Navegando por Palavras-chave "Endostatin"
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- ItemAcesso aberto (Open Access)Collagen XVIII/endostatin expression in experimental endotoxemic acute renal failure(Associação Brasileira de Divulgação Científica, 2009-12-01) Cichy, Milene Cristina [UNIFESP]; Rocha, Flavia Gomes de Goes [UNIFESP]; Tristão, Vivian Regina [UNIFESP]; Pessoa, Edson de Andrade [UNIFESP]; Cenedeze, Marcos Antonio [UNIFESP]; Nürmberg Junior, R.; Schor, Nestor [UNIFESP]; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Instituto de Pesquisas Energéticas e Nucleares Centro de Biotecnologia; Faculdades Metropolitanas Unidas Faculdade de Medicina VeterináriaAcute renal failure (ARF) is a frequent complication of Gram-negative sepsis, with a high risk of mortality. Lipopolysaccharide (LPS)-induced ARF is associated with hemodynamic changes that are strongly influenced by the overproduction of nitric oxide (NO) through the cytokine-mediated up-regulation of inducible NO synthase. LPS-induced reductions in systemic vascular resistance paradoxically culminate in renal vasoconstriction. Collagen XVIII is an important component of the extracellular matrix expressed in basement membranes. Its degradation by matrix metalloproteases, cathepsins and elastases results in the formation of endostatin, claimed to have antiangiogenic activity and to be a prominent vasorelaxing agent. We evaluated the expression of endostatin/collagen XVIII in an endotoxemic ARF model. ARF was induced in C57BL/6 mice by intraperitoneal injection of LPS (10 mg/kg) followed by sacrifice 4 and 12 h later. Kidney tissue was the source of RNA and protein and the subject of histological analysis. As early as 4 h after LPS administration, blood urea, creatinine and NO levels were significantly increased compared to control. Endostatin/collagen XVIII mRNA levels were 0.71 times lower than sham-inoculated mice 4 h after LPS inoculation, returning to normal levels 12 h after LPS inoculation. Immunohistological examination revealed that acute injury caused by LPS leads to an increase of endostatin basement membrane staining in association with the decrease of CD31 endothelial basement membrane staining. These results indicate that in the early phase of endotoxemic ARF the endostatin levels were not regulated by gene expression, but by the metabolism of collagen XVIII.
- ItemSomente MetadadadosEndostatin gene therapy enhances the efficacy of IL-2 in suppressing metastatic renal cell carcinoma in mice(Springer, 2010-09-01) Goes Rocha, Flavia Gomes de [UNIFESP]; Barbosa Chaves, Karen Cristina [UNIFESP]; Chammas, Roger; Schatzmann Peron, Jean Pierre; Rizzo, Luiz Vicente; Schor, Nestor [UNIFESP]; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP); Albert Einstein Jewish Inst Educ & Res; IPEN CNENWe investigated whether the administration of IL-2 combined with endostatin gene therapy was able to produce additive or even synergistic immunomodulatory activity in a mouse model of metastatic renal carcinoma. Renca cells were injected into the tail vein of BALB/c mice. After 24 h, the animals were randomly divided into four groups (5 mice/group). One group of mice was the control, the second group received treatment with 100,000 UI of Recombinant IL-2 (Proleukin, Chiron) twice a day, 1 day per week during 2 weeks (IL-2), the third group received treatment with a subcutaneous inoculation of 3.6 x 10(6) endostatin-producing cells, and the fourth group received both therapies (IL-2 + ES). Mice were treated for 2 weeks. in the survival studies, 10 mice/group daily, mice were monitored daily until they died. the presence of metastases led to a twofold increase in endostatin levels. Subcutaneous inoculation of NIH/3T3-LendSN cells resulted in a 2.75 and 2.78-fold increase in endostatin levels in the ES and IL-2 + ES group, respectively. At the end of the study, there was a significant decrease in lung wet weight, lung nodules area, and microvascular area (MVA) in all treated groups compared with the control group (P < 0.001). the significant difference in lung wet weight and lung nodules area between groups IL-2 and IL-2 + ES revealed a synergistic antitumor effect of the combined treatment (P < 0.05). the IL-2 + ES therapy Kaplan-Meier survival curves showed that the probability of survival was significantly higher for mice treated with the combined therapy (log-rank test, P = 0.0028). Conjugated therapy caused an increase in the infiltration of CD4, CD8 and CD49b lymphocytes. An increase in the amount of CD8 cells (P < 0.01) was observed when animals received both ES and IL-2, suggesting an additive effect of ES over IL-2 treatment. A synergistic effect of ES on the infiltration of CD4 (P < 0.001) and CD49b cells (P < 0.01) was also observed over the effect of IL-2. Here, we show that ES led to an increase in CD4 T helper cells as well as cytotoxic lymphocytes, such as NK cells and CD8 cells, within tumors of IL-2 treated mice. This means that ES plays a role in supporting the actions of T cells.
- ItemAcesso aberto (Open Access)Endostatin gene therapy inhibits intratumoral macrophage M2 polarization(Elsevier France-Editions Scientifiques Medicales Elsevier, 2016) Foguer, Karen [UNIFESP]; Braga, Marina de Souza [UNIFESP]; Schatzmann Peron, Jean Pierre; Bortoluci, Karina Ramalho [UNIFESP]; Bellini, Maria Helena [UNIFESP]Background: Renal cell carcinoma (RCC) is a highly vascularized cancer resistant to chemotherapy and radiotherapy. RCC is frequently infiltrated with immune cells, with macrophages being the most abundant cell type. Alternatively activated M2 macrophages are known to contribute to tumor progression. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. In this study, we investigated the impact of ES gene therapy on the polarization of tumor-associated macrophages (TAMs) in lung metastases from tumor-bearing mice. Methods: BALB/c mice divided into three groups: Normal, Control and ES-treated. Tumor-bearing mice were treated with ES-transduced cells or control cells over ten days. At the end of the study, plasma was collected, and pulmonary macrophages were isolated and used for FACS or RT-PCR. ELISA tests were used to analyze plasma and cell culture supernatant cytokines. Results: ES treatment significantly reduced the levels of anti-inflammatory and pro-angiogenic cytokines, including IL4, IL-10, IL-13 and VEGF. Gene expression of M2 markers, such as IL-10, Arg-1, VEGF and YM-1, declined significantly. Flow cytometry showed a reduction in the number of M2 F4/80 + CD36 + CD206 + CD209+ macrophages and in IL-10 secretion by these cells. Reduced levels of IL-10 were also found in the culture supernatants of the ES-treated group. Conclusions: Our research corroborates previous observations that ES has an important anti-tumoral role. However, aside from promoting interferon-g secretion and an effective T cell response, we show here that this switch is extended to TAMs, complicating the maintenance of pro-tumorigenic M2 macrophages and thus favoring tumor elimination. (C) 2016 Elsevier Masson SAS. All rights reserved.
- ItemSomente MetadadadosEndostatin gene therapy stimulates upregulation of ICAM-1 and VCAM-1 in a metastatic renal cell carcinoma model(Nature Publishing Group, 2012-08-01) Chaves, Karen Cristina Barbosa [UNIFESP]; Peron, Jean Pierre Schatzmann; Chammas, Roger; Turaca, Lauro Thiago [UNIFESP]; Pesquero, João Bosco [UNIFESP]; Braga, Marina de Souza [UNIFESP]; Foguer, Karen [UNIFESP]; Schor, Nestor [UNIFESP]; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP)One of the greatest challenges in urological oncology is renal cell carcinoma (RCC), which is the third leading cause of death in genitourinary cancers. RCCs are highly vascularized and respond positively to antiangiogenic therapy. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. in this study, we examined the potential of ES-based antiangiogenic therapy to activate tumor-associated endothelial cells in metastatic RCC (mRCC). Balb/c-bearing Renca cells were treated with NIH/3T3-LendSN or, as a control, with NIH/3T3-LXSN cells. the T-cell subsets and lymphocyte populations of tumors, mediastinal lymph nodes and the spleen were assessed by flow cytometry. the expression of intercellular adhesion molecule-1 (ICAM-1) and vascular cell adhesion molecule-1 (VCAM-1) was assessed by real-time PCR, flow cytometry and immunohistochemistry analysis. ES gene therapy led to an increase in the percentage of infiltrating CD4-interferon (IFN)-gamma cells (P<0.05), CD8-IFN-gamma cells (P<0.01) and CD49b-tumor necrosis factor-alpha cells (P<0.01). in addition, ES therapy caused an increase at the mRNA level of ICAM-1 (1.4-fold; P<0.01) and VCAM-1 (1.5-fold) (control vs treated group; P<0.001). Through flow cytometry, we found a significant increase in the CD34/ICAM-1 cells (8.1-fold; P<0.001) and CD34/VCAM-1 cells (1.6-fold; P<0.05). ES gene therapy induced a significant increase in both T CD4 and CD8 cells in the lymph nodes and the spleen, suggesting that ES therapy may facilitate cell survival or clonal expansion. CD49b cells were also present in increased quantities in all of these organs. in this study, we demonstrate an antitumor inflammatory effect of ES in an mRCC model, and this effect is mediated by an increase in ICAM-1 and VCAM-1 expression in tumor-associated endothelial cells.
- ItemSomente MetadadadosEndostatin neoadjuvant gene therapy extends survival in an orthotopic metastatic mouse model of renal cell carcinoma(Elsevier B.V., 2012-06-01) Braga, Marina de Souza [UNIFESP]; Chaves, Karen Barbosa [UNIFESP]; Chammas, Roger [UNIFESP]; Schor, Nestor [UNIFESP]; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Dept BiotechnolDespite recent advances in targeted therapy, renal cell carcinoma (RCC) remains one of the most lethal urologic malignancies. Approximately 30% of patients with localised RCC will develop metastases after curative surgery. Presurgical therapy has been explored for treatment of localised RCC. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. in this study, we examined the potential use of an antiangiogenic agent as a neoadjuvant therapy in an orthotopic metastatic mouse model of RCC. BALB/c mice bearing Renca cells were treated before nephrectomy with NIH/3T3-LendSN cells. At the end of the experiment, ES serum levels were measured. Primary and metastatic tumour area and microvascular area were determined. in the survival studies, mice were monitored daily until they died. ES serum levels in treated mice were higher in the control group (P < 0.05). the median primary tumour area and the mean microvascular area were significantly lower in the ES-treated group compared to control group (P < 0.05). the proliferation of Renca cells in the ES-treated group was significantly reduced compared with the control group (P < 0.01). ES therapy led to a significant reduction in the number of pulmonary metastatic nodules compared with the control group (P < 0.01). Kaplan-Meier survival curves showed that the probability of survival was significantly higher in mice receiving ES therapy (P = 0.0243, Log-Rank test). Our results indicated that neoadjuvant ES gene therapy has the potential to decrease tumour burden, extend survival, and may have clinical benefit in the management of RCC. (C) 2011 Elsevier Masson SAS. All rights reserved.
- ItemSomente MetadadadosEndostatin, an antiangiogenic protein, is expressed in the unilateral ureteral obstruction mice model(Wichtig Editore, 2008-09-01) Maciel, Thiago T. [UNIFESP]; Coutinho, Enia L. [UNIFESP]; Soares, Debora [UNIFESP]; Achar, Eduardo [UNIFESP]; Schor, Nestor [UNIFESP]; Bellini, Maria H. [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); IPEN CNEN SPBackground: Extracellular matrix accumulation, epithelial-to-mesenchymal transition, tubular atrophy and loss of peritubular capillary network are hallmarks of tubulointerstitial injury in progressive renal diseases. In this study, we analyzed endostatin expression in kidneys subjected to unilateral ureteral obstruction (UUO).Methods: Collagen XVIII mRNA expression was evaluated by real-time polymerase chain reaction (PCR). Endostatin and CD31 protein levels were analyzed by Western blot and immunohistochemistry. In vitro quantification of collagen XVIII and fibrosis-related genes in HK2 cells was performed by real-time PCR.Results: UUO significantly increased collagen XVIII mRNA expression and released a 30-kDa endostatin fragment. Immunohistochemistry revealed endostatin expression increased in injured tissue, mainly on tubular cells. Of interest, expression of CD31 was significantly reduced by UUO. Endostatin administration in vitro did not modify the expression of genes related to fibrosis development. However, in vitro TGF-b1 administration induced expression of collagen XVIII/endostatin mRNA in human tubular cells.Conclusion: Endostatin is expressed during the progression of renal fibrosis in vitro and in vivo, suggesting a role for endostatin in development of tubulointerstitial injury.
- ItemSomente MetadadadosErythrocyte Protoporphyrin Fluorescence as a Biomarker for Monitoring Antiangiogenic Cancer Therapy(Springer, 2010-11-01) Goes Rocha, Flavia Gomes de [UNIFESP]; Barbosa Chaves, Karen Cristina [UNIFESP]; Gomes, Cinthia Zanini; Campanharo, Camila Barricheli [UNIFESP]; Courrol, Lilia Coronato [UNIFESP]; Schor, Nestor [UNIFESP]; Bellini, Maria Helena [UNIFESP]; IPEN CNEN SP; Universidade Federal de São Paulo (UNIFESP)Renal cell carcinoma (RCC) remains one of the greatest challenges of urological oncology and is the third leading cause of death in genitourinary cancers. RCCs are highly vascularized and are amenable to antiangiogenic therapy. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. in this study, we examined the potential of erythrocyte PpIX fluorescence spectroscopy for monitoring the efficacy of antiangiogenic therapy in metastatic renal cell carcinoma (mRCC), using an orthotopic metastatic mouse model. Balb/C-bearing Renca cells were treated with NIH/3T3-LendSN cells. Lung weight, nodule area, microvascular area (MVA), and erythrocyte PpIX fluorescence were evaluated. Emission spectra were obtained by exciting the samples at 405 nm. There was a significant decrease in lung wet weight, lung nodule area and MVA in the treated group compared to the control group (P < 0.001). Significant differences in autofluorescence shape were observed in the 620-650 nm spectral region. the most intense fluorescence peak was observed at similar to 632 nm. the autofluorescence of the control samples was about 53% higher than that of normal blood (P < 0.05). in the group treated with ES, the autofluorescence was about 54% lower than in the control group (P < 0.05). Fluorescence intensity was positively correlated with the nodule area (R (2) = 0.8859; P < 0.001) and MVA (R (2) = 0.9431; P < 0.001) in the ES-treated group. These results demonstrate that the spectroscopic analysis method allows a selective detection of tumor masses. This preliminary study suggests that PpIX fluorescence may be useful as a biomarker for antiangiogenic cancer therapy.
- ItemAcesso aberto (Open Access)Estudo da modulação das células mielóides supressoras no adenocarcinoma metastático após terapia antiangiogênica(Universidade Federal de São Paulo (UNIFESP), 2016-08-31) Chaves, Karen Cristina Barbosa [UNIFESP]; Bellini, Maria Helena [UNIFESP]; http://lattes.cnpq.br/4219112551635779; http://lattes.cnpq.br/1190409748118272; Universidade Federal de São Paulo (UNIFESP)Renal cell carcinoma (RCC), the third leading cause of death in genitourinary cancers. RCCs are highly vascularized and resistant to radiotherapy and chemotherapy. Antiangiogenic drugs are promising and widely used in clinical, on the other hand, mechanism of evasive resistance has been seen to treatment with anti-VEGF antibody. Recent reports suggest that treatment resistance of patients with cancer is related to the presence of myeloid-derived suppressor cells. Endostatin (ES) is a fragment of collagen XVIII that mediates antiangiogenic activity, however, its mechanisms of action are unclear. In this study, we tracked Gr-1+ cells in different organs, evaluated the role of CD11b+Gr-1+ cells and their subsets during tumoral progression and examined the therapeutic potential of ES in modulating CD11b+Gr-1+ cells and their subsets as well as their immunosuppressive activities in lung metastasis. Healthy Balb/c mice were used to detect Gr-1+ cells. CCRm-bearing mice were nephrectomized on day 7 and collected metastatic lung, spleen and bone marrow after 3, 7 and 10 days. Quantification of CD11b+Gr-1+ cells and their monocytic and granulocytic subsets was performed by flow cytometry and microscopic analysis were viewed in HE staining. CCRm-bearing mice were treated with NIH/3T3-LendSN-clone 5 or with NIH/3T3-LXSN cells as a control. ES and G-CSF levels were measured by ELISA and Milliplex, respectively. Quantification of CD11b+Gr-1+ cells and their subsets was performed by flow cytometry. Gr-1+ cells magnetically separated were evaluated in vitro the nitrite levels in supernatant and arginase activity in cells by colorimetric assays. ROS production was measured in CD11b+Gr-1+ cells using the DCFDA marker by flow cytometry. Our data showed the presence of CD11b+Gr-1+ cells in the bone marrow as well as in the kidney, lung and spleen, confirming the monocytic and granulocytic morphologies In metastatic progression, saw the expansion of CD11b+Gr-1+ cells and, preferably, the granulocytic subtype in the bone marrow. Our data demonstrate the progressive accumulation of splenic CD11b+Gr-1+ cells and the opposite was seen in metastatic lungs.Gene therapy with ES reduced the number of pulmonary metastatic lesions, the number of CD11b+Gr-1+ cells and the number of granulocytic cells. G-CSF levels were also reduced and ROS production by granulocytic cells was affected after the treatment with ES. Here, we demonstrate that the metastatic progression is inversely proportional to the number of CD11b+Gr-1+ cells present in the tumor microenvironment, showing that the granulocytic subtype is involved in advanced metastasis. Treatment with ES induced relevant antitumor immune response abrogating the reduction of ROS-producing myeloid-derived suppressor cells in the metastatic local.
- ItemSomente MetadadadosAn extracellular proteasome releases endostatin from human collagen XVIII(Springer, 2017) Reiss-Pistilli, Maria L. V.; Schuppan, Detlef; Barroso, Madalena M. S.; Assuncao-Miranda, Iranaia; Farias, Shirley [UNIFESP]; Lery, Leticia; Bauer, Michael; Juliano, Luiz [UNIFESP]; Juliano, Maria A. [UNIFESP]; Coelho-Sampaio, TatianaEndostatin is a potent anti-angiogenic and anti-tumor protein capable of regressing tumors without inducing acquired resistance. Since it is a fragment of the parental molecule, collagen XVIII, its endogenous production depends on the activity of a specific proteolytic enzyme. While such an enzyme has been described in mice, a human counterpart has not been identified so far. Here, we searched for this enzyme by using a fluorescence resonance energy transfer peptide containing the cleavage site of human collagen XVIII. We found that the cleavage activity was present in various murine and human tumor cells but not in untransformed cells. It was ascribed to a large protein complex identified as an extracellular form of proteasome 20S. Since circulating proteasome 20S has recently emerged as an important marker of tumor progression, the possibility of proteasomes controlling the production of angiostatic endostatin may inspire the development of new anticancer therapies.
- ItemSomente MetadadadosFibronectin expression is decreased in metastatic renal cell carcinoma following endostatin gene therapy(Elsevier B.V., 2012-09-01) Barbosa Chaves, Karen Cristina [UNIFESP]; Turaca, Lauro Thiago [UNIFESP]; Pesquero, Joao Bosco [UNIFESP]; Mennecier, Gregory; Zaidan Dagli, Maria Lucia; Chammas, Roger; Schor, Nestor [UNIFESP]; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP)Tumor cells induce the disruption of homeostasis between cellular and extracellular compartments to favor tumor progression. the expression of fibronectin (FN), a matrix glycoprotein, is increased in several carcinoma cell types, including renal cell carcinoma (RCC). RCC are highly vascularized tumors and are often amenable to antiangiogenic therapy. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. in this study, we examined the modulation of FN gene expression by ES gene therapy in a murine metastatic renal cell carcinoma (mRCC) model. Balb/C mice bearing Renca cells were treated with NIH/3T3-LXSN cells or NIH/3T3-LendSN cells. At the end of the experiment, the ES serum levels were measured, and the FN gene expression was assessed using real-time PCR. the tissue FN was evaluated by western blotting and by immunofluorescence analysis. the ES serum levels in treated mice were higher than those in the control group (P < 0.05). ES treatment led to significant decreases at the FN mRNA (P < 0.001) and protein levels (P < 0.01). Here, we demonstrate the ES antitumor effect that is mediated by down-regulation of FN expression in mRCC. (C) 2012 Elsevier Masson SAS. All rights reserved.
- ItemSomente MetadadadosImmobilized Kidney 28-kDa Endostatin- Related (KES28kDa) Fragment Promotes Endothelial Cell Survival(Karger, 2010-01-01) Bellini, Maria Helena [UNIFESP]; Malpighi, Thiago Franca; Calvo, Fernanda Bernardes; Miranda, Adriana Regina; Spencer, Patrick Jack; Cichy, Milena Cristina [UNIFESP]; Simons, Simone Michaela; Chudzinski Tavassi, Ana Marisa; Santos, Marinilce Fagundes dos; Rodrigues, Consuelo Junqueira; Schor, Nestor [UNIFESP]; CNEN SP; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP)Background/Objective: Renal ischemia-hypoxia is a leading cause of acute kidney injury (AKI). Ischemia causes extracellular matrix breakdown of the tubular basement membrane. Endostatin (ES) is the C-terminal fragment of collagen XVIII generated by proteolytic cleavage. Recent studies have demonstrated that ES expression is upregulated in ischemic kidneys. the present study aimed to characterize ES from ischemic kidneys. Methods: Ischemic renal failure was induced via 45 min of occlusion of the left renal artery and vein. After the ischemic period, blood was collected. Kidneys were harvested and used for immunohistochemical testing and protein extraction. Three-step purification was used. Soluble and immobilized purified ES were tested in cell viability and adhesion assays. Results: the soluble KES28kDa inhibited endothelial cell proliferation: 25 versus 12.5 mu g (p < 0.05); 12.5 versus 3.15 mu g (p < 0.05). Immobilization of KES28kDa supports endothelial cell survival over the control p = 0.021). Human umbilical vein endothelial cells plated on immobilized KES28kDa showed an increase in membrane ruffles and stress fibers. Conclusion: These data demonstrate the local synthesis of a 28-kDa ES-related fragment following AKI and suggest its role in endothelium survival. Copyright (C) 2010 S. Karger AG, Basel
- ItemSomente MetadadadosInvolvement of the NF-kappa B/p50/Bcl-3 complex in response to antiangiogenic therapy in a mouse model of metastatic renal cell carcinoma(Elsevier B.V., 2014-09-01) Braga, Marina de Souza [UNIFESP]; Silva Paiva, Katiucia Batista da; Foguer, Karen [UNIFESP]; Barbosa Chaves, Karen Cristina [UNIFESP]; Lima, Larissa de Sa; Scavone, Cristoforo; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP); IPEN CNENRenal cell carcinoma (RCC) represents approximately 2-3% of human malignancies. Nuclear transcription factor kappa B (NF-kappa B) is composed of a family of transcription factors that have been associated with the development and progression of RCC. Endostatin (ES) is a fragment of collagen XVIII that possesses antiangiogenic activity. in this study, we evaluated the expression of NF-kappa B in metastatic tumor cells from animals treated with ES. Balb/c-bearing Renca-EGFP cells were treated with NIH/3T3-LendSN or NIH/3T3-LXSN cells as a control. At the end of the in vivo experiment, plasma Renca-EGFP-sorted cells and tissue lung samples were collected. A real-time PCR array for NF-kappa B target genes revealed that ES therapy led to down regulation of Bcl-3 (P < 0.031), NF-kappa B1 (P < 0.001) and c-Rel (P < 0.004) in the ES-treated group. Using an electrophoretic mobility shift assay (EMSA), we observed a reduction in NF-kappa B binding activity in ES-treated Renca-EGP cells. Furthermore, a supershift assay showed a clear shift of the NF-kappa B DNA band in samples incubated with a p50 antibody. By immunohistochemistry analysis, ES treatment resulted in a significant reduction in expression of p50. (ES vs. control P < 0.05). the immunoprecipitation experiments confirmed the presence of a p50/Bcl-3 complex in nuclear extracts from cells of metastatic lung tissues. Our findings indicate that p50 and Bcl-3 plays a regulatory role in gene transcription in RCC. (C) 2014 Elsevier Masson SAS. All rights reserved.
- ItemSomente MetadadadosVascular endothelial growth factor as a biomarker for endostatin gene therapy(Elsevier B.V., 2013-07-01) Braga, Marina Souza [UNIFESP]; Turaca, Thiago Lauro [UNIFESP]; Foguer, Karen [UNIFESP]; Barbosa Chaves, Karen Cristina [UNIFESP]; Pesquero, Joao Bosco [UNIFESP]; Chammas, Roger; Schor, Nestor [UNIFESP]; Bellini, Maria Helena [UNIFESP]; Universidade Federal de São Paulo (UNIFESP); Universidade de São Paulo (USP); IPEN CNENRenal cell carcinoma (RCC) is characterized by high vascular endothelial growth factor (VEGF) production and, consequently, excessive angiogenesis. Several strategies have been developed to target angiogenesis as a method for treating metastatic RCC (mRCC). Endostatin (ES) is a C-terminal fragment of collagen XVIII that has antiangiogenic activity. the aim of this study was to investigate the predictive value of circulating VEGF-A in a murine model of mRCC after ES gene therapy. ES therapy did not affect the levels of collagen XVIII/ES or ES in the tissue. the circulating level of ES was increased in the control and ES-treated groups (normal vs. control, P < 0.05 and ES-treated vs. control, P < 0.001), and the intratumoral vessels were significantly decreased (ES-treated vs. control, P < 0.05). ES therapy decreased the VEGF mRNA levels. the tissue and circulating levels of VEGF in the control group were significantly higher than normal (P < 0.01 and P < 0.05, respectively). Treatment with ES significantly reduced the VEGF concentrations in both compartments (P < 0.001 for tissue and P < 0.05 for plasma). Our findings indicate that in addition to the directly targeted tumor vessels, ES exerts its antitumor effect by down-regulating VEGF gene expression in renal tumor cells. Additionally, our findings point to the predictive value of VEGF for ES therapy. (C) 2013 Elsevier Masson SAS. All rights reserved.