift was associated with up-regulated expression of various neuronal marker genes, indicating that the cells had undergone neuronal differentiation. In addition, tsAM5D cells caused to differentiate by GDNF plus CNTF at 39 degrees C became dependent solely on nerve growth factor (NGF) for their survival and neurite outgrowth. Moreover, upon treatment with GDNF plus CNTF, the dopaminergic phenotype was suppressed by the temperature shift. Thus, we demonstrated that tsAM5D cells had the capacity to differentiate terminally into neuron-like cells in response to GDNF plus CNTF when the oncogene was inactivated by the temperature shift. This cell line provides a useful model system for studying the role of a variety of signaling molecules for GDNF/CNTF-induced neuronal differentiation.


Assuntos
Antígenos Transformantes de Poliomavirus/fisiologia , Diferenciação Celular/fisiologia , Células Cromafins/citologia , Fator Neurotrófico Ciliar/fisiologia , Fator Neurotrófico Derivado de Linhagem de Célula Glial/fisiologia , Neurônios/citologia , Glândulas Suprarrenais/citologia , Glândulas Suprarrenais/fisiologia , Animais , Morte Celular/fisiologia , Linhagem Celular Transformada , Células Cultivadas , Células Cromafins/fisiologia , Humanos , Fator de Crescimento Neural/fisiologia , Neurônios/fisiologia , Ratos , Temperatura Ambiente , Fatores de Tempo
11.
J Biol Chem ; 281(32): 22503-16, 2006 Aug 11.
Artigo em Inglês | MEDLINE | ID: mdl-16772303

RESUMO

We previously established the murine adrenal chromaffin cell line tsAM5D, which was immortalized with the temperature-sensitive simian virus 40 large T-antigen. tsAM5D cells have the capacity to differentiate into neuron-like cells in response to neurotrophic factors when the culture temperature is shifted from 33 to 39 degrees C. In this model system, the temperature shift in the absence of neurotrophic factors led to cell death. Hoechst staining analysis revealed that typical apoptotic nuclei appeared in a time-dependent manner after the temperature shift. Upon shifting to 39 degrees C, the degradation of T-antigen was accompanied by the transcriptional activation of p53 protein. Among the p53 target genes, death receptor 5 (DR5), which is the receptor for tumor necrosis factor-related apoptosis-inducing ligand (TRAIL), showed the highest level of induction. Interestingly, TRAIL-neutralizing antibody protected tsAM5D cells from the temperature shift-induced apoptotic cell death by blocking the activation of caspase-8 and -3, indicating the involvement of TRAIL-mediated death signaling in the temperature shift-induced apoptosis. Glial cell line-derived neurotrophic factor (GDNF) inhibited the TRAIL-mediated activation of caspase-8 in tsAM5D cells exposed to 39 degrees C and cooperated with basic fibroblast growth factor and ciliary neurotrophic factor. Interestingly, the temperature shift induced oligomerization of DR5, which is the earliest process necessary for transduction of the death signal. This oligomerization was inhibited by treatment with GDNF plus ciliary neurotrophic factor but not by that with GDNF alone or GDNF plus basic fibroblast growth factor. These results are discussed with respect to the intracellular mechanism underlying the protective function of neurotrophic factors against TRAIL-mediated death signaling.


Assuntos
Glândulas Suprarrenais/citologia , Proteínas Reguladoras de Apoptose/metabolismo , Apoptose , Células Cromafins/metabolismo , Glicoproteínas de Membrana/metabolismo , Fatores de Crescimento Neural/metabolismo , Fator de Necrose Tumoral alfa/metabolismo , Animais , Caspase 3 , Caspase 8 , Caspases/metabolismo , Diferenciação Celular , Inibidores Enzimáticos/farmacologia , Camundongos , Receptores do Ligante Indutor de Apoptose Relacionado a TNF , Receptores do Fator de Necrose Tumoral/metabolismo , Transdução de Sinais , Ligante Indutor de Apoptose Relacionado a TNF , Proteína Supressora de Tumor p53/metabolismo
12.
Exp Cell Res ; 303(2): 287-99, 2005 Feb 15.
Artigo em Inglês | MEDLINE | ID: mdl-15652343

RESUMO

TIS11, a member of the CCCH zinc finger protein family, was found to be distributed throughout cells with a preferential cytoplasmic localization when transiently expressed in COS-7 cells. Upon treatment with heat shock, TIS11 became localized in discrete particles in the cytoplasm of the transfectants. We showed the TIS11-positive particles to be stress granules (SGs), which are known to be formed in the cytoplasm of eukaryotic cells in response to environmental stresses. By deletion studies using the green fluorescent protein fusion system, we mapped a functional stress granule (SG) localization signal to a region containing two tandem repeats of the zinc finger motif of TIS11. Site-directed mutations of Tyr105/Tyr113, Gly109/Gly 114, and Phe119 in the first zinc finger motif diminished the ability of this TIS11 domain to direct SG localization. Importantly, when the zinc-chelating Cys residues in either the first or second zinc finger were mutated to Ala residues, the recruitment of the TIS11 zinc finger region to SG was significantly inhibited by the mutation and was completely abolished by the mutation in both zinc fingers. These results suggest that recruitment of TIS11 to heat shock-induced SG is governed by the tandem zinc finger domains of this protein.


Assuntos
Proteínas de Ligação a DNA/química , Proteínas de Ligação a DNA/metabolismo , Proteínas Imediatamente Precoces/química , Proteínas Imediatamente Precoces/metabolismo , RNA Mensageiro/metabolismo , Sequência de Aminoácidos , Animais , Sequência de Bases , Transporte Biológico Ativo , Células COS , Grânulos Citoplasmáticos/metabolismo , DNA Complementar/genética , Proteínas de Ligação a DNA/genética , Proteínas de Fluorescência Verde/química , Proteínas de Fluorescência Verde/genética , Proteínas de Fluorescência Verde/metabolismo , Resposta ao Choque Térmico , Proteínas Imediatamente Precoces/genética , Modelos Moleculares , Dados de Sequência Molecular , Mutagênese Sítio-Dirigida , Estabilidade de RNA , Ratos , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Transfecção , Tristetraprolina , Dedos de Zinco/genética
13.
J Neurochem ; 85(5): 1126-38, 2003 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-12753072

RESUMO

We established adrenal medullary cell lines from transgenic mice expressing an oncogene, the temperature-sensitive simian virus 40 large T-antigen, under the control of the tyrosine hydroxylase promoter. A clonal cell line, named tsAM5D, conditionally grew at a permissive temperature of 33 degrees C and exhibited the dopaminergic chromaffin cell phenotype as exemplified by the expression pattern of mRNA for catecholamine-synthesizing enzymes and secretory vesicle-associated proteins. tsAM5D cells proliferated at the permissive temperature in response to basic fibroblast growth factor (bFGF) and ciliary neurotrophic factor (CNTF). At a non-permissive temperature of 39 degrees C, bFGF and CNTF acted synergistically to differentiate tsAM5D cells into neuron-like cells. In addition, tsAM5D cells caused to differentiate by bFGF plus CNTF at 39 degrees C became dependent solely on nerve growth factor for their survival and showed markedly enhanced neurite outgrowth. In the presence of bFGF and CNTF, the morphological change induced by the temperature shift was associated with up-regulated expression of neuronal marker genes including neuron-specific enolase, growth-associated protein-43, microtubule-associated protein 2, neurofilament, and p75 neurotrophin receptor, indicating that the cells underwent neuronal differentiation. Thus, we demonstrated that tsAM5D cells could proliferate at permissive 33 degrees C, and also had the capacity to terminally differentiate into neuron-like cells in response to bFGF and CNTF when the oncogene was inactivated by shifting the temperature to non-permissive 39 degrees C. These results suggest that tsAM5D cells should be a good tool to allow a detailed study of mechanisms regulating neuronal differentiation.


Assuntos
Neoplasias das Glândulas Suprarrenais/genética , Glândulas Suprarrenais/citologia , Antígenos Transformantes de Poliomavirus/genética , Diferenciação Celular/efeitos dos fármacos , Células Cromafins/efeitos dos fármacos , Fatores de Crescimento Neural/farmacologia , Neoplasias das Glândulas Suprarrenais/patologia , Animais , Biomarcadores/análise , Divisão Celular/efeitos dos fármacos , Divisão Celular/genética , Células Cultivadas , Células Cromafins/citologia , Células Cromafins/fisiologia , Fator Neurotrófico Ciliar/farmacologia , Células Clonais , Fator 2 de Crescimento de Fibroblastos/farmacologia , Expressão Gênica/efeitos dos fármacos , Humanos , Camundongos , Camundongos Transgênicos , Regiões Promotoras Genéticas , Transdução de Sinais/fisiologia , Temperatura Ambiente , Tirosina 3-Mono-Oxigenase/genética
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