AMANDA IKEGAMI
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Artigo IPEN-doc 24344 Knockdown of NF-κB1 by shRNA inhibits the growth of renal cell carcinoma in vitro and in vivo2018 - IKEGAMI, AMANDA; TEIXEIRA, LUIZ F.S.; BRAGA, MARINA S.; DIAS, MATHEUS H. dos S.; LOPES, EDUARDO C.; BELLINI, MARIA H.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-кB 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-kB in RCC, and many 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 renal cell carcinoma (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-кB1 cells have significantly diminished tumorigenicity. Moreover, immunohistochemical analysis revealed an increase in necrotic areas of Renca-shRNANF- кB1 tumors. Thus, this study indicates that downregulation of NF-кB1 can suppress RCC tumorigenesis by inducing late apoptosis/necrosis. Therefore, NF-кB1 may be a potential therapeutic target for RCC.Artigo IPEN-doc 23209 Essential elements as biomarkers of acute kidney injury and spontaneous reversion2018 - SILVA, REGIANE M. da; KO, GUI MI; SILVA, RINALDO F.; VIEIRA, LUDMILA C.; PAULA, RAFAEL V. de; MARUMO, JULIO T.; IKEGAMI, AMANDA; BELLINI, MARIA H.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.Artigo IPEN-doc 22807 Evaluation of the applicability of Doppler velocimetry for monitoring acute kidney injury in rats2016 - SILVA, REGIANE M. da; KO, GUI M.; SANTOS, JOSE E.M.; BOIM, MIRIAN A.; IKEGAMI, AMANDA; BELLINI, MARIA H.Animal models of renal disease have been used in the study of pathogenesis and therapeutic 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.