RESUMO
Urokinase-type plasminogen activator (uPA) and its receptor (uPAR) act in the proteolysis of basement membrane and extracellular matrix structures, facilitating tumor invasion. The purpose of this study was to evaluate the relationship between these proteins and clinicopathological parameters in squamous cell carcinoma of the oral tongue (SCCOT). Sixty cases of SCCOT were submitted to immunohistochemistry and analyzed semiquantitatively at the invasion front and in the tumor core. The results were associated with lymph node metastasis, clinical stage, locoregional recurrence, clinical outcome and histological grade of malignancy. A higher expression of uPA was observed in cases of tumors of high-grade versus low-grade malignancy (p = 0.010). Moreover, the cases with the worst pattern of invasion presented an overexpression of uPA (p = 0.011). The presence of locoregional recurrence was associated with uPAR (p = 0.039), and the expression of both biomarkers was much higher at the invasion front than in the tumor core (p < 0.001). The results suggest uPA and uPAR are involved in the progression and aggressiveness of SCCOT, mainly at the tumor-host interface.
Assuntos
Carcinoma de Células Escamosas/química , Receptores de Ativador de Plasminogênio Tipo Uroquinase/análise , Neoplasias da Língua/química , Ativador de Plasminogênio Tipo Uroquinase/análise , Biomarcadores Tumorais/análise , Carcinoma de Células Escamosas/patologia , Feminino , Humanos , Imuno-Histoquímica , Masculino , Pessoa de Meia-Idade , Gradação de Tumores , Invasividade Neoplásica , Recidiva Local de Neoplasia/química , Recidiva Local de Neoplasia/patologia , Estadiamento de Neoplasias , Valores de Referência , Fatores de Risco , Estatísticas não Paramétricas , Neoplasias da Língua/patologiaRESUMO
Abstract Urokinase-type plasminogen activator (uPA) and its receptor (uPAR) act in the proteolysis of basement membrane and extracellular matrix structures, facilitating tumor invasion. The purpose of this study was to evaluate the relationship between these proteins and clinicopathological parameters in squamous cell carcinoma of the oral tongue (SCCOT). Sixty cases of SCCOT were submitted to immunohistochemistry and analyzed semiquantitatively at the invasion front and in the tumor core. The results were associated with lymph node metastasis, clinical stage, locoregional recurrence, clinical outcome and histological grade of malignancy. A higher expression of uPA was observed in cases of tumors of high-grade versus low-grade malignancy (p = 0.010). Moreover, the cases with the worst pattern of invasion presented an overexpression of uPA (p = 0.011). The presence of locoregional recurrence was associated with uPAR (p = 0.039), and the expression of both biomarkers was much higher at the invasion front than in the tumor core (p < 0.001). The results suggest uPA and uPAR are involved in the progression and aggressiveness of SCCOT, mainly at the tumor-host interface.
Assuntos
Humanos , Masculino , Feminino , Neoplasias da Língua/química , Carcinoma de Células Escamosas/química , Ativador de Plasminogênio Tipo Uroquinase/análise , Receptores de Ativador de Plasminogênio Tipo Uroquinase/análise , Valores de Referência , Neoplasias da Língua/patologia , Imuno-Histoquímica , Carcinoma de Células Escamosas/patologia , Biomarcadores Tumorais/análise , Fatores de Risco , Estatísticas não Paramétricas , Gradação de Tumores , Pessoa de Meia-Idade , Invasividade Neoplásica , Recidiva Local de Neoplasia/patologia , Recidiva Local de Neoplasia/química , Estadiamento de NeoplasiasRESUMO
Prostatic lesions such as prostatic intraepithelial neoplasia (PIN) and proliferative inflammatory atrophy (PIA) are studied in human and canine species due to their malignance potential. The plasminogen activator (PA) system has been suggested to play a central role in cell adhesion, angiogenesis, inflammation, and tumor invasion. The urokinase-type plasminogen activator receptor (uPAR) is a component of the PA, with a range of expression in tumor and stromal cells. In this study, uPAR expression in both canine normal prostates and with proliferative disorders (benign prostatic hyperplasia-BPH, proliferative inflammatory atrophy-PIA, prostatic intraepithelial neoplasia-PIN, and carcinoma-PC) was evaluated by immunohistochemistry in a tissue microarray (TMA) slide to establish the role of this enzyme in extracellular matrix (ECM) remodeling and in the processes of tissue invasion. A total of 298 cores and 355 diagnoses were obtained, with 36 (10.1%) normal prostates, 46 (13.0%) with BPH, 128 (36.1%) with PIA, 74 (20.8%) with PIN and 71 (20.0%) with PC. There is variation in the expression of uPAR in canine prostate according to the lesion, with lower expression in normal tissue and with BPH, and higher expression in tissue with PIA, PIN and PC. The high expression of uPAR in inflammatory and neoplastic microenvironment indicates increased proteolytic activity in canine prostates with PIA, PIN, and PC.
Lesões prostáticas como a neoplasia intraepitelial prostática (PIN) e a atrofia inflamatória proliferativa (PIA) são estudadas na espécie humana e canina devido ao seu potencial de malignidade. O sistema de ativador de plasminogênio (PA) tem sido sugerido como um importante mecanismo na adesão celular, angiogênese, inflamação e invasão tumoral. O receptor ativador de plasminogênio do tipo uroquinase (uPAR) é um componente do PA, expresso em células tumorais e estromais. Avaliou-se, por imunoistoquímica em lâmina de microarranjo tecidual (TMA), a expressão de uPAR no tecido prostático canino normal e com desordens proliferativas (hiperplasia prostática benigna-HPB, atrofia inflamatória proliferativa-PIA, neoplasia epitelial prostática-PIN e carcinoma-CP), com o objetivo de verificar o papel desta enzima na remodelação de matriz extracelular (ECM) e no processo de invasão tecidual. Foram obtidos 298 cores e 355 diagnósticos, sendo 36 (10,1%) próstatas normais, 46 (13,0%) com HPB, 128 (36,1%) com PIA, 74 (20,8%) com PIN e 71 (20,0%) com CP. Há variação na expressão de uPAR na próstata canina de acordo com a lesão, com menor expressão nas glândulas normais e com HPB, e maior naquelas com lesões displásicas e (PIA e PIN) e neoplásicas (CP). A superexpressão de uPAR nos microambientes inflamatório e neoplásico indica aumento da atividade proteolítica em próstatas caninas com PIA, PIN e CP.
Assuntos
Masculino , Animais , Cães , Ativador de Plasminogênio Tipo Uroquinase/análise , Doenças do Cão , Neoplasia Prostática Intraepitelial/enzimologia , Neoplasia Prostática Intraepitelial/veterináriaRESUMO
Prostatic lesions such as prostatic intraepithelial neoplasia (PIN) and proliferative inflammatory atrophy (PIA) are studied in human and canine species due to their malignance potential. The plasminogen activator (PA) system has been suggested to play a central role in cell adhesion, angiogenesis, inflammation, and tumor invasion. The urokinase-type plasminogen activator receptor (uPAR) is a component of the PA, with a range of expression in tumor and stromal cells. In this study, uPAR expression in both canine normal prostates and with proliferative disorders (benign prostatic hyperplasia-BPH, proliferative inflammatory atrophy-PIA, prostatic intraepithelial neoplasia-PIN, and carcinoma-PC) was evaluated by immunohistochemistry in a tissue microarray (TMA) slide to establish the role of this enzyme in extracellular matrix (ECM) remodeling and in the processes of tissue invasion. A total of 298 cores and 355 diagnoses were obtained, with 36 (10.1%) normal prostates, 46 (13.0%) with BPH, 128 (36.1%) with PIA, 74 (20.8%) with PIN and 71 (20.0%) with PC. There is variation in the expression of uPAR in canine prostate according to the lesion, with lower expression in normal tissue and with BPH, and higher expression in tissue with PIA, PIN and PC. The high expression of uPAR in inflammatory and neoplastic microenvironment indicates increased proteolytic activity in canine prostates with PIA, PIN, and PC.(AU)
Lesões prostáticas como a neoplasia intraepitelial prostática (PIN) e a atrofia inflamatória proliferativa (PIA) são estudadas na espécie humana e canina devido ao seu potencial de malignidade. O sistema de ativador de plasminogênio (PA) tem sido sugerido como um importante mecanismo na adesão celular, angiogênese, inflamação e invasão tumoral. O receptor ativador de plasminogênio do tipo uroquinase (uPAR) é um componente do PA, expresso em células tumorais e estromais. Avaliou-se, por imunoistoquímica em lâmina de microarranjo tecidual (TMA), a expressão de uPAR no tecido prostático canino normal e com desordens proliferativas (hiperplasia prostática benigna-HPB, atrofia inflamatória proliferativa-PIA, neoplasia epitelial prostática-PIN e carcinoma-CP), com o objetivo de verificar o papel desta enzima na remodelação de matriz extracelular (ECM) e no processo de invasão tecidual. Foram obtidos 298 cores e 355 diagnósticos, sendo 36 (10,1%) próstatas normais, 46 (13,0%) com HPB, 128 (36,1%) com PIA, 74 (20,8%) com PIN e 71 (20,0%) com CP. Há variação na expressão de uPAR na próstata canina de acordo com a lesão, com menor expressão nas glândulas normais e com HPB, e maior naquelas com lesões displásicas e (PIA e PIN) e neoplásicas (CP). A superexpressão de uPAR nos microambientes inflamatório e neoplásico indica aumento da atividade proteolítica em próstatas caninas com PIA, PIN e CP.(AU)
Assuntos
Animais , Masculino , Cães , Doenças do Cão , Neoplasia Prostática Intraepitelial/enzimologia , Neoplasia Prostática Intraepitelial/veterinária , Ativador de Plasminogênio Tipo Uroquinase/análiseRESUMO
Urokinase-type plasminogen activator (uPA), a trypsin-like serine protease, has been implicated in large number of malignancies, tumor cell invasion, angiogenesis and metastasis; hence, the potent and selective inhibitors of uPA may therefore be therapeutically useful drugs for treatment of various forms of cancer. A three-dimensional quantitative structure-activity relationship (3D QSAR) study was performed on five different chemical series reported as selective uPA inhibitors employing comparative molecular field analysis (CoMFA)/comparative molecular similarity indices analysis (CoMSIA) techniques to investigate the structural requirements for substrates and derive a predictive model that may be used for the design of novel uPA inhibitors. Inclusion of ClogP did not improve the models significantly and exhibited comparable correlation coefficients with CoMFA steric and electrostatic models. 3D QSAR models were derived for 2-pyridinylguanidines (training set N = 25, test set N = 8), 4-aminoarylguanidines and 4-aminoarylbenzamidines (training set N = 29, test set N = 8), thiophene-2-carboxamindines (training set N = 64, test set N = 19), 2-naphthamidines (training set N = 32, test set N = 8), and 1-isoquinolinylguanidines (training set N = 29, test set N = 7). The CoMFA models with steric and electrostatic fields exhibited r2cv 0.452-0.722, r2ncv 0.812-0.986, r2pred 0.597-0.870, whereas CoMFA ClogP models showed r2cv 0.420-0.707, r2ncv 0.849-0.957, r2pred 0.600-0.870. The CoMSIA models displayed r2cv 0.663-0.729, r2ncv 0.909-0.998, r2pred 0.554-0.855. 3D contour maps generated from these models were analyzed individually, which provides the regions in space where interactive fields may influence the activity. Further, the predictive ability of 3D QSAR models was affirmed by predicting the activity of novel 2-naphthamidines. 3D QSAR models developed may be used in designing and predicting the uPA inhibitory activity of novel molecules.
Assuntos
Humanos , Ativador de Plasminogênio Tipo Uroquinase/análise , Ativador de Plasminogênio Tipo Uroquinase/síntese química , Ativador de Plasminogênio Tipo Uroquinase/farmacologia , Ativador de Plasminogênio Tipo Uroquinase/ultraestruturaRESUMO
Urokinase-type plasminogen activator (uPA), a trypsin-like serine protease, has been implicated in large number of malignancies, tumor cell invasion, angiogenesis and metastasis; hence, the potent and selective inhibitors of uPA may therefore be therapeutically useful drugs for treatment of various forms of cancer. A three-dimensional quantitative structure-activity relationship (3D QSAR) study was performed on five different chemical series reported as selective uPA inhibitors employing comparative molecular field analysis (CoMFA)/comparative molecular similarity indices analysis (CoMSIA) techniques to investigate the structural requirements for substrates and derive a predictive model that may be used for the design of novel uPA inhibitors. ClogP has been used as an additional descriptor in the CoMFA analysis to study the effects of lipophilic parameters on activity. Inclusion of ClogP did not improve the models significantly and exhibited comparable correlation coefficients with CoMFA steric and electrostatic models. 3D QSAR models were derived for 2-pyridinylguanidines (training set N = 25, test set N = 8), 4-aminoarylguanidines and 4-aminoarylbenzamidines (training set N = 29, test set N = 8), thiophene-2-carboxamindines (training set N = 64, test set N = 19), 2-naphthamidines (training set N = 32, test set N = 8), and 1-isoquinolinylguanidines (training set N = 29, test set N = 7). 3D contour maps generated from these models were analyzed individually, which provides the regions in space where interactive fields may influence the activity. The superimposition of contour maps on the active site of serine proteases additionally helps in understanding the structural requirements of these inhibitors. Further, the predictive ability of 3D QSAR models was affirmed by predicting the activity of novel 2-naphthamidines. 3D QSAR models developed may be used in designing and predicting the uPA inhibitory activity of novel molecules.
Assuntos
Humanos , Ativador de Plasminogênio Tipo Uroquinase/análise , Ativador de Plasminogênio Tipo Uroquinase/química , Ativador de Plasminogênio Tipo Uroquinase/farmacologia , Ativador de Plasminogênio Tipo Uroquinase/ultraestruturaRESUMO
Comparative molecular field analysis (CoMFA) and comparative molecular similarity indices analysis (CoMSIA) was performed on a series of indole/benzoimidazole-5-carboxamidines as urokinase plasminogen activator (uPA) inhibitors. The ligand molecular superimposition on template structure was performed by atom/shape-based RMS fit, multifit, and RMSD fit methods.The removal of two outliers from the initial training set of 30 molecules improved the predictivity of the models. The statistically significant model was established from 28 molecules, which were validated by evaluation of test set of nine compounds. The atom based RMS alignment yielded best predictive CoMFA model (r2cv ¬ 0:611, r2cnv ¬ 0:778, F value¬43.825, r2bs ¬ 0:842, r2pred ¬ 0:616with two components) while the CoMSIA model yielded (r2cv ¬ 0:499, r2cnv ¬ 0:976, F value¬96.36, r2bs ¬ 0:993, r2pred ¬ 0:694 with eight components). The contour maps obtained from 3D-QSAR studies were appraised for the activity trends of the molecules analyzed. The results indicate that the steric, electrostatic, and hydrogen bond donor/acceptor substituents play significant role inuPA activity and selectivity of these compounds. The data generated from the present study can be used as putative pharmacophore in the design of novel, potent, and selective urokinase plasminogen activator inhibitors as cancer therapeutics.
Assuntos
Humanos , Ativador de Plasminogênio Tipo Uroquinase/antagonistas & inibidores , Ativador de Plasminogênio Tipo Uroquinase/análise , Ativador de Plasminogênio Tipo Uroquinase , Indóis/química , Indóis/síntese química , Indóis/farmacologia , Indóis/uso terapêuticoRESUMO
We established and characterized a new mammary tumor cell line, LM2, derived from M2 mammary adenocarcinoma which spontaneously appeared in a BALB/c female mouse. The LM2 cell line has been maintained in culture for more than 40 passages and grows as poorly differentiated elongated cells. Ultrastructural and immunocytochemistry analysis revealed characteristic features of adenocarcinoma. Cytogenetic studies showed that LM2 cells are fundamentally hypotetraploid. They express metalloproteinases (MMP) and show high levels of plasminogen activator type urokinase (uPA). They were sensitive to nitric oxide (NO)-mediated cytotoxicity when NO derived from an exogenous donor. In vivo, although LM2 cells were able to grow in the lungs, they could not metastasize to the same target organ from s.c. primary tumors. The LM2 mouse mammary adenocarcinoma cell line is a suitable model to examine different aspects of tumor biology, in particular those related to the different pathways involved in the metastatic cascade and in the cytotoxicity mediated by NO.
Assuntos
Adenocarcinoma/patologia , Neoplasias Mamárias Experimentais/patologia , Células Tumorais Cultivadas , Adenocarcinoma/química , Adenocarcinoma/secundário , Aneuploidia , Animais , Feminino , Fibroblastos/patologia , Neoplasias Pulmonares/secundário , Neoplasias Mamárias Experimentais/química , Camundongos , Camundongos Endogâmicos BALB C , Microscopia Eletrônica , Invasividade Neoplásica , Metástase Neoplásica , Proteínas de Neoplasias/análise , Transplante de Neoplasias , Óxido Nítrico/farmacologia , Células Tumorais Cultivadas/química , Células Tumorais Cultivadas/efeitos dos fármacos , Células Tumorais Cultivadas/ultraestrutura , Ativador de Plasminogênio Tipo Uroquinase/análiseRESUMO
Plasma bovino tratado com sulfato de celulose produz, em placas de fibrina, múltiplas formas de ativadores de plasminogênio. A fraçäo fibrinoliticamente mais ativa foi isolada por eletroforese preparativa. Testes amidolíticos com substratos cromogênicos e testes de imunodifusäo radial com anticorpos monoclonais revelaram que o ativador de plasminogênio isolado neste trabalho possui propriedades semelhantes às da uroquinase