Knockdown of NF-κB1 by shRNA Inhibits the Growth of Renal Cell Carcinoma In Vitro and In Vivo.

Renal cell carcinoma (RCC) accounts for approximately 2%-3% of human malignancies and is the most aggressive among urologic tumors. Biological heterogeneity, drug resistance, and chemotherapy side effects are the biggest obstacles to the effective treatment of RCC. The NF-κB transcription factor is one of several molecules identified to be responsible for the aggressive phenotype of this tumor. In the past decade, several studies have demonstrated the activation of NF-κB in RCC, and many have implicated NF-κB1 (p50) as an important molecule in tumor progression and metastasis. In the present study, a lentivirus was used to deliver shRNA targeting NF-κB1 into mouse RCC (Renca) cells. It was determined that the knockdown of the NF-κB1 gene led to a reduction in cell proliferation and late apoptosis/necrosis in vitro. Flow cytometry analysis demonstrated G2/M arrest in the cells. In addition, immunoblotting analysis revealed a significant increase in cyclin B1 and Bax. In vivo experiments showed that Renca-shRNA-NF-κB1 cells have significantly diminished tumorigenicity. Moreover, immunohistochemical analysis revealed an increase in necrotic areas of Renca-shRNA-NF-κB1 tumors. Thus, this study indicates that downregulation of NF-κB1 can suppress RCC tumorigenesis by inducing late apoptosis/necrosis. Therefore, NF-κB1 may be a potential therapeutic target for RCC.

Essential Elements as Biomarkers of Acute Kidney Injury and Spontaneous Reversion.

Acute kidney injury (AKI) is an important health problem and can be caused by number of factors. The use of aminoglycosides, such as gentamicin, is one of these factors. Recently, an effort has been made to find biomarkers to guide treatment protocols. Inductively coupled plasma optical emission spectroscopy (ICP-OES) was used to estimate the contents of Ca, Cu, Fe, K, Mg, Mn, Na, P, and Zn in serum and urine of the healthy, AKI, and spontaneous recovery (SR) groups of animals. The animal model of AKI and SR was validated by measuring serum and urinary urea and creatinine. The quantitative determination of the elements showed a decrease in serum levels of Ca, and Fe in the AKI group (P<0.01 vs. healthy), with a return to normal levels in the SR group, without a significant difference between the healthy and SR groups. In the urine samples, there was a decrease in P and Na levels in the AKI group (P<0.001 and P<0.01 vs. healthy), but Ca levels were increased in this group compared with the healthy and SR groups (P<0.01). These findings indicate that mineral elements might be useful as biomarkers for AKI.

Knockdown of NF-kB1 by shRNA Inhibits the Growth of Renal Cell Carcinoma In Vitro and In Vivo

Renal cell carcinoma (RCC) accounts for approximately 2%-3% of human malignancies and is the most aggressive among urologic tumors. Biological heterogeneity, drug resistance, and chemotherapy side effects are the biggest obstacles to the effective treatment of RCC. The NF-kappa B transcription factor is one of several molecules identified to be responsible for the aggressive phenotype of this tumor. In the past decade, several studies have demonstrated the activation of NF-kappa B in RCC, and many have implicated NF-kappa B1 (p50) as an important molecule in tumor progression and metastasis. In the present study, a lentivirus was used to deliver shRNA targeting NF-kappa B1 into mouse RCC (Renca) cells. It was determined that the knockdown of the NF-kappa B1 gene led to a reduction in cell proliferation and late apoptosis/necrosis in vitro. Flow cytometry analysis demonstrated G(2)/M arrest in the cells. In addition, immunoblotting analysis revealed a significant increase in cyclin B1 and Bax. In vivo experiments showed that Renca-shRNA-NF-kappa B1 cells have significantly diminished tumori genicity. Moreover, immunohistochemical analysis revealed an increase in necrotic areas of Renca-shRNA-NF-kappa B1 tumors. Thus, this study indicates that downregulation of NF-kappa B1 can suppress RCC tumorigenesis by inducing late apoptosis/necrosis. Therefore, NF-kappa B1 may be a potential therapeutic target for RCC.

Evaluation of the applicability of doppler velocimetry for monitoring acute kidney injury in rats / Avaliação da aplicabilidade da dopplervelocimetria para o monitoramento de lesão renal aguda em ratos

Animal models of renal disease have been used in the study of pathogenesis and the-rapeutic protocols. In this study, doppler ultrasound was used to evaluate dysfunction in the renal vasculature in acute kidney injury in an animal model. Eight male Wistar rats received gentamicin (80 mg/kg) for 10 days. The blood urea and creatinine levels were measured to assess renal function. All doppler ultrasound measurements were performed on both kidneys and a colour map of the renal circulation was generated. Renal function and Doppler ultrasound measurements were performed 10 days before GM treatment and on the 5th and 10th days of the assay. Gentamicin treatment led to increased serum creatinine and blood urea levels at 5 days and 10 days post initial inoculation. A significant reduction in renal artery blood flow was observed after 5 days. However, these levels remained unchanged until the 10th day, demonstrating a lack of correlation with serum creatinine and blood urea levels. Therefore, the assessment of flow blood velocity of renal arteries by doppler ultrasound is not useful for monitoring acute kidney injury in rats.(AU)

Modelos animais têm sido utilizados tanto no estudo da patogênese quanto no desenvolvimentode protocolos terapêuticos de doenças renais. No presente trabalho, aultrassonografia com doppler foi usada para avaliar disfunções na vascularização renalem um modelo animal de doença renal aguda. Oito ratos Wistar machos receberamgentamicina (80 mg/kg) durante 10 dias. Os níveis de ureia e creatinina do sangueforam medidos para avaliar a função renal. Todas as medições foram realizadasem ambos os rins, gerando um mapa da circulação renal. As medições da funçãorenal e da ultrassonografia com doppler foram feitas 10 dias antes do tratamentocom gentamicina e no 5° e 10° dia do ensaio. O tratamento levou ao aumento decreatinina sérica e ureia no sangue no 5° e 10° dia após o início da inoculação dagentamicina. Uma redução significativa no fluxo sanguíneo da artéria renal foi observadaapós 5 dias. No entanto, estes níveis permaneceram inalterados até o 10°dia, o que demonstra uma falta de correlação com os níveis de creatinina sérica eureia. Portanto, a avaliação da velocidade do fluxo sanguíneo das artérias renais, porultrassonografia com doppler não é útil para monitorar lesão renal aguda em ratos.(AU)

Evaluation of the applicability of doppler velocimetry for monitoring acute kidney injury in rats / Avaliação da aplicabilidade da dopplervelocimetria para o monitoramento de lesão renal aguda em ratos

Animal models of renal disease have been used in the study of pathogenesis and the-rapeutic protocols. In this study, doppler ultrasound was used to evaluate dysfunction in the renal vasculature in acute kidney injury in an animal model. Eight male Wistar rats received gentamicin (80 mg/kg) for 10 days. The blood urea and creatinine levels were measured to assess renal function. All doppler ultrasound measurements were performed on both kidneys and a colour map of the renal circulation was generated. Renal function and Doppler ultrasound measurements were performed 10 days before GM treatment and on the 5th and 10th days of the assay. Gentamicin treatment led to increased serum creatinine and blood urea levels at 5 days and 10 days post initial inoculation. A significant reduction in renal artery blood flow was observed after 5 days. However, these levels remained unchanged until the 10th day, demonstrating a lack of correlation with serum creatinine and blood urea levels. Therefore, the assessment of flow blood velocity of renal arteries by doppler ultrasound is not useful for monitoring acute kidney injury in rats.

Modelos animais têm sido utilizados tanto no estudo da patogênese quanto no desenvolvimentode protocolos terapêuticos de doenças renais. No presente trabalho, aultrassonografia com doppler foi usada para avaliar disfunções na vascularização renalem um modelo animal de doença renal aguda. Oito ratos Wistar machos receberamgentamicina (80 mg/kg) durante 10 dias. Os níveis de ureia e creatinina do sangueforam medidos para avaliar a função renal. Todas as medições foram realizadasem ambos os rins, gerando um mapa da circulação renal. As medições da funçãorenal e da ultrassonografia com doppler foram feitas 10 dias antes do tratamentocom gentamicina e no 5° e 10° dia do ensaio. O tratamento levou ao aumento decreatinina sérica e ureia no sangue no 5° e 10° dia após o início da inoculação dagentamicina. Uma redução significativa no fluxo sanguíneo da artéria renal foi observadaapós 5 dias. No entanto, estes níveis permaneceram inalterados até o 10°dia, o que demonstra uma falta de correlação com os níveis de creatinina sérica eureia. Portanto, a avaliação da velocidade do fluxo sanguíneo das artérias renais, porultrassonografia com doppler não é útil para monitorar lesão renal aguda em ratos.