RESUMO
Metastasis is the main cause of death from muscle-invasive urothelial carcinoma of the bladder (UCB), and the metastatic potential of tumors is often unpredictable. The role of Dachshund homolog 2 gene (DACH2) in tumorigenesis remains unexplored. We aimed to investigate whether DACH2 can be used as a biomarker to predict metastasis and prognosis of muscle-invasive UCB in a sequential training and validation fashion. For the training set (n = 40), compared with UCB patients without lymph node (LN) metastasis, both DACH2 protein and mRNA expression were greatly increased in case-matched patients with LN metastasis. For the independent validation set (n = 243), patients with primary UCB that did not express DACH2 had a longer metastasis-free survival (MFS) and overall survival (OS) than did those with tumors expressing DACH2 (5-year MFS: 88% [95% CI 80-96] versus 19% [95% CI 7-31], p < 0.001; 5-year OS: 93% [95% CI 87-99] versus 37% [95% CI 23-51], p < 0.001). Multivariable analysis of DACH2 status showed hazard ratios of 7.34 (95% CI 3.15-11.87, p < 0.001) for MFS and 3.96 (95% CI 2.04-7.16, p < 0.001) for OS which were much higher than hazard ratios associated with other independent risk factors. Collectively, DACH2 is an independent prognostic marker that can be used at initial diagnosis of UCB to identify patients who have a high potential to develop metastasis.
Assuntos
Biomarcadores Tumorais/metabolismo , Proteínas Nucleares/metabolismo , Fatores de Transcrição/metabolismo , Neoplasias da Bexiga Urinária/metabolismo , Neoplasias da Bexiga Urinária/secundário , Adulto , Idoso , Biomarcadores Tumorais/genética , Proteínas de Ligação a DNA , Feminino , Humanos , Estimativa de Kaplan-Meier , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Músculos/patologia , Invasividade Neoplásica/patologia , Proteínas Nucleares/genética , Prognóstico , Estudos Prospectivos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , RNA Neoplásico/genética , RNA Neoplásico/metabolismo , Fatores de Risco , Fatores de Transcrição/genética , Neoplasias da Bexiga Urinária/genéticaRESUMO
The aim of this study was to compare the intraoperative difference in anatomic details between loupe-assisted and microscopic varicocelectomy within the same spermatic cord. Between April 2011 and August 2011, 26 men with 33 sides containing grade 2-3 varicocele were enrolled in this study. First, one surgeon performed the open inguinal varicocelectomy under × 3.5 loupe magnification. The presumed vascular channels and lymphatics were isolated and marked without ligation. Another surgeon then microsurgically dissected and checked the same spermatic cord using an operating microscope to judge the results in terms of the ligation of the internal spermatic veins and the preservation of the arteries and lymphatics. There were significant differences in the average number of internal spermatic arteries (1.51 vs 0.97), internal spermatic veins (5.70 vs 4.39) and lymphatics (3.52 vs 1.61) between the microscope and loupe-assisted procedures (P < 0.001, P < 0.001, P < 0.001, respectively). Meanwhile, in varicocele repair with loupe magnification, an average of 1.30 ± 1.07 (43/33) internal spermatic veins per side were missed, among the overlooked veins, 1.12 ± 0.93 (37/33) were adhered to the preserved testicular artery, as well as 0.55 ± 0.79 lymphatics and 0.36 ± 0.55 arteries that were to be ligated. In conclusion, microscopic varicocelectomy could preserve more internal spermatic arteries and lymphatics and could ligate more veins than the loupe-assisted procedure. To some degree, loupe magnification is inadequate for the reliable identification and dissection of the tiny vessels of the spermatic cord, as most of the overlooked veins were adhered to the preserved testicular artery.
Assuntos
Microcirurgia/métodos , Procedimentos Cirúrgicos Urogenitais/métodos , Varicocele/cirurgia , Humanos , Ligadura , Vasos Linfáticos/cirurgia , Masculino , Cordão Espermático/irrigação sanguínea , Instrumentos Cirúrgicos , Testículo/irrigação sanguínea , Procedimentos Cirúrgicos Urogenitais/instrumentação , Procedimentos Cirúrgicos VascularesRESUMO
The mitogen-activated protein kinase (MAPK) pathway has a protective function on the management of hematologic malignancies. The aim of this study was to assess whether the induction of MAPK-mediated effects contributes to the therapeutic value of combination sorafenib and daunorubicin (DNR) treatment. Herein, we found that DNR increased phosphorylation of extracellular signal-regulated kinases (ERK1/2) in K562 cells. ERK1/2 activity was blocked by either the mitogen-induced extracellular kinase (MEK) inhibitor U0126 or a multi-kinase inhibitor sorafenib. Of note, sorafenib sensitized K562 to DNR by inhibiting proliferation and inducing apoptosis in a dose-dependent manner which was through blocking the RAF/MEK/ERK pathway. Moreover, K562 cells transfected with a constitutively active MEK2DD plasmid showed increasing IC50 values following DNR treatment compared with control cells. Combination of DNR with MEK inhibitor U0126 synergistically inhibited K562 cell growth. In conclusion, our results indicated that sorafenib sensitized K562 cells to DNR-induced cytotoxicity by downregulating p-ERK1/2 expression. DNR in combination with sorafenib may represent a new and potential therapeutic strategy in treating acute leukemia with high p-ERK1/2 levels.
Assuntos
Protocolos de Quimioterapia Combinada Antineoplásica/farmacologia , Daunorrubicina/farmacologia , MAP Quinases Reguladas por Sinal Extracelular/antagonistas & inibidores , Sistema de Sinalização das MAP Quinases/efeitos dos fármacos , Niacinamida/análogos & derivados , Compostos de Fenilureia/farmacologia , Apoptose/efeitos dos fármacos , Butadienos/farmacologia , Linhagem Celular Tumoral , Proliferação de Células , Daunorrubicina/administração & dosagem , Sinergismo Farmacológico , MAP Quinases Reguladas por Sinal Extracelular/metabolismo , Humanos , Células K562 , Proteínas Quinases Ativadas por Mitógeno/antagonistas & inibidores , Proteínas Quinases Ativadas por Mitógeno/metabolismo , Niacinamida/administração & dosagem , Niacinamida/farmacologia , Nitrilas/farmacologia , Compostos de Fenilureia/administração & dosagem , Fosforilação/efeitos dos fármacos , Transdução de Sinais/efeitos dos fármacos , Sorafenibe , Células U937 , Regulação para Cima/efeitos dos fármacosRESUMO
This study was aimed to investigate the effect of multikinase inhibitor sorafenib on the proliferation and apoptosis of U937 cells and its possible mechanism. U937 cells were treated with different concentrations of sorafenib for 48 hours. Cell viability was determined by Cell Counting Kit-8; cell apoptosis and cell ratio in cell cycle were detected by flow cytometry with Annexin V/PI staining and PI staining respectively; expressions of GSK-3ß, ß-catenin and cyclin-D1 were assayed by Western blot. The results showed that the proliferation of U937 cells was inhibited by sorafenib in a dose-dependent manner (p < 0.05). Sorafenib induced cell apoptosis and cell cycle G(1)/G(0) arrest. Compared with results of Western blot before treatment, expression of inactivated GSK-3ß, ß-catenin and Cyclin-D1 down-regulated in a dose-dependent manner after treatment with sorafenib, this same changes were observed after up-regulation of inactivated GSK-3ß by LiCl (p < 0.05). It is concluded that sorafenib inhibits the proliferation of U937 cells and induces cell apoptosis through reducing negative regulation of WNT signal pathway on inactivated GSK-3ß and down-regulating ß-catenin and cyclin-D1 level, which result in U937 cell cycle G(1)/G(0) arrest.
Assuntos
Apoptose/efeitos dos fármacos , Benzenossulfonatos/farmacologia , Piridinas/farmacologia , Via de Sinalização Wnt , Proliferação de Células/efeitos dos fármacos , Ciclina D1/metabolismo , Quinase 3 da Glicogênio Sintase/metabolismo , Glicogênio Sintase Quinase 3 beta , Humanos , Niacinamida/análogos & derivados , Compostos de Fenilureia , Sorafenibe , Células U937 , beta Catenina/metabolismoRESUMO
CpG-oligonucleotides (CpG-ODNs), mimicking bacterial DNA, have recently been shown to stimulate prostate cancer invasion in vitro via Toll-like receptor 9 (TLR9). Since cyclooxygenase 2 (COX-2), frequently overexpressed in multiple tumor types including prostate cancer, is a causal factor for tumor development, invasion and metastasis, an interesting question is raised whether TLR9 regulates COX-2 expression in prostate cancer cells. To address this question, herein we examined COX-2 expression in PC-3 cells stimulated with different doses and time courses of CpG-ODNs. The regulatory role of NF-kappaB in TLR9-mediated COX-2 expression was also investigated. CpG-ODN was found to up-regulate the expression of COX-2 in PC-3 cells in a dose- and time-dependent manner, but have little impact on COX-1 expression. Moreover, CpG-ODN also promoted nuclear translocation and activation of NF-kappaB, which appeared to be required for COX-2 induction by CpG-ODN. Overall, TLR9 up-regulates COX-2 expression in prostate cancer cells, at least partially through the activation of NF-kappaB, which may be implicated in tumor invasion and metastasis.
Assuntos
Ciclo-Oxigenase 2/genética , NF-kappa B/metabolismo , Oligodesoxirribonucleotídeos/farmacologia , Neoplasias da Próstata/enzimologia , Neoplasias da Próstata/genética , Receptor Toll-Like 9/agonistas , Regulação para Cima/efeitos dos fármacos , Linhagem Celular Tumoral , Núcleo Celular/efeitos dos fármacos , Núcleo Celular/metabolismo , Cloroquina/farmacologia , Ciclo-Oxigenase 1/genética , Ciclo-Oxigenase 1/metabolismo , Ciclo-Oxigenase 2/biossíntese , Relação Dose-Resposta a Droga , Indução Enzimática/efeitos dos fármacos , Regulação Neoplásica da Expressão Gênica/efeitos dos fármacos , Humanos , Masculino , Transporte Proteico/efeitos dos fármacos , RNA Mensageiro/genética , RNA Mensageiro/metabolismo , Fatores de TempoRESUMO
BACKGROUND AND AIMS: Recent observations suggest an implication of cyclooxygenase-2 (COX-2) in tumor lymphangiogenesis through an upregulation of vascular endothelial growth factor-C (VEGF-C) expression. However, it is unclear whether COX-2 is also associated with VEGF-C expression, tumor lymphangiogenesis and lymph node metastasis in human prostate cancer. METHODS: COX-2 and VEGF-C expression were examined in tumor tissues from 58 prostate cancer patients using immunohistochemical staining. We also analyzed the association of COX-2 and VEGF-C expression with tumor lymphangiogenesis quantified as lymphatic vessel density (LVD), lymph node metastasis, and patients' biochemical progression-free survival (b-PFS). RESULTS: High expression of either COX-2 or VEGF-C was correlated with tumor lymphangiogenesis and lymph node metastasis, as well as poor b-PFS. Moreover, a strong correlation was found between expression of COX-2 and VEGF-C (r = 0.631, p <0.001). CONCLUSIONS: COX-2 is positively associated with VEGF-C expression, tumor lymphangiogenesis and lymphatic metastasis in prostate cancer. These findings suggest that COX-2 may play a pivotal role in lymphangiogenesis and lymph node metastasis of prostate cancer via the regulation of VEGF-C expression.
Assuntos
Biomarcadores Tumorais/biossíntese , Ciclo-Oxigenase 2/biossíntese , Neoplasias da Próstata/mortalidade , Neoplasias da Próstata/patologia , Fator C de Crescimento do Endotélio Vascular/biossíntese , Idoso , Biomarcadores Tumorais/análise , Ciclo-Oxigenase 2/análise , Humanos , Linfangiogênese , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Análise de Sobrevida , Fator C de Crescimento do Endotélio Vascular/análiseRESUMO
Chronic infection and resulting inflammation promote tumor development and progression, and Toll-like receptors (TLRs) may play an important role in this process. The aim of this study was to determine whether CpG oligonucleotides (CpG-ODN), which are Toll-like receptor 9 (TLR9) agonists, can promote inflammatory cytokines release from the prostate cancer PC-3 cells through activation of nuclear factor-kappaB (NF-kappaB). Flow cytometry, semiquantitative real-time reverse transcriptase-polymerase chain reaction, enzyme-linked immunosorbent assay, and immunofluorescence analysis were used to detect the transforming growth factor-beta1 (TGF-beta1) and interleukin-8 (IL-8) release and NF-kappaB activation in PC-3 cells after CpG-ODN stimulation. CpG-ODN promoted the expression and secretion of immunosuppressive cytokines TGF-beta1 and IL-8 from PC-3 cells. In addition, after CpG-ODN stimulation, NF-kappaB nuclear translocation was also observed in PC-3 cells, contributing to CpG-induced upregulation of IL-8 and TGF-beta1. Thus, TLR9 agonists may promote IL-8 and TGF-beta1 production in human prostate cancer cells through NF-kappaB activation.