RESUMO
Abstract Anterior open bite (AOB) has a multifactorial etiology caused by the interaction of sucking habits and genetic factors. The aim of this study was to evaluate the association between AOB and polymorphisms in genes that encode Matrix metalloproteinases (MMPs) and their tissue inhibitors (TIMPs). Four hundred and seventy-two children that presented at least one sucking habit were evaluated. Children were examined clinically for the presence of AOB. Genomic DNA was extracted from saliva. Genotyping of the selected polymorphisms in MMP2, MMP3, MMP9, TIMP1 and TIMP2 was carried out by real-time PCR using the TaqMan method. Allele and genotype frequencies were compared between the groups with and without AOB using the PLINK® software in a free and in a recessive model using a chi-square test. Logistic regression analysis was implemented (p≤0.05). Two hundred nineteen children had AOB while 253 did not. The polymorphism rs17576 in MMP9 was significantly associated with AOB (p=0.009). In a recessive model GG genotype was a protective factor for AOB (p=0.014; OR 4.6, 95%CI 1.3-16.2). In the logistic regression analysis, none of the genes was associated with AOB. In conclusion, the polymorphism rs17576 (glutamine for arginine substitution) in MMP9 was a protective factor for AOB.
Resumo A mordida aberta anterior apresenta uma etiologia multifatorial causada pela interação entre hábitos de sucção e fatores genéticos. O objetivo deste estudo foi avaliar a associação entre mordida aberta anterior e polimorfismo nos genes que codificam as metaloproteinases da matriz (MMPs) e seus inibidores teciduais (TIMPs). Foram avaliadas 472 crianças que apresentvam pelo menos um hábito de sucção. As crianças foram clinicamente examinadas para avaliar a presença de mordida aberta anterior. DNA genômico foi extraído da saliva. A genotipagem dos polimorfismos selecionados em MMP2, MMP3, MMP9, TIMP1 e TIMP2 foi realizada por PCR em tempo real, usando o método de TaqMan. As frequências alélicas e genotípicas foram comparadas entre os grupos com e sem mordida aberta anterior usando o software PLINK®. Duzentas e dezenove crianças apresentavam mordida aberta anterior enquanto 253 não a apresentavam. O polimorfismo rs17576 em MMP9 estava significativamente associado com mordida aberta anterior (p=0,009). No modelo recessivo (GG versus AG+AA) o genótipo GG foi um fator protetor para mordida aberta anterior (p=0,014; OR 4,6; 95%CI 1,3- 16,2). Concluindo, o polimorfismo rs17576 (substituição de glutamina por arginina) em MMP9 está associado com mordida aberta anterior. Os resultados obtidos suportam a hipótese de que fatores genéticos estão envolvidos com a etiologia da mordida aberta anterior.
Assuntos
Humanos , Masculino , Feminino , Pré-Escolar , Criança , Metaloproteinase 9 da Matriz/genética , Polimorfismo de Nucleotídeo Único , Mordida Aberta/etiologia , Metaloproteinase 3 da Matriz/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-2/genética , Metaloproteinase 2 da Matriz/genética , Mordida Aberta/genética , Reação em Cadeia da Polimerase em Tempo Real , Sucção de Dedo , Frequência do Gene , Genótipo , Modelos GenéticosRESUMO
The Brazilian Amazon Region is a highly endemic area for hepatitis B virus (HBV). However, little is known regarding the genetic variability of the strains circulating in this geographical region. Here, we describe the first full-length genomes of HBV isolated in the Brazilian Amazon Region; these genomes are also the first complete HBV subgenotype D3 genomes reported for Brazil. The genomes of the five Brazilian isolates were all 3,182 base pairs in length and the isolates were classified as belonging to subgenotype D3, subtypes ayw2 (n = 3) and ayw3 (n = 2). Phylogenetic analysis suggested that the Brazilian sequences are not likely to be closely related to European D3 sequences. Such results will contribute to further epidemiological and evolutionary studies of HBV.
Assuntos
Humanos , Carcinoma Hepatocelular , Movimento Celular/fisiologia , Neoplasias Hepáticas , Metaloproteinase 9 da Matriz/genética , Transdução de Sinais/fisiologia , Inibidor Tecidual de Metaloproteinase-1/genética , Linhagem Celular Tumoral/citologia , Linhagem Celular Tumoral/fisiologia , Colagenases/genética , Dipeptídeos/farmacologia , Inibidores de Metaloproteinases de Matriz , Metaloproteinase 1 da Matriz/genética , /genética , /genética , /genética , Inibidores de Proteases/farmacologia , Transdução de Sinais/efeitos dos fármacos , TransfecçãoRESUMO
O colesteatoma adquirido da orelha média é uma lesão epitelial escamosa queratinizante e hiperproliferativa benigna que pode resultar na destruição das estruturas ósseas circunvizinhas do osso temporal. Estudos recentes demonstram que alterações na produção celular de metaloproteinases de matriz (MMPs) e seus inibidores específicos (TIMPs) contribuem para a fisiopatologia do colesteatoma. OBJETIVO: Verificar a aplicabilidade da amplificação de RNA para avaliação da expressão de isoformas de MMPs e TIMPS em colesteatomas para correlação com a agressividade da doença. MATERIAIS E MÉTODOS: Estudo prospectivo. Dezenove casos de colesteatomas em diferentes estágios de evolução foram selecionados. RNA extraído das biópsias foi submetido à transcrição reversa - reação da polimerase em cadeia (RT-PCR) para amplificação semiquantitativa de MMP2, MMP3, MMP9, MMP13 e TIMP1. Resultados: Seis colesteatomas apresentaram reação positiva para pelo menos um dos genes estudados. Quatro amostras amplificaram apenas um gene (MMP2 ou MM13) e duas amostras amplificaram três genes (MMP2, TIMP1 e MMP3 ou MMP13). Nenhuma amostra amplificou MMP9. CONCLUSÃO: A avaliação da expressão gênica de MMPs e TIMPs em colesteatomas pode ser realizada por RT-PCR, apesar de dificuldades técnicas. Não foi possível realizar associação entre o perfil de expressão gênica e a agressividade da doença.
Acquired middle ear cholesteatoma is a benign keratinizing hyperproliferative squamous epithelial lesion that may result in the destruction of the bone structures surrounding the temporal bone. Recent studies show that variations in cellular production of matrix metalloproteinases (MMPs) and their specific inhibitors (TIMPs) contribute to the pathophysiology of cholesteatoma. OBJECTIVE: This study aims to analyze the use of RNA amplification tests to evaluate the expression of MMP and TIMP isoforms in cholesteatomas and their correlation with disease severity. MATERIALS AND METHODS: This is a prospective study. Nineteen cholesteatoma cases at different stages were selected. RNA collected from biopsy specimens was submitted to reverse transcription polymerase chain reaction (RT-PCR) for semiquantitative amplification of MMP2, MMP3, MMP9, MMP13 and TIMP1. RESULTS: Six cholesteatomas were positive for at least one of the studied genes. Four samples amplified a single gene (MMP2 or MMP13) and two samples amplified three genes (MMP2, TIMP1 and MMP3 or MMP13). No sample amplified MMP9. CONCLUSION: RT-PCR can be used to assess MMP and TIMP gene expression in cholesteatomas despite technical difficulties. Gene expression profiles could not be related to disease severity.
Assuntos
Adolescente , Adulto , Idoso , Criança , Pré-Escolar , Humanos , Pessoa de Meia-Idade , Adulto Jovem , Colesteatoma da Orelha Média/enzimologia , Expressão Gênica/genética , Metaloproteinases da Matriz/genética , RNA Mensageiro/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Audiometria , Biomarcadores/análise , Colesteatoma da Orelha Média/complicações , Colesteatoma da Orelha Média/genética , Perda Auditiva/diagnóstico , Perda Auditiva/etiologia , Metaloproteinases da Matriz/metabolismo , Estudos Prospectivos , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Mensageiro/metabolismo , Índice de Gravidade de Doença , Tomografia Computadorizada por Raios X , Inibidor Tecidual de Metaloproteinase-1/metabolismoRESUMO
The study investigated the effects of adenovirus-mediated gene transfection of basic fibroblast growth factor (bFGF), bFGF combined with interleukin-1 receptor antagonist protein (IL-Ra) and/or insulin-like growth factor-1 (IGF-1) both in human osteoarthritis (OA) chondrocytes and rabbits OA model. Human OA chondrocytes were delivered by adenovirus-mediated bFGF, IL-Ra and IGF-1 vectors, respectively. Chondrocyte proliferation, glycosaminoglycan (GAG) content, expression of type II collagen, ADAMTS-5, MMP-13, MMP-3 and TIMP-1 were determined. Rabbit OA model was induced by anterior cruciate ligament transaction (ACLT) in knees. Adenoviral vectors encoding human bFGF, IL-Ra and IGF-1 were injected intraarticularly into the knee joints after ACLT. The effects of adenovirus- mediated gene transfection on rabbit OA were evaluated. In vitro, the transfected genes were expressed in cell supernatant of human OA chondrocytes. AdbFGF group significantly promoted chondrocyte proliferation, and increased GAG and type II collagen synthesis than in the OA group. As two or three genes were transfected in different combinations, there was significant enhancement on the GAG content, type II collagen synthesis, and TIMP-1 levels, while ADAMTS-5, MMP-13, and MMP-3 levels were reduced. In vivo, the transfected genes were expressed in synovial fluid of rabbits. Intraarticular delivery of bFGF enhanced the expression of type II collagen in cartilage and decreased cartilage Mankin score compared with the OA control group (P = 0.047; P < 0.01, respectively). Multiple-gene transfection in different combinations showed better results than bFGF transfection alone. This study suggests that bFGF gene transfection is effective in treating experimental OA. Multiple gene transfection has better biologic effects on OA.
Assuntos
Animais , Humanos , Coelhos , Adenoviridae/genética , Condrócitos/efeitos dos fármacos , Colágeno Tipo II/genética , Fator 2 de Crescimento de Fibroblastos/genética , Terapia Genética/métodos , Vetores Genéticos/administração & dosagem , Fator de Crescimento Insulin-Like I/genética , Proteína Antagonista do Receptor de Interleucina 1/genética , Interleucina-1/genética , Metaloproteinase 13 da Matriz/genética , Metaloproteinase 3 da Matriz/genética , Osteoartrite/terapia , Inibidor Tecidual de Metaloproteinase-1/genética , TransfecçãoRESUMO
Background: Several molecules that may have a role in tumor proliferation, differentiation and invasion, have been detected in thyroid carcinoma. Some of these molecules are NIS, c-MET, TIMP1 an ephrinB2. Aim: To detect the presence of these molecules in tissue samples of thyroid carcinoma and relate their expression to the biological behavior of the tumor. Material and Methods: Tissue samples were prospectively obtained from 35 patients operated for a papillary thyroid carcinoma. Twelve patients had regional lymph node involvement. NIS, c-MET, TIMP1 and EphrinB2 were detected by real time polymerase chain reaction(RT-PCR) and immunohistochemistry. Results: The expression of markers by RT-PCR was non significantly higher among tumors with lymph node involvement. Immunohistochemistryshowed a significantly lower nuclear expression and a higher cytoplasmatic expression of EphrinB2 in tumors with lymph node involvement. Conclusions: Immunohistochemical expression of EphrinB2 could be useful for the initial staging of papillary thyroid carcinoma.
Assuntos
Humanos , Adenocarcinoma Papilar/genética , Adenocarcinoma Papilar/metabolismo , Neoplasias da Glândula Tireoide/genética , Neoplasias da Glândula Tireoide/metabolismo , /genética , /metabolismo , Regulação Neoplásica da Expressão Gênica , Imuno-Histoquímica , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Metástase Linfática , Biomarcadores Tumorais , Invasividade Neoplásica , Proteínas Proto-Oncogênicas c-met/genética , Proteínas Proto-Oncogênicas c-met/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Simportadores/metabolismoRESUMO
The tissue inhibitor of metalloproteinases (TIMP)-1 is a multifunctional protein which is not only an inhibitor of matrix metalloproteinases (MMPs) but also to have a possible "cytokine-like" action. Here, we first compared mRNA expression of TIMP-1 and MMP-9 in BEL-7402 (a hepatocellular carcinoma cell line), L-02 (a normal liver cell line) and QSG-7701 (a cell line derived from peripheral tissue of liver carcinoma) using real-time quantitative RT-PCR. By evaluating the variation of the MMP-9/TIMP-1 ratio as an index of reciprocal changes of the expression of the two genes, we observed that the MMP-9/TIMP-1 ratio was about 13- and 5-fold higher in BEL-7402 than in L-02 and QSG-7701, respectively. Significantly, overexpression of TIMP-1 decreased the MMP-9/TIMP-1 ratio in BEL-7402 and then inhibited the cell growth to 60 percent and reduced the migration to about 30 percent. Meanwhile, our data showed that interleukin-6 (IL-6) (100 ng/mL) could also inhibited the cell growth of BEL-7402. Further studies indicated that TIMP-1 mediated the inhibitory effect of IL-6 on BEL-7402 cell proliferation in a STAT3-dependent manner, which could further accelerate the expression of the cyclin-dependent kinase inhibitor p21. A dominant negative STAT3 mutant totally abolished IL-6-induced TIMP-1 expression and its biological functions. The present results demonstrate that TIMP-1 may be one of the mediators that regulate the inhibitory effect of IL-6 on BEL-7402 proliferation in which STAT3 signal transduction and p21 up-regulation also play important roles.
Assuntos
Humanos , Carcinoma Hepatocelular/genética , /genética , Neoplasias Hepáticas/genética , Metaloproteinase 9 da Matriz/genética , /genética , Inibidor Tecidual de Metaloproteinase-1/genética , Linhagem Celular Tumoral , Movimento Celular , Proliferação de Células , Carcinoma Hepatocelular/metabolismo , Carcinoma Hepatocelular/patologia , /metabolismo , Neoplasias Hepáticas/metabolismo , Neoplasias Hepáticas/patologia , Metaloproteinase 9 da Matriz/metabolismo , Reação em Cadeia da Polimerase Via Transcriptase Reversa , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Transdução de Sinais , /metabolismo , Inibidor Tecidual de Metaloproteinase-1/metabolismo , Regulação para CimaRESUMO
PURPOSE: To evaluate using transcription analysis the presence and importance of two genes: NM23-H1 and TIMP-1 on control of tumor cell invasion in diffuse astrocytomas (WHO II) and glioblastoma multiforme (WHO IV). METHOD: Northern blot analysis of NM23-H1 and TIMP-1 was performed. Eight diffuse astrocytomas and 19 glioblastomas (WHO IV) were analyzed to determine if TIMP-1 and NM23-H1 were candidates to inhibition of tumor cell invasion quantitated RNA levels. The samples were collected directly from operating room. Total cellular RNA was extracted from frozen tissue samples using guanidinium-isothiocyanate and cesium chloride gradients. Total RNA (10 mg per sample) from tumor tissue were size fractionated through 1 percent agarose-formaldehyde gel and transferred to nylon filters and then hybridized to 32P-labeled DNA probes and placed for autoradiography. Levels of specific RNAs were determined by computer-assisted laser densitometry. Blot filters were sequentially hybridized to nm23 and TIMP-1 probes in addition to GAPDH, as a control. Statistical analyses were carried out according to t-test for equality of means. RESULTS: NM23-H1 were detected in each sample, however it did not correlate with malignancy and invasiveness. On the other side TIMP-1 gene expression showed a clear correlation between low expression and invasiveness. CONCLUSION: The data suggest that TIMP-1 is an inhibitor of high grade gliomas invasion. NM23-H1 was present in the entire gliomas sample, but it did not vary in diffuse astrocytomas and glioblastomas.
OBJETIVO: Comparar através da análise da expressão dos níveis de RNA, a presença e a relevância dos genes NM23-H1 e TIMP-1 no controle da invasão celular tumoral dentro do tecido cerebral normal em: astrocitoma difuso (OMS II) e glioblastoma multiforme (OMS:IV). MÉTODO: Análise em "Northern blot" dos genes NM23-H1 e TIMP-1. Oito astrocitomas fibrilares difusos (OMS II) e 19 glioblastomas multiformes foram analisados para determinar se TIMP-1 e NM23-H1 estavam relacinados à inibição da invasão tumoral nas neoplasias do sistema nervoso central, quantificando os níveis de RNA dos respectivos genes extraídos diretamente dos tumores. 10 mg por amostra de RNA total foram fracionados de gel de formaldeído e transferidos para os filmes de hibridação. Níveis específicos de RNAs foram determinados na espectrofotometria. Valores das razões entre NM23-H1/GAPDH e TIMP-1/GAPDH foram submetidos à análise de variabilidade das médias. RESULTADOS: A análise da expressão do gene TIMP-1 mostrou supressão em tumores gliais malignos. CONCLUSÃO: Os resultados indicam que existe relação direta entre níveis baixos de TIMP-1 e malignidade dos gliomas. O gene NM23-H1 foi detectado em todas as amostras, mas não foi possível relacionar sua subexpressão ou superexpressão com algum fenótipo de invasividade.
Assuntos
Humanos , Astrocitoma/genética , Neoplasias Encefálicas/genética , Glioblastoma/genética , Proteínas de Neoplasias/genética , Núcleosídeo-Difosfato Quinase/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Northern Blotting , Regulação Neoplásica da Expressão Gênica , Marcadores Genéticos , Invasividade Neoplásica , RNA Neoplásico/análise , Transcrição GênicaRESUMO
Small hairpin RNA (shRNA) was used to silence the HIF1alpha gene in human retinal pigment epithelial cells (RPE) under hypoxia in order to observe the effect of gene silencing on the expression of matrix metalloproteinase tissue inhibitor 1 (TIMP1). By using chemical hypoxic inducer CoCl2, to mimic RPE hypoxic environment, shRNA against the targeting region of HIF1alpha mRNA sequence was synthesized by a method of in vitro transcription, and the HIF1alpha was interfered in RPE cultured under hypoxia (induced by 150 micromol/L CoCl2). RT-PCR was employed to detect the expression of HIF1alpha and TIMP1. The expression levels of HIF1alpha and TIMP1 were measured by using Western blotting. The results showed that after the RPE were transfected with specific shRNA against HIF1alpha mRNA, RT-PCR revealed that under hypoxia, the efficacy of HIF1alpha gene silencing in RPE was 83.4%. Western blotting revealed that the expression levels of HIF1alpha protein was dramatically dropped. In addition. RT-PCR results demonstrated that the expression of TIMP1 mRNA was decreased by 28.9%, and the expression levels of TIMP1 protein were also significantly reduced by Western blotting. It was suggested that shRNA targeted against HIF1alpha mRNA could effectively silence the HIF1alpha gene, subsequently effectively inhibit the hypoxia-induced up-regulation of TIMP1.
Assuntos
Hipóxia Celular , Células Cultivadas , Inativação Gênica , Subunidade alfa do Fator 1 Induzível por Hipóxia/genética , Epitélio Pigmentado Ocular/citologia , Epitélio Pigmentado Ocular/metabolismo , RNA Mensageiro/biossíntese , RNA Mensageiro/genética , RNA Interferente Pequeno/farmacologia , Retina/citologia , Retina/metabolismo , Inibidor Tecidual de Metaloproteinase-1/biossíntese , Inibidor Tecidual de Metaloproteinase-1/genéticaRESUMO
Biliary atresia is a progressive obliterative cholangiopathy, but the etiology of this disorder remains uncertain. Identifying genes specifically expressed in biliary atresia and analyzing the pattern of expression may lead to a better understanding of the pathogenesis. Liver tissues were taken from a recipient with biliary atresia and a normal donor during liver transplantation. Total RNA was extracted from each sample and reversely transcribed to cDNA. Then radiolabeled cDNA probe pools were made by random primed DNA labeling method and used for screening of differentially expressed genes by hybridizing with expressed sequence tags (EST) dot blot panel. Northern blot hybridization was done to confirm that these genes are also differentially expressed in other liver tissues. Among 1,730 EST clones, 26 cDNA clones were significantly overexpressed in biliary cirrhosis, while 2 clones were significantly decreased in biliary atresia. By Northern blot hybridization, the results of tissue inhibitor of metalloproteinase (TIMP)-1 and IGFBP-2 were well correlated with differential EST screening (DES). This study identified the pattern of differentially expressed genes in the biliary cirrhosis due to biliary atresia using DES technique.
Assuntos
Humanos , Atresia Biliar/genética , Northern Blotting , Perfilação da Expressão Gênica/métodos , Biblioteca Gênica , Proteína 2 de Ligação a Fator de Crescimento Semelhante à Insulina/genética , Inibidor Tecidual de Metaloproteinase-1/genéticaRESUMO
To evaluate possible roles of matrix metalloproteinase (MMP)-1, -2, tissue inhibitor of metalloproteinase (TIMP)-1, -2 and membrane-type-1 matrix metalloproteinase (MT1-MMP) in invasion of human gliomas, expressions of these proteins were investigated in ten cases of human glioma and two meningioma tissues and eight human glioma cell lines. In gelatin zymography, MMP-2 activities of glioblastomas were higher than astrocytomas. The activated form of MMP-2 was seen in five of six cases of glioblastomas, but not in astrocytomas. MMP-9 activity was detected in all cases of malignant astrocytomas but the reactivity of MMP-9 was weaker than that of MMP-2. MT1-MMP mRNA expression in glioblastomas was higher than that in astrocytomas. Five cases of glioblastomas with activated form of MMP-2 had MT1-MMP expressions. In vitro, human glioma cell lines with high expression of MT1-MMP also showed high MMP-2 activity. TIMP-1 transcripts were constitutively present in almost all glioma tissues and cell lines, whereas TIMP-2 mRNA were weak especially in malignant gliomas. Imbalance of TIMP-2/MMP-2 was observed using immunoprecipitation analysis in a glioma cell line. High expression of MMP-2 and MT1-MMP is possibly involved in invasiveness of malignant glioma.
Assuntos
Humanos , Animais , Northern Blotting/métodos , Encéfalo/patologia , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/enzimologia , Ativação Enzimática , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Regulação Enzimológica da Expressão Gênica , Glioma/patologia , Glioma/enzimologia , Metaloendopeptidases/genética , Papio , Inibidor Tecidual de Metaloproteinase-2/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Células Tumorais CultivadasRESUMO
To evaluate possible roles of matrix metalloproteinase (MMP)-1, -2, tissue inhibitor of metalloproteinase (TIMP)-1, -2 and membrane-type-1 matrix metalloproteinase (MT1-MMP) in invasion of human gliomas, expressions of these proteins were investigated in ten cases of human glioma and two meningioma tissues and eight human glioma cell lines. In gelatin zymography, MMP-2 activities of glioblastomas were higher than astrocytomas. The activated form of MMP-2 was seen in five of six cases of glioblastomas, but not in astrocytomas. MMP-9 activity was detected in all cases of malignant astrocytomas but the reactivity of MMP-9 was weaker than that of MMP-2. MT1-MMP mRNA expression in glioblastomas was higher than that in astrocytomas. Five cases of glioblastomas with activated form of MMP-2 had MT1-MMP expressions. In vitro, human glioma cell lines with high expression of MT1-MMP also showed high MMP-2 activity. TIMP-1 transcripts were constitutively present in almost all glioma tissues and cell lines, whereas TIMP-2 mRNA were weak especially in malignant gliomas. Imbalance of TIMP-2/MMP-2 was observed using immunoprecipitation analysis in a glioma cell line. High expression of MMP-2 and MT1-MMP is possibly involved in invasiveness of malignant glioma.
Assuntos
Humanos , Animais , Northern Blotting/métodos , Encéfalo/patologia , Neoplasias Encefálicas/patologia , Neoplasias Encefálicas/enzimologia , Ativação Enzimática , Metaloproteinase 2 da Matriz/metabolismo , Metaloproteinase 2 da Matriz/genética , Metaloproteinase 9 da Matriz/metabolismo , Regulação Enzimológica da Expressão Gênica , Glioma/patologia , Glioma/enzimologia , Metaloendopeptidases/genética , Papio , Inibidor Tecidual de Metaloproteinase-2/genética , Inibidor Tecidual de Metaloproteinase-1/genética , Células Tumorais CultivadasRESUMO
BACKGROUND: Matrix metalloproteinases (MMPs) have been implicated in the remodelling of extracellular matrix (ECM), including basement membrane. ECM remodelling is associated with pathological processes, including hepatic fibrosis, tumor invasion and metastasis. Tissue inhibitors of metalloproteinase (TIMP)-1 and TIMP-2 were known to inhibit MMP-9 and MMP-2, respectively. In the present study, we examined the expression of TIMP-1 and TIMP-2 in surgical specimen pairs of hepatocellular carcinoma and nontumoral liver and the correlation between their expression and clinicopathological characteristics. METHODS: The localization of both transcripts and protein of TIMP-1 and TIMP-2 was studied by using in situ hybridization and immunohistochemistry. RESULTS: TIMP-1 and TIMP-2 mRNA transcripts were found in tumor cells, hepatocyte, sinusoidal cells, endothelial cells and stromal cells. Signal intensity of TIMP-1 was stronger than that of TIMP-2. The results of immunohistochemical stainings were concordant with those obtained by in situ hybridization. Expression of TIMP-1 and TIMP-2 was observed in tumorous tissue, in nontumorous tissue and in the portions of the tumors adjacent to the capsules. However, a clear difference in TIMP-1 and TIMP-2 mRNA expression was not observed among the three tissue types. The intensity of TIMP-2 expression was generally weaker than that of TIMP-1, and the intensity of TIMP-1 and TIMP-2 mRNA expression did not correlate with variable clinicopathological characteristics. CONCLUSION: TIMPs was expressed in tumor cells and many cell types of the nontumoral liver. Further investigations for TIMPs' unknown functional role are needed.
Assuntos
Adulto , Idoso , Feminino , Humanos , Masculino , Carcinoma Hepatocelular/patologia , Carcinoma Hepatocelular/metabolismo , Imuno-Histoquímica , Neoplasias Hepáticas/patologia , Neoplasias Hepáticas/metabolismo , Pessoa de Meia-Idade , RNA Mensageiro/análise , Inibidor Tecidual de Metaloproteinase-2/fisiologia , Inibidor Tecidual de Metaloproteinase-2/genética , Inibidor Tecidual de Metaloproteinase-2/análise , Inibidor Tecidual de Metaloproteinase-1/fisiologia , Inibidor Tecidual de Metaloproteinase-1/genética , Inibidor Tecidual de Metaloproteinase-1/análiseRESUMO
Increased production of matrix metalloproteinases (MMPs) has been associated with increases in invasive and metastatic potential in many types of human carcinoma. Tissue inhibitors of metalloproteinase (TIMP)-1 inhibits most interstitial collagenases and MMP-9. TIMP-2 binds specifically and noncovalently to the pro-form of MMP-2 and inhibits its enzyme activity. In this study, we examined TIMP-1 and TIMP-2 expressions in relation to clinicopathological variables in colorectal carcinoma with in situ hybridization and immunohistochemistry. TIMP-1 and TIMP-2 expressions were localized overwhelmingly to pericancer stromal cells, while malignant and normal mucosal cells were weak or negative. Strong stromal TIMP-1 immunoreactivity correlated with Dukes' stage (p=0.022), status of lymph node metastasis (p=0.044) and poor survival (p= 0.005). The degree of immunohistochemical staining of TIMP-2 did not correlate with all clinicopathological variables. The correlation between enhanced TIMP-1 expression and advanced stage and poor survival suggest a growth promoting activity of TIMP-1 in colorectal carcinoma.