AMANDA IKEGAMI
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Resumo IPEN-doc 23797 shRNA Knockdown of NFKB1 expression inhibits proliferation and promotes apoptosis of renal cell carcinoma2016 - IKEGAMI, A.; TEIXEIRA, L.F.; BRAGA, M.S.; SILVA, E.C. da; BELLINI, M.H.Renal cell carcinoma (RCC) represents approximately 2‐3 % of human malignancies. Despite all new therapeutic advances, almost all patients develop resistance to treatment and cure is rarely seen. The transcription factor KB (NFKB) comprises a family of transcription factors which has been associated with apoptosis resistance and progression of RCC. In this study, shRNA plasmid vector against NFKB1 gene was stably transduced into the Renca murine RCC cell line. Knockdown of NFKB1 was confirmed by quantitative real time PCR, Western blot and immunofluorescence analysis. The biological effects of decreased NFKB1 protein levels were evaluated, in vitro, by cell cycle and doubling time analysis and, in vivo, by tumor growth, cell proliferation (PCNA staining) and apoptosis (Caspase‐3 staining) and necrosis (morphometry). The results revealed that NFKB1 knockdown efficiently inhibited the growth of RencashRNA cells in culture, induced cell cycle arrest at the G2/M phase and led to a significant decrease of the doubling‐time. Moreover, NFKB1 shRNA vector suppressed tumor growth, enhanced apoptosis and necrosis compared with a wild type and mock control groups. In conclusion, our results suggest that specific silencing of NFKB is a potential therapeutic strategy for the treatment of RCC. This research was supported by FAPESP (2014/19265‐6)