ABSTRACT
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.
Subject(s)
Humans , Carcinoma, Hepatocellular , Cell Movement/physiology , Liver Neoplasms , Matrix Metalloproteinase 9/genetics , Signal Transduction/physiology , Tissue Inhibitor of Metalloproteinase-1/genetics , Cell Line, Tumor/cytology , Cell Line, Tumor/physiology , Collagenases/genetics , Dipeptides/pharmacology , Matrix Metalloproteinase Inhibitors , Matrix Metalloproteinase 1/genetics , /genetics , /genetics , /genetics , Protease Inhibitors/pharmacology , Signal Transduction/drug effects , TransfectionABSTRACT
Gelatinase B or collagenase type IV is a 92 kDa protein. In case of over-expression of the gene, because of its collagenase activity, it can be involved in metastasis activity of few cancers e.g. bladder, colorectal and gastric carcinoma. Single nucleotide substitution base polymorphism of C to T at -1562 of promotor region can increase gene expression by decreasing transcription inhibitor proteins binding at T alleles. The aim of this case-control study is to investigate the role of this polymorphism in development and invasion of breast cancer in Isfahan women population. At this study 90 breast cancer patients with metastasis and 100 healthy controls were analyzed using polymerase chain reaction-restriction fragment length polymorphism assay [PCR-RFLP]. The mean of follow up period was 2 years. Patients were checked every 3-5 months Data analysis showed a close association between the presence of T allele and invasion of breast cancer [OR = 5.85, 95% CI: 2.64-12.93, p <0.0001]. According to our findings, the major role of this polymorphism is in cancer cell metastasis and invasion of these cells to adjacent tissues
Subject(s)
Humans , Female , Breast Neoplasms/analysis , Breast Neoplasms/genetics , Breast Neoplasms/complications , Breast Neoplasms/secondary , Case-Control Studies , Gelatinases/analysis , Gelatinases/genetics , Collagenases/analysis , Collagenases/genetics , Collagenases , Polymerase Chain Reaction , Polymorphism, Genetic/analysis , Polymorphism, Genetic/geneticsABSTRACT
The treatment of some mesenchymal malignancies has made significant gains over the past few decades with the development of effective systemic therapies. In contrast, the treatment of chondrosarcoma has been limited to surgical resection, with the most significant prognostic indicators being surgical margins and histologic grade. We have reported that MMP-1/TIMP-1 gene expression serves to prognosticate for tumor recurrence in this group of patients. This led to the hypothesis that collagenase activity facilitates cell egression from the cartilaginous matrix. In the current study we examine the specificity of collagenase gene expression in archival human chondrosarcoma samples using semi-quantitative PCR. Messenger RNA was affinity extracted and subject to reverse transcription. The subsequent cDNA was amplified using novel primers and quantitated by densitometry. Ratios of gene expression were constructed and compared to disease-free survival. The data demonstrate that the significance of the MMP-1/TIMP-1 ratio as a predictor of recurrence is confirmed with a larger number of patients. Neutrophil collagenase or MMP-8 was observed in only 5 of 29 samples. Collagenase-3 or MMP-13 was observed in all samples but the level did not correlate with disease-free survival. Since the collagenases have similar activity for fibrillar collagens and cleave the peptide in the same location, post-transcriptional regulatory mechanisms may account for the observed specificity. The determination of the MMP-1/TIMP-1 gene expression ratio not only serves to identify those patients at risk for recurrence but may also serve as a novel therapeutic avenue as an adjunct to surgical resection
Subject(s)
Humans , Bone Neoplasms/enzymology , Chondrosarcoma/enzymology , Collagenases/metabolism , Gene Expression Regulation, Enzymologic , Collagenases/analysis , Collagenases/genetics , Disease-Free Survival , DNA, Complementary/analysis , Polymerase Chain Reaction , Prognosis , Recurrence/prevention & control , RNA, Messenger/analysis , Substrate Specificity/genetics , Tissue Inhibitor of Metalloproteinase-1/analysisABSTRACT
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.
Subject(s)
Adult , Aged , Aged, 80 and over , Female , Humans , Male , Adenocarcinoma/pathology , Adenocarcinoma/mortality , Adenocarcinoma/enzymology , Antibodies , Collagenases/immunology , Collagenases/genetics , Collagenases/analysis , Colorectal Neoplasms/pathology , Colorectal Neoplasms/mortality , Colorectal Neoplasms/enzymology , DNA Probes , Matrix Metalloproteinase 2 , Matrix Metalloproteinase 9 , Gelatinases/immunology , Gelatinases/genetics , Gelatinases/analysis , Gene Expression Regulation, Enzymologic , Gene Expression Regulation, Neoplastic , In Situ Hybridization , Metalloendopeptidases/immunology , Metalloendopeptidases/genetics , Metalloendopeptidases/analysis , Middle Aged , Predictive Value of Tests , RNA, Messenger/analysis , Stromal Cells/pathology , Stromal Cells/enzymology , Survival Analysis , Tissue Inhibitor of Metalloproteinase-2/immunology , Tissue Inhibitor of Metalloproteinase-2/genetics , Tissue Inhibitor of Metalloproteinase-2/analysis , Tissue Inhibitor of Metalloproteinase-1/immunology , Tissue Inhibitor of Metalloproteinase-1/genetics , Tissue Inhibitor of Metalloproteinase-1/analysisABSTRACT
The equilibrium between deposition and degradation of extracellular matrix(ECM) is essential to normal tissue development and repair of wound or inflammatory responses. It has recently become apparent that several cytokines and growth factors are capable of modulating fibroblast proliferation and biosynthetic activity. To understand the role of these factors in connective tissue regulation, we examined the effect of interferon-gamma (IFN-gamma) on stromelysin-1 gene expression in cultured human dermal fibroblasts. The steady-state levels of stromelysin-1 mRNA were increased in IFN-gamma treated cultured dermal fibroblasts. In the CAT assay, the stromelysin-1 promoter activity was increased 2.8-fold compared with untreated control. Therefore IFN-gamma stimulates the stromelysin-1 promoter activity, resulting in transcriptional enhancement of gene expression. Transforming growth factor-beta (TGF-beta) showed the antagonistic action to the effects of IFN-gamma in cultured dermal fibroblasts. Furthermore, gel mobility shift assays demonstrated enhanced AP-1 binding activities in nuclear extracts from cells incubated with IFN-gamma. These data suggest that IFN-gamma is an up-regulator and TGF-beta is a down regulator on the stromelysin-1 gene expression, respectively, and the AP-1 binding site may be necessary for gene response.