RESUMO
The incidence of prostate cancer (PCa) is increasing, and it is currently the second most frequent cause of death by cancer in men. Despite advancements in cancer therapies, new therapeutic approaches are still needed for treatment-refractory advanced metastatic PCa. Cross-species analysis presents a robust strategy for the discovery of new potential therapeutic targets. This strategy involves the integration of genomic data from genetically engineered mouse models (GEMMs) and human PCa datasets. Considering the role of antioxidant pathways in tumor initiation and progression, we searched oxidative stress-related genes for a potential therapeutic target for PCa. First, we analyzed RNA-sequencing data from Pb-Cre4; Ptenf/f mice and discovered an increase in sulfiredoxin (Srxn1) mRNA expression in high-grade prostatic intraepithelial neoplasia (PIN), well-differentiated adenocarcinoma (medium-stage tumors), and poor-differentiated adenocarcinoma (advanced-stage prostate tumors). The increase of SRXN1 protein expression was confirmed by immunohistochemistry in mouse prostate tumor paraffin samples. Analyses of human databases and prostate tissue microarrays demonstrated that SRXN1 is overexpressed in a subset of high-grade prostate tumors and correlates with aggressive PCa with worse prognosis and decreased survival. Analyses in vitro showed that SRXN1 expression is also higher in most PCa cell lines compared to normal cell lines. Furthermore, siRNA-mediated downregulation of SRXN1 led to decreased viability of PCa cells LNCaP. In conclusion, we identified the antioxidant enzyme SRXN1 as a potential therapeutic target for PCa. Our results suggest that the use of specific SRXN1 inhibitors may be an effective strategy for the adjuvant treatment of castration-resistant PCa with SRXN1 overexpression.
Assuntos
Terapia de Alvo Molecular , Oxirredutases atuantes sobre Doadores de Grupo Enxofre/metabolismo , Neoplasias da Próstata/patologia , Animais , Linhagem Celular Tumoral , Sobrevivência Celular , Progressão da Doença , Regulação Neoplásica da Expressão Gênica , Humanos , Masculino , Camundongos , Gradação de Tumores , Invasividade Neoplásica , Metástase Neoplásica , Estadiamento de Neoplasias , Estresse Oxidativo/genética , Oxirredutases atuantes sobre Doadores de Grupo Enxofre/genética , Prognóstico , Próstata/patologia , Neoplasias da Próstata/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Análise de SobrevidaRESUMO
Infravesical obstruction (IVO) secondary to benign prostatic hypertrophy can affect up to 50% of men over 50 years old and may cause serious and irreversible alterations throughout the urinary tract, especially in the bladder. Therapeutic approaches are currently limited. Amitriptyline has recently been described as an analgesic, anti-inflammatory and myorelaxant in some experimental models. The objective of this study was to investigate the effects of amitriptyline hydrochloride on the process of fibrosis in a bladder outlet obstruction model in rats. Male Wistar rats were subjected to IVO and studied at intervals of 1 and 14 days postprocedure. The rats were randomly divided into five groups: sham, IVO1-T, IVO1-NT, IVO14-T and IVO14-NT. Bladder tissue was processed for histopathology, immunohistochemistry and RT-PCR. The IVO14 groups presented bladder fibrosis, smooth muscle cell hypertrophy and bladder wall thickening. The IVO14-T group demonstrated a higher intensity of fibrosis, higher macrophage infiltration rate and higher gene expression of Transforming growth factor (TGF) Tgf-ß1. Thus this data shows that in this experimental mode amitriptyline had an amplifying effect on the process of fibrosis as a whole.
Assuntos
Amitriptilina/farmacologia , Obstrução do Colo da Bexiga Urinária/patologia , Bexiga Urinária/efeitos dos fármacos , Animais , Modelos Animais de Doenças , Fibrose , Masculino , Ratos , Ratos Wistar , Fator de Crescimento Transformador beta1/genética , Fator de Crescimento Transformador beta1/metabolismo , Bexiga Urinária/metabolismo , Bexiga Urinária/patologia , Obstrução do Colo da Bexiga Urinária/genética , Obstrução do Colo da Bexiga Urinária/metabolismoRESUMO
The purpose of this investigation was to analyze the immunoexpression of FasL, Fas, FADD, cleaved caspase 8, and cleaved caspase 3 in gastric cancer. Formalin-fixed and paraffin-embedded gastric adenocarcinoma tissues from 87 patients, including adjacent normal tissues, were included on tissue microarray by immunohistochemistry. The tumor and the adjacent normal tissues were positive for FasL in 66.7% and 90.6%, for Fas in 52.8% and 52.4%, for FADD in 67.4% and 82.3%, for cleaved caspase 8 in 27.9% and 37.7%, and for cleaved caspase 3 in 33.7% and 8.3%, respectively. FasL and the FADD from tumor were statistically different in relation to the histological type. Cleaved caspase 8 was statistically different in relation to clinical stage (p=0.031). The FADD from normal tissue was statistically different in relation to age (p=0.039), sex (p=0.055), clinical stage (p=0.019), and Fas was different in relation to tumor size (p=0.012). In the tumor, we observed a correlation between FasL and Fas, FasL and FADD, and FasL and cleaved caspase 3. In the adjacent normal tissue, a correlation was observed between FasL and Fas, FasL and FADD. There was no association of another marker with sex, age, clinical stage, and survival. Our results suggest that these proteins mediate the early extrinsic apoptotic pathway in gastric cancer and adjacent normal mucosa. FasL protein binds to Fas protein and subsequently binds to death receptor FADD signaling activation of the extrinsic apoptotic pathway. In this phase, there was inhibition of caspase 8 and, consequently, decreased apoptosis.
Assuntos
Adenocarcinoma/metabolismo , Adenocarcinoma/patologia , Apoptose/fisiologia , Transdução de Sinais/fisiologia , Neoplasias Gástricas/metabolismo , Neoplasias Gástricas/patologia , Adulto , Idoso , Idoso de 80 Anos ou mais , Caspase 3/análise , Caspase 8/análise , Proteína Ligante Fas/análise , Proteína de Domínio de Morte Associada a Fas/análise , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Análise Serial de Tecidos , Receptor fas/análiseRESUMO
This study was undertaken to investigate the immunoexpression of Bcl-2 family proteins (Bcl-2, Bcl-x, Bax, Bak and Bad) and to evaluate the correlation between the immunoexpression of these proteins and that of cleaved caspase 3, Ki-67 and p53. A tissue microarray (TMA) paraffin block was constructed using gastric carcinoma tissue (test group) and adjacent normal gastric mucosa (control group) from 87 patients who had not previously undergone radiotherapy or chemotherapy. Sections from the TMA block (4 µm) were subjected to immunohistochemistry for Bcl-2, Bcl-x, Bax, Bak, Bad, p53, Ki-67 and cleaved caspase-3. The slides were evaluated by the semi-quantitative method, and the scores obtained (intensity vs. percentage of staining) were correlated with one another and with the apoptotic index, cellular proliferation and data regarding patient survival. The studied proteins were present in the tumor tissue and in the normal gastric mucosa, but at different intensities and with differences in the number of positive cells. There was an association between tumor size and p53 expression, and intestinal type adenocarcinoma was positively correlated with the expression of Bax, Bad and Ki-67. The immunoexpression of Bcl-x, Bak, Bad, p53 and Ki-67 showed statistically significant differences between the tumor tissue and the adjacent normal gastric mucosa. There was an association between the expression of Bax, Bak and Bad in the normal gastric mucosa. No correlation between patient survival and the expression of these proteins was observed. Overexpression of the Bcl-x protein in the adenocarcinomas and the difference in Bcl-x expression between the test and the control group may be related to the anti-apoptotic effect of this protein. The reduced expression of Bak and Bad and the increased expression of p53 and Ki-67 in the adenocarcinomas demonstrate the imbalance between apoptosis and cellular proliferation, which results in uncontrolled tumor cell proliferation.