Your browser doesn't support javascript.
loading
Mostrar: 20 | 50 | 100
Resultados 1 - 20 de 36
Filtrar
1.
J Investig Allergol Clin Immunol ; 33(4): 263-270, 2023 Jul 27.
Artigo em Inglês | MEDLINE | ID: mdl-35166674

RESUMO

BACKGROUND AND OBJECTIVE: Cartilage-hair hypoplasia (CHH) syndrome is a rare autosomal recessive syndrome associated with skeletal dysplasia, varying degrees of combined immunodeficiency (CID), short stature, hair hypoplasia, macrocytic anemia, increased risk of malignancies, and Hirschsprung disease. To provide clinical and immunological insights obtained from 2 unrelated patients who displayed clinical characteristics of CHH. METHODS: Two patients with suspected CHH syndrome due to skeletal dysplasia and immunodeficiency underwent an immunological and genetic work-up using flow cytometry, next-generation sequencing (NGS) of the immune repertoire, and Sanger sequencing to identify the underlying defects. RESULTS: Patient 1 presented with low birth weight and skeletal dysplasia. Newborn screening was suggestive of T-cell immunodeficiency, as T-cell receptor excision circle levels were undetectable. Both the T-cell receptor (TCR) Vß and TCR-g (TRG) repertoires were restricted, with evidence of clonal expansion. Genetic analysis identified compound heterozygous RMRP variants inherited from both parents. Patient 2 presented with recurrent lung and gastrointestinal infections, skeletal dysplasia, failure to thrive, and hepatomegaly. The polyclonal pattern of the TCRß repertoire was normal, with only slight overexpression of TCR-ßV20 and restricted expression of Vßs. TRG expressed a normal diverse repertoire, similar to that of the healthy control sample. Genetic analysis identified biallelic novel regulatory variants in RMRP. Both parents are carriers of this mutation. CONCLUSION: Our findings demonstrate how the immunological work-up, supported by genetic findings, can dramatically change treatment and future outcome in patients with the same clinical syndrome.


Assuntos
Doença de Hirschsprung , Síndromes de Imunodeficiência , Recém-Nascido , Humanos , Doença de Hirschsprung/genética , Doença de Hirschsprung/complicações , Doença de Hirschsprung/patologia , Síndromes de Imunodeficiência/genética , Cabelo/anormalidades , Cabelo/patologia , Receptores de Antígenos de Linfócitos T/genética , Progressão da Doença
2.
Clin Oncol (R Coll Radiol) ; 32(9): 579-590, 2020 09.
Artigo em Inglês | MEDLINE | ID: mdl-32299722

RESUMO

Diabetes and cancer are common conditions highly prevalent in the general population. The co-existence of diabetes and cancer in a patient is therefore not unexpected. Diabetes increases the risk of mortality from cancer and morbidity from the treatment of cancer. Furthermore, many cancer chemotherapeutic regimens increase glucose levels, especially those involving glucocorticoids. Many clinical oncologists will deal with patients with diabetes in their clinical work, and some working knowledge of diabetes diagnosis and management is helpful when managing such patients. This overview aims to summarise the clinical diagnosis and management of diabetes, review the potential links between diabetes and cancer, and provide some practical guidance on the management of hyperglycaemia in patients undergoing cancer therapy.


Assuntos
Antineoplásicos/efeitos adversos , Complicações do Diabetes/diagnóstico , Complicações do Diabetes/tratamento farmacológico , Diabetes Mellitus/diagnóstico , Diabetes Mellitus/tratamento farmacológico , Neoplasias/tratamento farmacológico , Oncologistas/normas , Guias de Prática Clínica como Assunto/normas , Complicações do Diabetes/induzido quimicamente , Diabetes Mellitus/induzido quimicamente , Humanos
4.
Cytopathology ; 24(2): 99-104, 2013 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-22612864

RESUMO

OBJECTIVE: ThinPrep (TP), one of the Food and Drug Administration-approved liquid-based cytology (LBC) preparations, is widely used for gynaecological and non-gynaecological cytology samples. A unique physical artefact caused by the compression at the periphery in TP slides has not been adequately evaluated to date. METHODS: We processed four established tumour cell lines (MKN28, MKN45, KG-1 and NB4) and mononuclear cells isolated from whole blood over Ficoll-Plaque for TP preparations. For this part of the study, we included five normal cervical LBC preparations. We then auto-counted and auto-measured the area, mean grey value and Feret's diameter in both the inner disc and peripheral rim of the preparations by image morphometry. In addition, we compared the distribution of atypical cell groups in the peripheral rim and inner disc of 132 lung aspirates, 80 thyroid aspirates, 212 cerebrospinal fluids (CSFs) and 50 gynaecological samples. RESULTS: The areas and Feret's diameters of the cytoplasm in the peripheral compressed rim area were statistically larger than those of cells in the inner disc. The mean grey values of cells (cytoplasm and nucleus) in the peripheral compression rim were also smaller than those in the inner disc cells, leading to decreases in nuclear and cytoplasmic chromatism. Except for the mean grey values, the differences were not significant in the cervical samples. CONCLUSIONS: Cellular morphology may be markedly distorted in the peripheral rim, regardless of cell malignancy, which may lead to the misinterpretation of cells during the screening. Accordingly, cytological diagnosis based on the findings within the peripheral rim should take this phenomenon into account. Compressed cells found in the peripheral rim should be interpreted with caution when TP slides are used for cytopathological diagnosis.


Assuntos
Biópsia por Agulha Fina/métodos , Citodiagnóstico/métodos , Processamento de Imagem Assistida por Computador/métodos , Linhagem Celular Tumoral , Colo do Útero/patologia , Feminino , Humanos , Pulmão/patologia , Masculino , Gravidez , Glândula Tireoide/patologia , Esfregaço Vaginal
5.
Endoscopy ; 44(9): 819-24, 2012 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-22791587

RESUMO

BACKGROUND AND STUDY AIMS: The incidence of residual stones after mechanical lithotripsy for retained common bile duct (CBD) stones is relatively high. Peroral cholangioscopy using a mother-baby system may be useful for confirming complete extraction of stones, but has several limitations regarding routine use. We evaluated the role of direct peroral cholangioscopy (DPOC) using an ultraslim upper endoscope for the evaluation and removal of residual CBD stones after mechanical lithotripsy. PATIENTS AND METHODS: From August 2006 to November 2010, 48 patients who had undergone mechanical lithotripsy for retained CBD stones with no evidence of filling defects in balloon cholangiography were recruited. The bile duct was inspected by DPOC after balloon cholangiography. Detected residual CBD stones were directly retrieved with a basket or balloon catheter under DPOC. The incidence of residual stones detected by DPOC, and the success rate of residual stone retrieval under DPOC were investigated. RESULTS: DPOC was successfully performed in 46 of the 48 patients (95.8%). Of these, 13 patients (28.3%) had residual CBD stones (mean number 1.4, range 1-3; mean diameter 4.5 mm, range 2.3-9.6). The residual stones were removed directly under DPOC in 11 of these patients (84.6%). There were no complications associated with DPOC or stone removal. CONCLUSION: DPOC using an ultraslim upper endoscope is a useful endoscopic procedure for the evaluation and extraction of residual stones after mechanical lithotripsy for retained CBD stones.


Assuntos
Endoscopia do Sistema Digestório/métodos , Cálculos Biliares/terapia , Adulto , Idoso , Idoso de 80 Anos ou mais , Colangiopancreatografia Retrógrada Endoscópica , Endoscopia do Sistema Digestório/instrumentação , Feminino , Seguimentos , Cálculos Biliares/diagnóstico , Humanos , Litotripsia , Masculino , Pessoa de Meia-Idade
6.
Poult Sci ; 91(1): 66-73, 2012 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-22184430

RESUMO

Polyphenolic compounds present in green tea, particularly catechins, are known to have strong anti-influenza activity. The goal of this study was to determine whether green tea by-products could function as an alternative to common antivirals in animals compared to original green tea. Inhibition of viral cytopathic effects ascertained by neutral red dye uptake was examined with 50% effective (virus-inhibitory) concentrations (EC50)determined. Against the H1N1 virus A/NWS/33, we found the anti-influenza activity of green tea by-products (EC50 = 6.36 µg/mL) to be equivalent to that of original green tea (EC50= 6.72 µg/mL). The anti-influenza activity of green tea by-products was further examined in mouse and chicken influenza infection models. In mice, oral administration of green tea by-products reduced viral titers in the lungs in the early phase of infection, but they could not protect these animals from disease and death. In contrast, therapeutic administration of green tea by-products via feed or water supplement resulted in a dose-dependent significant antiviral effect in chickens, with a dose of 10 g/kg of feed being the most effective (P < 0.001). We also demonstrated that unidentified hexane-soluble fractions of green tea by-products possessed strong anti-influenza activity, in addition to ethyl acetate-soluble fractions, including catechins. This study revealed green tea by-product extracts to be a promising novel antiviral resource for animals.


Assuntos
Antivirais/administração & dosagem , Camellia sinensis/química , Vírus da Influenza A Subtipo H1N1/efeitos dos fármacos , Vírus da Influenza A Subtipo H9N2/efeitos dos fármacos , Infecções por Orthomyxoviridae/veterinária , Extratos Vegetais/administração & dosagem , Administração Intranasal/veterinária , Administração Oral , Animais , Antivirais/química , Antivirais/farmacologia , Catequina/análogos & derivados , Catequina/química , Catequina/farmacologia , Linhagem Celular , Galinhas , Testes de Inibição da Hemaglutinação/veterinária , Camundongos , Camundongos Endogâmicos BALB C , Neuraminidase/antagonistas & inibidores , Vermelho Neutro/química , Infecções por Orthomyxoviridae/tratamento farmacológico , Extratos Vegetais/química , Extratos Vegetais/farmacologia , Folhas de Planta/química , Reação em Cadeia da Polimerase Via Transcriptase Reversa/veterinária , Organismos Livres de Patógenos Específicos
7.
Poult Sci ; 90(5): 1020-2, 2011 May.
Artigo em Inglês | MEDLINE | ID: mdl-21489949

RESUMO

The frequent economic losses incurred with H9N2 low pathogenic avian influenza viruses (LPAI) infection have raised serious concerns for the poultry industry. A 1-dose regimen with inactivated H9N2 LPAI vaccine could not prevent vaccinated poultry from becoming infected and from shedding wild viruses. A study was conducted to determine whether a 2-dose regimen of inactivated H9N2 LPAI vaccine could enhance the immunologic response in chickens. Such gel-primed and mineral oil-boosted regimen has produced encouraging results associated with improved immune responses to an H9N2 LPAI. This strategy could be cost effective and helpful for preventing avian influenza virus in the poultry industry.


Assuntos
Galinhas , Vírus da Influenza A Subtipo H9N2/imunologia , Vacinas contra Influenza/imunologia , Influenza Aviária/prevenção & controle , Adjuvantes Imunológicos , Animais , Anticorpos Antivirais/sangue , Géis , Esquemas de Imunização , Imunização Secundária , Óleo Mineral , Organismos Livres de Patógenos Específicos , Vacinas de Produtos Inativados
9.
Mol Cell Biol ; 21(22): 7696-706, 2001 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-11604505

RESUMO

Determination and differentiation of skeletal muscle precursors requires cell-cell contact, but the full range of cell surface proteins that mediate this requirement and the mechanisms by which they work are not known. To identify participants in cell contact-mediated regulation of myogenesis, genes that encode secreted proteins specifically upregulated during differentiation of C2C12 myoblasts were identified by the yeast signal sequence trap method (K. A. Jacobs, L. A. Collins-Racie, M. Colbert, M. Duckett, M. Golden-Fleet, K. Kelleher, R. Kriz, E. R. La Vallie, D. Merberg, V. Spaulding, J. Stover, M. J. Williamson, and J. M. McCoy, Gene 198:289-296, 1997), followed by RNA expression analysis. We report here the identification of CD164 as a gene expressed in proliferating C2C12 cells that is upregulated during differentiation. CD164 encodes a widely expressed cell surface sialomucin that has been implicated in regulation of cell proliferation and adhesion during hematopoiesis. Stable overexpression of CD164 in C2C12 and F3 myoblasts enhanced their differentiation, as assessed by both morphological and biochemical criteria. Furthermore, expression of antisense CD164 or soluble extracellular regions of CD164 inhibited myogenic differentiation. Treatment of C2C12 cells with sialidase or O-sialoglycoprotease, two enzymes previously reported to destroy functional epitopes on CD164, also inhibited differentiation. These data indicate that (i) CD164 may play a rate-limiting role in differentiation of cultured myoblasts, (ii) sialomucins represent a class of potential effectors of cell contact-mediated regulation of myogenesis, and (iii) carbohydrate-based cell recognition may play a role in mediating this phenomenon.


Assuntos
Antígenos CD , Glicoproteínas de Membrana/fisiologia , Mucinas/fisiologia , Músculos/citologia , Moléculas de Adesão de Célula Nervosa , Receptores de Superfície Celular/fisiologia , Animais , Antígeno CD146 , Diferenciação Celular , Linhagem Celular , Endolina , Fibroblastos/citologia , Glicoproteínas de Membrana/genética , Metaloendopeptidases/metabolismo , Camundongos , Mucinas/genética , Mucolipidoses/metabolismo , Neuraminidase/metabolismo , Oligonucleotídeos Antissenso , Receptores de Superfície Celular/genética , Saccharomyces cerevisiae , Sialomucinas , Solubilidade , Fatores de Transcrição , Regulação para Cima
10.
Arch Pharm Res ; 24(4): 276-80, 2001 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-11534756

RESUMO

Substituted isoquinolin-1-ones (1) were synthesized to test their in vitro anticancer activity. 3-Biphenyl-N-methylisoquinolin-1-one (7) showed the most potent anticancer activity against five different human cancer cell lines.


Assuntos
Antineoplásicos/síntese química , Antineoplásicos/farmacologia , Isoquinolinas/síntese química , Isoquinolinas/farmacologia , Ensaios de Seleção de Medicamentos Antitumorais , Humanos , Indicadores e Reagentes , Espectroscopia de Ressonância Magnética , Relação Estrutura-Atividade , Células Tumorais Cultivadas
11.
Arch Pharm Res ; 24(1): 74-8, 2001 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-11235816

RESUMO

A preparation of water soluble components (EA) was made from carpophores of Elfvingia applanata (Pers.) Karst and its in vitro antiviral activity on vesicular stomatitis virus [(Indiana serotype, VSV(IND)] was investigated by plaque reduction assay. EA exhibited potent antiviral activity on VSV(IND) growth and negligible cytotoxicity on Vero cells, 50% effective concentration (EC50) of 104 microg/ml and 50% cytotoxic concentration (CC50) of 3,793 microg/ml, respectively. Selectivity index (SI, CC50/EC50) of EA on Vero cell and VSV(IND) was about 36.5. EA did not display either a direct virucidal effect on VSV(IND) or induction of antiviral substance by Vero cells upon its treatment. Thus, the mode of antiviral activity of EA was studied at steps of viral adsorption onto cell. When both EA and virus were added to cell monolayers, titer of cell-free virus in culture supernatant increased in ca. 30-40% compared with that of control group and titer of cell-associated virus was 60-100% higher than that of control group. These results suggested that antiviral activity of EA on VSV(IND) might be due to the hindrance of viral entry to cells at either endocytosis or loss of envelope.


Assuntos
Antivirais/farmacologia , Endocitose/efeitos dos fármacos , Polyporaceae , Vírus da Estomatite Vesicular Indiana/efeitos dos fármacos , Proteínas do Envelope Viral/efeitos dos fármacos , Animais , Chlorocebus aethiops , Endocitose/fisiologia , Interferon-alfa/farmacologia , Polyporaceae/química , Solubilidade , Células Vero , Vírus da Estomatite Vesicular Indiana/fisiologia , Proteínas do Envelope Viral/fisiologia
12.
Mol Cell Biochem ; 212(1-2): 29-34, 2000 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-11108133

RESUMO

Nucleic acid molecules with high affinities for a target transcription factor can be introduced into cells as decoy cis-elements to bind these factors and alter gene expression. This review discusses a synthetic single-stranded palindromic oligonucleotide, which self-hybridizes to form a duplex/hairpin and competes with cAMP response element (CRE) enhancers for binding transcription factors. This oligonucleotide inhibits CRE- and Ap-1-directed gene transcription and promotes growth inhibition in vitro and in vivo in a broad spectrum of cancer cells, without adversely affecting normal cell growth. Evidence presented here suggests that the CRE-decoy oligonucleotide can provide a powerful new means of combating cancers, viral diseases, and other pathological conditions by regulating the expression of cAMP-responsive genes.


Assuntos
Divisão Celular/efeitos dos fármacos , Proteína de Ligação ao Elemento de Resposta ao AMP Cíclico/metabolismo , Regulação da Expressão Gênica/efeitos dos fármacos , Oligodesoxirribonucleotídeos/farmacologia , Transcrição Gênica/efeitos dos fármacos , Animais , Elementos Facilitadores Genéticos , Genes fos , Humanos , Oligodesoxirribonucleotídeos/uso terapêutico , Fator de Transcrição AP-1/metabolismo , Células Tumorais Cultivadas , Proteína Supressora de Tumor p53/metabolismo
13.
Biochem Biophys Res Commun ; 278(3): 679-84, 2000 Nov 30.
Artigo em Inglês | MEDLINE | ID: mdl-11095968

RESUMO

In a recent report (Cho et al., Proc. Natl. Acad. Sci. USA 97, 835-840, 2000), we showed that cancer cells of various cell types secrete cAMP-dependent protein kinase (PKA) into the conditioned medium and that in the serum of cancer patients this extracellular PKA (ECPKA) is upregulated 10-fold as compared with normal serum. Here, we characterized the enzymatic properties of ECPKA that is present in the conditioned medium of PC3M prostate cancer cells and in the serum of cancer patients, and we compared ECPKA with PKA found in the cell extracts of PC3M cells. ECPKA present in the conditioned medium and human serum was not activated by cAMP addition, but intracellular PKA activity was totally dependent on the addition of cAMP. This indicates that the ECPKA is present in active, free C subunit form, whereas intracellular PKA is present in inactive holoenzyme form. ECPKA activity increased in a substrate concentration- and time-dependent manner, as did intracellular PKA. Both ECPKA and intracellular PKA activities were specifically inhibited by the PKA inhibitor protein, PKI. However, ECPKA activity was more temperature-sensitive than intracellular PKA; after two cycles of freezing/thawing, only 20% of initial ECPKA activity was detected compared with over 40% of intracellular PKA activity. Western blot analysis revealed the presence of a 40 kDa C(alpha) subunit of PKA in both conditioned medium and in the serum of cancer patients. These results suggest that ECPKA, out of the context of cAMP regulation, may function as a growth factor promoting cell growth and transformation; thus, it may serve as a tumor biomarker.


Assuntos
Biomarcadores Tumorais/sangue , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Neoplasias/sangue , Meios de Cultivo Condicionados , AMP Cíclico/metabolismo , Proteínas Quinases Dependentes de AMP Cíclico/sangue , Proteínas Quinases Dependentes de AMP Cíclico/química , Humanos , Cinética , Masculino , Neoplasias/enzimologia , Oligopeptídeos/farmacologia , Neoplasias da Próstata , Subunidades Proteicas , Células Tumorais Cultivadas
14.
Clin Cancer Res ; 6(9): 3434-41, 2000 Sep.
Artigo em Inglês | MEDLINE | ID: mdl-10999726

RESUMO

The cyclic AMP-dependent protein kinase (PKA) exists in two isoforms, PKA-I (type I) and PKA-II (type II), that contain an identical catalytic (C) subunit but distinct regulatory (R) subunits, RI and RII, respectively. Increased expression of RIalpha/PKA-I has been shown in human cancer cell lines, in primary tumors, in cells after transformation, and in cells upon stimulation of growth. We have shown previously that a single-injection RI, antisense treatment results in a reduction in RIalpha and PKA-I expression and sustained inhibition of human colon carcinoma growth in athymic mice (M. Nesterova and Y. S. Cho-Chung, Nat. Med., 1: 528-533, 1995). Growth inhibition accompanied reduction in RIalpha/PKA-I expression and compensatory increases in RIIbeta protein and PKA-IIbeta, the RIIbeta-containing holoenzyme. Here, we report that these in vivo findings are consistent with observations made in cancer cells in culture. We demonstrate that the antisense depletion of RIalpha in cancer cells results in increased RIIbeta protein without increasing the rate of RIIbeta synthesis or RIIbeta mRNA levels. Pulse-chase experiments revealed a 3-6-fold increase in the half-life of RIIbeta protein in antisense-treated colon and prostate carcinoma cells with little or no change in the half-lives of RIalpha, RIIalpha, and Calpha proteins. Compensation by RIIbeta stabilization may represent a novel biochemical adaptation mechanism of the cell in response to sequence-specific loss of RIalpha expression, which leads to sustained down-regulation of PKA-I activity and inhibition of tumor growth.


Assuntos
Neoplasias do Colo/enzimologia , Proteínas Quinases Dependentes de AMP Cíclico/antagonistas & inibidores , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Oligonucleotídeos Antissenso/farmacologia , Neoplasias da Próstata/enzimologia , Northern Blotting , Western Blotting , Ciclo Celular/efeitos dos fármacos , Ciclo Celular/fisiologia , Divisão Celular/efeitos dos fármacos , Divisão Celular/fisiologia , Neoplasias do Colo/genética , Neoplasias do Colo/patologia , Subunidade RIIbeta da Proteína Quinase Dependente de AMP Cíclico , Subunidade RIalfa da Proteína Quinase Dependente de AMP Cíclico , Proteína Quinase Tipo II Dependente de AMP Cíclico , Proteínas Quinases Dependentes de AMP Cíclico/biossíntese , Proteínas Quinases Dependentes de AMP Cíclico/genética , Estabilidade Enzimática , Regulação Enzimológica da Expressão Gênica/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Inibidores do Crescimento/farmacologia , Humanos , Masculino , Oligonucleotídeos Antissenso/genética , Neoplasias da Próstata/genética , Neoplasias da Próstata/patologia , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fase S/efeitos dos fármacos , Fase S/fisiologia , Especificidade por Substrato , Tionucleotídeos/genética , Tionucleotídeos/farmacologia , Células Tumorais Cultivadas , Regulação para Cima/efeitos dos fármacos
15.
Biochemistry ; 39(16): 4863-8, 2000 Apr 25.
Artigo em Inglês | MEDLINE | ID: mdl-10769144

RESUMO

The CRE, 5'-TGACGTCA-3', has been described as the consensus sequence for the cis-element that directs cAMP-regulated gene expression. Many transcription factors bind to this element and regulate the expression of a wide variety of cellular and viral genes. We have shown that CRE-transcription factor decoy oligonucleotide restrains the growth of cancer cells in vitro and in vivo [Park, Y. G., Nesterova, M., Agrawal, S., and Cho-Chung, Y. S. (1999) J. Biol. Chem. 274, 1573-1580]. The growth inhibition was accompanied by changes in cell morphology and apoptosis. To elucidate the molecular mechanism(s) of the growth inhibition by the CRE-decoy oligonucleotide, we investigated the p53 signaling pathway. Herein, we report that CRE-decoy oligonucleotide treatment results in an increase in the p53 protein level in MCF-7 human breast cancer cells that express wild-type p53. The p21WAF1/Cip1 protein levels were also increased in the CRE-decoy oligonucleotide treated cells accompanying a reduction in Cdk2- and cyclin E-dependent kinase activity and pRb phosphorylation. Pulse-chase experiments reveal that the p53 upregulation was due to increased stability of the protein. The decoy oligonucleotide treatment also enhanced the p53 promotor-directed transcription in vivo along with the increase in p53-CBP (CREB-binding protein) complex formation. Thus, the stabilization and activation of p53 may have contributed to the growth inhibition induced by CRE-transcription factor decoy oligonucleotide in MCF-7 breast cancer cells. This decoy oligonucleotide approach offers great promise as a tool for defining cellular regulatory processes and treating cancer and other diseases.


Assuntos
Neoplasias da Mama/patologia , Quinases relacionadas a CDC2 e CDC28 , AMP Cíclico/fisiologia , Oligonucleotídeos/farmacologia , Elementos de Resposta/genética , Transdução de Sinais/efeitos dos fármacos , Proteína Supressora de Tumor p53/metabolismo , Neoplasias da Mama/tratamento farmacológico , Neoplasias da Mama/enzimologia , Neoplasias da Mama/metabolismo , Proteína de Ligação a CREB , Divisão Celular/efeitos dos fármacos , Ciclina E/metabolismo , Quinase 2 Dependente de Ciclina , Inibidor de Quinase Dependente de Ciclina p21 , Quinases Ciclina-Dependentes/metabolismo , Ciclinas/metabolismo , Regulação para Baixo/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Proteínas Nucleares/metabolismo , Oligonucleotídeos/genética , Oligonucleotídeos/uso terapêutico , Fosforilação/efeitos dos fármacos , Regiões Promotoras Genéticas/genética , Ligação Proteica/efeitos dos fármacos , Proteínas Serina-Treonina Quinases/metabolismo , Proteína do Retinoblastoma/metabolismo , Termodinâmica , Transativadores/metabolismo , Ativação Transcricional/efeitos dos fármacos , Células Tumorais Cultivadas , Proteína Supressora de Tumor p53/genética
16.
Proc Natl Acad Sci U S A ; 97(2): 835-40, 2000 Jan 18.
Artigo em Inglês | MEDLINE | ID: mdl-10639166

RESUMO

Overexpression of cAMP-dependent protein kinase (PKA) type I isozyme is associated with cell proliferation and neoplastic transformation. The presence of PKA on the external surface of LS-174T human colon carcinoma cells has been shown. Here, we show that cancer cells of various cell types excrete PKA into the conditioned medium. This extracellular PKA (ECPKA) is present in active, free catalytic subunit (C subunit) form, and its activity is specifically inhibited by PKA inhibitory protein, PKI. Overexpression of the Calpha or RIalpha subunit gene of PKA in an expression vector, which up-regulates intracellular PKA type I, markedly up-regulates ECPKA expression. In contrast, overexpression of the RIIbeta subunit, which eliminates PKA type I, up-regulates PKA type II, and reverts the transformed phenotype, down-regulates ECPKA. A mutation in the Calpha gene that prevents myristylation allows the intracellular PKA up-regulation but blocks the ECPKA increase, suggesting that the NH(2)-terminal myristyl group of Calpha is required for the ECPKA expression. In serum of cancer patients, the ECPKA expression is up-regulated 10-fold as compared with normal serum. These results indicate that the ECPKA expression is an ordered cellular response of a living cell to actively exclude excess intracellular PKA molecules from the cell. This phenomenon is up-regulated in tumor cells and has an inverse relationship with the hormone dependency of breast cancer. Thus, the extracellular PKA may serve as a potential diagnostic and prognostic marker for cancer.


Assuntos
Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , Células 3T3/citologia , Células 3T3/enzimologia , Animais , Biomarcadores Tumorais/análise , Biomarcadores Tumorais/metabolismo , Neoplasias da Mama/enzimologia , Neoplasias da Mama/patologia , Linhagem Celular , Transformação Celular Neoplásica , Meios de Cultivo Condicionados , AMP Cíclico/farmacologia , Proteína Quinase Tipo II Dependente de AMP Cíclico , Proteínas Quinases Dependentes de AMP Cíclico/análise , Proteínas Quinases Dependentes de AMP Cíclico/genética , Espaço Extracelular/enzimologia , Regulação Enzimológica da Expressão Gênica , Hormônios/farmacologia , Humanos , Isoenzimas/genética , Isoenzimas/metabolismo , Camundongos , Mutação , Ácido Mirístico/metabolismo , Neoplasias/sangue , Neoplasias/enzimologia , Antígeno Prostático Específico/biossíntese , Proteínas Recombinantes de Fusão/análise , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo , Células Tumorais Cultivadas/enzimologia
17.
Front Biosci ; 4: D898-907, 1999 Dec 01.
Artigo em Inglês | MEDLINE | ID: mdl-10577386

RESUMO

Enhanced expression of the RIa subunit of cAMP-dependent protein kinase type I (PKA-I) has been shown during carcinogenesis, in human cancer cell lines and in primary tumors. We demonstrate that the sequence-specific inhibition of RIa gene expression by antisense oligonucleotides results in the differentiation of leukemia cells and growth arrest of cancer cells of epithelial origin and tumors in mice. The loss of RI by the antisense results in rapid increase in the half-life of the competitor molecule, RII protein, via its stabilization in a holoenzyme complex (PKA-II) that insures depletion of PKA-I and sustained inhibition of tumor growth. RI antisense, which restrains tumor cell growth by turning on the signals for blockade of tumor cell survival, namely blockade of the tyrosine kinase signaling, cell cycle deregulation and apoptosis, provides a single gene-targeting approach to treatment of cancer.


Assuntos
Proteínas Quinases Dependentes de AMP Cíclico/genética , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , DNA Antissenso/uso terapêutico , Neoplasias/terapia , Animais , Apoptose , Ciclo Celular/genética , Diferenciação Celular/genética , Divisão Celular/genética , Subunidade RIIalfa da Proteína Quinase Dependente de AMP Cíclico , Subunidade RIIbeta da Proteína Quinase Dependente de AMP Cíclico , Subunidade RIalfa da Proteína Quinase Dependente de AMP Cíclico , Proteínas Quinases Dependentes de AMP Cíclico/biossíntese , Proteínas Quinases Dependentes de AMP Cíclico/fisiologia , DNA Antissenso/genética , Resistencia a Medicamentos Antineoplásicos/genética , Humanos , Camundongos , Camundongos Nus , Neoplasias/genética , Neoplasias/metabolismo , Neoplasias/patologia , Neoplasias/prevenção & controle , Neoplasias Experimentais/genética , Neoplasias Experimentais/metabolismo , Neoplasias Experimentais/patologia , Neoplasias Experimentais/prevenção & controle , Neoplasias Experimentais/terapia , Proteínas Tirosina Quinases/antagonistas & inibidores , RNA Mensageiro/biossíntese , Células Tumorais Cultivadas
18.
Curr Opin Mol Ther ; 1(3): 386-92, 1999 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-11713803

RESUMO

Cellular and molecular research has been focused to develop a means to regulate gene expression in an effort to treat and cure a variety of diseases and abnormal physiological conditions. A successful oligonucleotide-based approach has been the use of synthetic oligonucleotides containing an enhancer element that can penetrate cells, bind sequence-specific DNA-binding proteins and interfere with transcription in vivo. This review describes such decoy oligonucleotides that exhibit high affinity for a target transcription factor and successfully interfere with transcription in vivo. Evidence presented here shows that the decoy oligonucleotide technology offers great promise as a tool for defining cellular regulatory processes and for treating cancer, viral diseases and other pathological conditions.


Assuntos
Proteínas de Ciclo Celular , Proteínas de Ligação a DNA , Regulação da Expressão Gênica/efeitos dos fármacos , Oligodesoxirribonucleotídeos/farmacologia , Tionucleotídeos/farmacologia , Fatores de Transcrição/antagonistas & inibidores , Angioplastia Coronária com Balão , Adesão Celular/efeitos dos fármacos , Núcleo Celular/química , Estenose Coronária/prevenção & controle , Estenose Coronária/terapia , AMP Cíclico/fisiologia , DNA/metabolismo , Desenho de Fármacos , Estabilidade de Medicamentos , Fatores de Transcrição E2F , Elementos Facilitadores Genéticos/genética , Produtos do Gene tat/antagonistas & inibidores , Produtos do Gene tat/genética , Genes rev , Genes tat , Glomerulonefrite/tratamento farmacológico , Glomerulonefrite/genética , Transcriptase Reversa do HIV/genética , Infarto do Miocárdio/tratamento farmacológico , Infarto do Miocárdio/genética , NF-kappa B/antagonistas & inibidores , NF-kappa B/fisiologia , Conformação de Ácido Nucleico , Oligodesoxirribonucleotídeos/química , Regiões Promotoras Genéticas/efeitos dos fármacos , Ligação Proteica/efeitos dos fármacos , Recidiva , Sequências Reguladoras de Ácido Nucleico , Inibidores da Transcriptase Reversa/química , Inibidores da Transcriptase Reversa/farmacologia , Sistemas do Segundo Mensageiro/fisiologia , Relação Estrutura-Atividade , Especificidade por Substrato , Acetato de Tetradecanoilforbol/farmacologia , Túnica Íntima/efeitos dos fármacos , Túnica Íntima/metabolismo , Túnica Íntima/patologia
19.
Proc Natl Acad Sci U S A ; 95(12): 6687-92, 1998 Jun 09.
Artigo em Inglês | MEDLINE | ID: mdl-9618473

RESUMO

cAMP, through the activation of cAMP-dependent protein kinase (PKA), is involved in transcriptional regulation. In eukaryotic cells, cAMP is not considered to alter the binding affinity of CREB/ATF to cAMP-responsive element (CRE) but to induce serine phosphorylation and consequent increase in transcriptional activity. In contrast, in prokaryotic cells, cAMP enhances the DNA binding of the catabolite repressor protein to regulate the transcription of several operons. The structural similarity of the cAMP binding sites in catabolite repressor protein and regulatory subunit of PKA type II (RII) suggested the possibility of a similar role for RII in eukaryotic gene regulation. Herein we report that RIIbeta subunit of PKA is a transcription factor capable of interacting physically and functionally with a CRE. In contrast to CREB/ATF, the binding of RIIbeta to a CRE was enhanced by cAMP, and in addition, RIIbeta exhibited transcriptional activity as a Gal4-RIIbeta fusion protein. These experiments identify RIIbeta as a component of an alternative pathway for regulation of CRE-directed transcription in eukaryotic cells.


Assuntos
Proteínas Quinases Dependentes de AMP Cíclico/química , Proteínas Quinases Dependentes de AMP Cíclico/metabolismo , AMP Cíclico/metabolismo , Transdução de Sinais , Células 3T3 , Animais , Sítios de Ligação , AMP Cíclico/química , Subunidade RIIbeta da Proteína Quinase Dependente de AMP Cíclico , Proteína Quinase Tipo II Dependente de AMP Cíclico , Proteínas Quinases Dependentes de AMP Cíclico/genética , Camundongos , Proteínas Recombinantes de Fusão/química , Proteínas Recombinantes de Fusão/genética , Proteínas Recombinantes de Fusão/metabolismo
20.
J Steroid Biochem Mol Biol ; 67(2): 105-11, 1998 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-9877210

RESUMO

We have previously reported that ginsenosides Rh1 and Rh2 induced the differentiation of F9 teratocarcinoma stem cells [Lee, Y. N., Lee, H. Y., Chung, H. Y., Kim, S. I., Lee, S. K., Park, B. C. and Kim, K. W., In vitro induction of differentiation by ginsenosides in F9 teratocarcinoma cells. Eur. J. Cancer 1996, 32, 1420-1428.]. Since the chemical structure of Rh1 and Rh2 is very similar to that of dexamethasone, a synthetic glucocorticoid, we investigated whether Rh1 and Rh2 act through the glucocorticoid receptor (GR). Immunocytochemistry showed that Rh1 or Rh2 increased the nuclear translocation of GR in the same manner of dexamethasone. In the gel shift assay, glucocorticoid response element (GRE) binding protein in F9 cells was increased by Rh1 or Rh2. To confirm whether the increased binding protein is GR, we performed the competition assay with unlabeled GRE as a specific competitor. Moreover, supershift assay with the GR antibody showed that the binding proteins are GR. In addition, to confirm the Rh1 or Rh2-induced transactivation of GRE promoter, we cotransfected GR expression vector and GRE-luciferase vector. In the luciferase assay, Rh1 or Rh2 potently induced luciferase activity and this induction was blocked by RU486, a potent GR antagonist. Taken together, we suggest that ginsenosides Rh1 and Rh2 may induce the differentiation of F9 cells by stimulating the nuclear translocation of GR.


Assuntos
Fármacos do Sistema Nervoso Central/farmacologia , Receptores de Glucocorticoides/metabolismo , Saponinas/farmacologia , Transdução de Sinais/efeitos dos fármacos , Teratocarcinoma/metabolismo , Teratocarcinoma/patologia , Animais , Diferenciação Celular/efeitos dos fármacos , Ginsenosídeos , Camundongos , Células Tumorais Cultivadas
SELEÇÃO DE REFERÊNCIAS
DETALHE DA PESQUISA