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Renal cell carcinoma (RCC) is a malignant tumor originating from renal tubular epithelial cells with poor prognosis and high metastatic rate. Tripartite motif-containing 24 (Trim24) is a member of the tripartite motif (Trim) family and also a valuable oncogene, but its role in RCC remains unclear. We constructed the overexpression and knockdown of Trim24 cell lines to investigate its roles in RCC progression. CCK8, wound healing, and transwell assay were performed to determine the proliferation, migration, and invasion of RCC cell lines, respectively. Moreover, the expression of Trim24 and its clinicopathological significance were evaluated in a human RCC tissue microarray. From our results, Trim24 promoted the proliferation, migration, and invasion of RCC cells in vitro. Importantly, overexpression of Trim24 led to a significant increase in the expression levels of MMP-2, MMP-9, fibronectin, snail, vimentin, N-cadherin, and ß-catenin, inducing the EMT process in turn, while the expression of these proteins was significantly downregulated when Trim24 was knocked down in ACHN cells. In addition, Trim24 was significantly upregulated in RCC, and its high expression was negatively associated with the tumor size. Trim24 might operate as an oncogene in RCC progression by inducing the EMT process, suggesting that Trim24 was a potential target for human RCC.
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OBJECTIVES: We investigated the relationship between the expression of tumor necrosis factor-inducible gene 6 (TSG-6) with inflammation and integrity of the bladder epithelium in the bladder tissues of patients with bladder pain syndrome/interstitial cystitis (BPS/IC) and the mechanism of action using a rat model of BPS/IC. MATERIALS AND METHODS: Expression of TSG-6 and uroplakin III was determined by immuno- histochemistry of bladder biopsy samples from control human subjects and patients with verified BPS/IC. Our rat model of BPS/IC was employed to measure the perfusion of bladders with hyaluronidase, and assessment of the effect of TSG-6 administration on disease progression. Treatment effects were assessed by measurement of metabolic characteristics, RT-PCR of TGR-6 and interleukin-6, bladder histomorphology, and immunohistochemistry of TGR-6 and uroplakin III. RESULTS: The bladders of patients with BPS/IC had lower expression of uroplakin III and higher expression of TSG-6 than controls. Rats treated with hyaluronidase for 1 week developed the typical signs and symptoms of BPS/IC, and rats treated with hyaluronidase for 4 weeks had more serious disease. Administration of TSG-6 reversed the effects of hyaluronidase and protected against disease progression. CONCLUSION: Our results indicate that TSG-6 plays an important role in maintaining the integrity of the bladder epithelial barrier.
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OBJECTIVE: To investigate the expressions of interleukin-17 (IL-17) and interleukin-8 (IL-8) in benign prostatic hyperplasia (BPH) and BPH complicated with histological inflammation and their significance. METHODS: According to the results of HE staining, we divided 60 cases of BPH treated by transurethral resection of the prostate (TURP) into a BPH group (n = 23) and a BPH with inflammation group (n = 37). We analyzed the clinical data of the patients and determined the mRNA and protein expressions of IL-17 and IL-8 by immunohistochemistry, real-time fluorescence quantitative PCR, and Western blot, respectively. RESULTS: Compared with the BPH patients complicated with inflammation, the BPH group showed significantly lower International Prostate Symptom Score (IPSS) (29.1 ± 6.2 vs 21.6 ± 3.7), quality of life score (QoL) (5.4 ± 1.3 vs 4.4 ± 1.6), postvoid residual urine volume (RUV) (ï¼»198.6 ± 15.5ï¼½ vs ï¼»98.2 ± 19.3ï¼½ ml), prostate volume (ï¼»69.2 ± 24.1ï¼½ vs ï¼»49.8 ± 16.5ï¼½ ml), PSA level (ï¼»7.4 ± 1.9ï¼½ vs ï¼»2.8 ± 0.8ï¼½ µg/L) and serum c-reactive protein content (CRP) (ï¼»5.1±2.0ï¼½ vs ï¼»1.5±0.6ï¼½ mg/L), but a higher maximum urine flow rate (Qmax) (ï¼»4.7 ± 2.1ï¼½ vs ï¼»8.2 ± 1.8ï¼½ ml/s) (all P<0.05). The former group had a significantly higher incidence rate of urinary retention than the latter (32.4% ï¼»12/37ï¼½ vs 8.69% ï¼»2/23ï¼½), mRNA expressions of IL-17 (0.303 ± 0.076 vs 0.042 ± 0.019) and IL-8 (0.536 ± 0.059 vs 0.108 ± 0.025), and protein expressions of IL-17 (0.88 ± 0.10 vs 0.34 ± 0.05) and IL-8 (1.07 ± 0.08 vs 0.43 ± 0.04) (all P<0.05). CONCLUSIONS: The expressions of IL-17 and IL-8 are upregulated in the prostatic tissue of the BPH patients with inflammation, which may play a significant role in the development and progression of BPH.
Assuntos
Interleucina-17/metabolismo , Interleucina-8/metabolismo , Hiperplasia Prostática/metabolismo , Progressão da Doença , Humanos , Inflamação/metabolismo , Masculino , Tamanho do Órgão , Próstata/patologia , Hiperplasia Prostática/complicações , Qualidade de Vida , RNA Mensageiro/metabolismo , Ressecção Transuretral da Próstata , Resultado do Tratamento , Retenção Urinária/diagnóstico , Retenção Urinária/etiologiaRESUMO
OBJECTIVE: To investigate the changes of the expressions of the nerve growth factor (NGF) and its mRNA in the prostate tissues of spontaneously hypertensive rats (SHR) of different ages and their significance. METHODS: SHRs and normotensive Wistar Kyoto (WKY) rats were killed at 1 month (young), 6 months (adult) and 12 months (aging), respectively, 5 in each group. Their prostate indexes were calculated, and the expressions of NGF and its mRNA in the ventral prostate tissue were detected by immunohistochemistry and RT-PCR. RESULTS: The prostate indexes of the SHR and WKY groups were 1.16 +/- 0.06 and 1.03 +/- 0.09 at 1 month, 1.12 +/- 0.14 and 0.93 +/- 0.07 at 6 months, and 1.11 +/- 0.05 and 0.96 +/- 0.09 at 12 months, significantly higher in the former group than in the latter either at 6 or at 12 months (P < 0.05), but with no obvious difference at 1 month (P > 0.05). The expressions of NGF and its mRNA in the ventral prostate tissue were detected in all groups, and elevated gradually with the increase of age (P < 0.05). Among those of the same age, the expression levels were markedly higher in the SHR than in the WKY group (P < 0.05). CONCLUSION: In SHRs with benign prostate hyperplasia (BPH), the enhanced excitation of the sympathetic nervous system may be a common mechanism underlying BPH and hypertension, and NGF plays an important role in it.
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Fatores de Crescimento Neural/metabolismo , Próstata/metabolismo , Envelhecimento , Animais , Masculino , Próstata/patologia , Hiperplasia Prostática/metabolismo , Ratos , Ratos Endogâmicos SHR , Ratos Endogâmicos WKYRESUMO
OBJECTIVE: To investigate the expression of COX-2 and VEGF in clear cell renal cell carcinoma (CCRCC) and their correlation with tumor angiogenesis. METHODS: Envision immunohistochemistry was used to determine the expression of COX-2 and VEGF, and microvessel density (MVD) was marked by CD34 in 80 CCRCC tissues and 20 normal kidney tissues. The relationship between the above mentioned markers were analyzed. RESULTS: Expressions of COX-2 and VEGF were noted in both CCRCC and normal kidney tissues. The positive rates of COX-2 and VEGF were significantly higher in CCRCC than in normal kidney (P < 0.05); The expression of COX-2 was correlated with TNM stage (P < 0.05), histological grade (P < 0.05) and lymph node metastasis (P < 0.05) in CCRCC, but not with age (P = 0.663) and diameter of tumor (P = 0.528). Both COX-2 expression (r = 0.851, P < 0.01) and VEGF expression (r = 0.736, P < 0.01) were significantly associated with MVD in CCRCC. There was a positive correlation between expression of cox-2 and that of VEGF in CCRCC. CONCLUSION: COX-2 expression is correlated with tumor angiogenesis in CCRCC. It is likely that VEGF is one of the most important mediators in the COX-2 angiogenic pathway.
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Carcinoma de Células Renais , Ciclo-Oxigenase 2/metabolismo , Neoplasias Renais , Microvasos/patologia , Fator A de Crescimento do Endotélio Vascular/metabolismo , Adulto , Idoso , Idoso de 80 Anos ou mais , Carcinoma de Células Renais/metabolismo , Carcinoma de Células Renais/patologia , Feminino , Humanos , Neoplasias Renais/metabolismo , Neoplasias Renais/patologia , Metástase Linfática , Masculino , Pessoa de Meia-Idade , Estadiamento de Neoplasias , Neovascularização Patológica/metabolismo , Neovascularização Patológica/patologiaRESUMO
OBJECTIVE: To explore the feasibility and safety of gene transfer into porcine myocardium via the pericardial cavity by a homemade easy device. METHODS: Replication-deficient recombinant adenoviral vector carrying LacZ report gene (Ad-LacZ) was constructed by the calcium phosphate precipitation method. Twelve healthy Chinese mini-swine were randomly divided into experimental group (n = 6) and control group (n = 6). Acute myocardial infarction (AMI) model was established by balloon occlusion of the distal part of D1 branch of left anterior descending (LAD) artery, at the same time the intrapericardial cavity injections were performed through the small incision of the abdominal wall below the xyphoid appendix using a homemade device. Then gene transfer was performed using a central venous catheter. The pericardium was pretreated with injection of a mixture of collagenase (1,200 U) and hyaluronidase (3,000 U) in both groups. Then 2.0 x 10(9) plaque formation unit (PFU) Ad-LacZ was injected into the pericardial cavity in experimental group, while 1 mL of normal saline was injected in the control group. The beta-galactosidase activity detection and X-gal staining of the ischemic myocardium were performed on the 3rd, 7th, and 28th day after injection. RESULTS: The LAD artery was occluded completely and infarction and ischemia were detected by histological assessment In experimental group, the X-gal staining positive cells and beta-galactosidase activity quantification were detectable on the 3rd day after injection, increased markedly on the 7th day, and then declined on the 28th day. The transfer efficiencies indicated by the positive myocardial cells were 16.7%, 45.6% , 22.8% on the 3rd, 7th, 28th day, respectively. In control group, no positive cells and beta-galactosidase activity were observed. CONCLUSION: Adenovirus can be transferred into ischemic myocardium and express target gene in the AMI model for four weeks with the homemade easy device via pericardial cavity pretreated by collagenase and hyaluronidase.
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Técnicas de Transferência de Genes , Coração , Óperon Lac , Pericárdio/fisiologia , Punções/métodos , Adenoviridae/genética , Animais , Animais Geneticamente Modificados , Genes Reporter , Vetores Genéticos , Miocárdio/enzimologia , Suínos , Porco Miniatura , beta-Galactosidase/genéticaRESUMO
OBJECTIVE: In diabetes, intracellular accumulation of sorbitol resulting from the high extracellular levels of glucose leads to depletion of intracellular compounds including taurine. This is associated with the development of late diabetic complications such as cardiomyopathy. The development of myocyte hypertrophy has been largely attributed to angiotensin II, whose growth properties are antagonized by taurine. However, the interaction between taurine, angiotensin II type2 receptor (AT2) and cardiomyopathy related to angiotensin II is still unknown. This study investigates the roles of taurine and AT2 in rats with streptozotocin (STZ)-induced diabetic cardiomyopathy. METHODS: Of 60 female 4-week-old Wistar rats, 8 were treated with common diet and the other 52 with high sugar/fat diet (during the whole experiment) to induce insulin resistance. At the 4th week, of the 52 rats, 7 treated with sodium citrate buffer (pH = 4.5) were grouped into control group1 (con1) and the other 45 were treated by intraperitoneal injection (I.P) with STZ to develop type 2 diabetes. At the 28th week, the maximal velocity decrease of pressure per second in left ventricle within the period of isovolumic relaxation (-dp/dt(max)) was detected by a cannula through right carotid artery. After the cannula operation, of the 45 rats, all the living 24 with -dp/dt(max)< or = 5250 mmHg/s, who had developed diabetic cardiomyopathy, were grouped as follows: 7 treated with double distilled H2O (I.P) were grouped into control group2 (con2). 8 treated with AT2 agonist (CGP42112A) (I.P) were grouped into experimental group1 (exp1). Another 9 treated with taurine (I.P) were grouped into experimental group2 (exp2). All injections lasted 4 weeks (Q.D) and the heart weight (HW) was recorded. To examine cardiomyocyte apoptosis index (CAI), mRNA and protein of AT2 and Bcl-2 in cardiomyocytes, methods of terminal-deoxynucltidyl transferase mediated nick end labeling (TUNEL), reversal transcription polymerase chain reaction (RT-PCR) and immunoblot (Western Blot) were used, respectively. RESULTS: Values of -dp/dt(max) in exp1, exp2 or con2 were much less than those in con1, respectively (p < 0.01). CAI (= stained cell number/total cell number x 100%) and AT2 values both in mRNA and protein levels in con1 were less than those in the other three groups, respectively (p < 0.01). The three parameters above were more in exp1 but less in exp2 than those in con2, respectively (p < 0.01). The three parameters and HW in exp1 were much higher than those in exp2, respectively (p < 0.01). Changes of Bcl-2 were opposed to those of AT2. CONCLUSIONS: A high expression of AT2 may accelerate the apoptosis of cardiomyocytes in diabetic rats and play a role in precipitating diabetic cardiomyopathy; taurine may protect diabetic rats from developing cardiomyopathy also by downregulating AT2 receptors.
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Cardiomiopatias/prevenção & controle , Complicações do Diabetes/prevenção & controle , Diabetes Mellitus Tipo 2/tratamento farmacológico , Regulação para Baixo/efeitos dos fármacos , Receptor Tipo 2 de Angiotensina/biossíntese , Receptor Tipo 2 de Angiotensina/efeitos dos fármacos , Taurina/uso terapêutico , Animais , Apoptose/fisiologia , Biomarcadores/sangue , Glicemia/metabolismo , Western Blotting , Cardiomiopatias/complicações , Cardiomiopatias/patologia , Artéria Carótida Interna/metabolismo , Complicações do Diabetes/complicações , Complicações do Diabetes/patologia , Diabetes Mellitus Experimental/complicações , Diabetes Mellitus Experimental/tratamento farmacológico , Diabetes Mellitus Experimental/patologia , Diabetes Mellitus Tipo 2/complicações , Diabetes Mellitus Tipo 2/patologia , Modelos Animais de Doenças , Feminino , Imuno-Histoquímica , Insulina/sangue , Modelos Cardiovasculares , Miócitos Cardíacos/metabolismo , Tamanho do Órgão , Proteínas Proto-Oncogênicas c-bcl-2/biossíntese , RNA Mensageiro/metabolismo , Ratos , Ratos Wistar , Reação em Cadeia da Polimerase Via Transcriptase Reversa , Volume Sistólico/fisiologiaRESUMO
To study the angiogenic potency of hypoxia-prestimulated bone marrow stromal cells (BMSCs) when transplanted into acute myocardial infarction models of rats. BMSCs were cultured under hypoxia condition for 24 h. Their expression of VEGF was investigated. The rat acute myocardial infarction models were made by coronary artery ligation and divided into 3 groups at random. In normoxia group, twice-passaged BMSCs were labeled with Bromodeoxyuridine (BrdU) and then implanted into the infarction regions and ischemic border of the recipients in 4 weeks. The rats in hypoxia group were implanted with hypoxia-prestimulated BMSCs. In control group, the model rats received only DMEM medium injection. Six-weeks after AMI, the infarction regions were examined to identify the angiogenesis and the expression of the VEGF. Our results showed that viable cells labeled with BrdU could be identified in the host hearts. The infarction regions in normoxia and hypoxia groups had a greater capillary density and increased VEGF expression than the regions in control group. The capillary density and VEGF expression in hypoxia group were higher than in normoxia group. It is concluded that the enhanced expression of VEGF in BMSCs could be induced by ex vivo hypoxia stimulation. BMSCs implantation promoted the angiogenesis in myocardial infarction tissue via supplying exogenic VEGF. Angiogenic potency of bone marrow stromal cells was improved by ex vivo hypoxia prestimulation though the enhanced VEGF expression.