Evaluation of angiogenic capacity of human adenocarcinoma cell line knockout for NF-ĸΒ1 protein
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2022
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ANNUAL MEETING OF THE BRAZILIAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY (SBBq), 51st; CONGRESS OF BRAZILIAN BIOPHYSICAL SOCIETY (SBBf)/LATIN AMERICAN FEDERATION OF BIOPHYSICAL SOCIETIES (Lafebs), 46th
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INTRODUCTION: Renal cell carcinoma (RCC) is the most common adult renal epithelial cancer. The most frequent
subtype of RCC is clear cell (ccRCC). Most of ccRCC patients have a mutation in the Von Hippel-Lindau (VHL) tumor
suppressor gene. The VHL gene encodes a protein, the VHL, which can up-regulate a series of intracellular proteins,
including the hypoxia inducible factor (HIF). The transcription factor NF-кB is increased in the ccRCC. OBJECTIVES:
To evaluate the impact of the NF-кB1 gene knockout on the VEGF and IL6 expression in the human RCC cells under
normoxia and hypoxia. MATERIALS AND METHODS: The CRISPR/Cas-9 technique was used to obtain 786-0 cells
knockout for the NF-кB1 protein. Western Blot assay was used to selected the clones. A hypoxia-inducing humid
chamber was used and its effectiveness was validated its effectiveness was certified by the analysis of HIF-2α
expression levels. The quantification of VEGF and IL-6 levels was measured using Real Time-PCR and MILLIPLEX
assay. DISCUSSION AND RESULTS: The VEGF gene expression in the clones was significantly lower than that
presented by the control both in normoxia (786-0-sg1 99.68±0.09%, 786-0-sg2 78.55±0.85%, 786-0-sg3
91.70±0.87%) and in hypoxia (786-0-sg1 98.30±1.49%, 786-0-sg2 75.21±4.14%, 786-0-sg3 98.44±0.18%). The
expression of IL-6 gene was also significant lower in normoxia (786-0-sg1 49.03±0.80%, 786-0-sg2 76.59±12.43%,
786-0-sg3 66.98±10.89%) and in hypoxia (786-0-sg1 95.85±0.36%, 786-0-sg2 96.45±0.49%, 786-0-sg3
91.08±1.42%). The MILLIPLEX results show that there was a significant reduction of both VEGF and IL-6 in the culture
medium of cells knocked out in normoxia and hypoxia compared to control group. CONCLUSION: Suppression of p50
expression in the clones resulted in the reduction of VEGF and IL6 in both conditions. The reduction in the IL-6 relative
expression hypoxia/normoxia demonstrates a change in cellular responsiveness to decreased levels of oxygen.
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TEIXEIRA, LUIZ F.S.; BELLINI, MARIA H. Evaluation of angiogenic capacity of human adenocarcinoma cell line knockout for NF-ĸΒ1 protein. In: ANNUAL MEETING OF THE BRAZILIAN SOCIETY FOR BIOCHEMISTRY AND MOLECULAR BIOLOGY (SBBq), 51st; CONGRESS OF BRAZILIAN BIOPHYSICAL SOCIETY (SBBf)/LATIN AMERICAN FEDERATION OF BIOPHYSICAL SOCIETIES (Lafebs), 46th, September 5-8, 2022, Águas de Lindóia, SP. Abstract... São Paulo, SP: Sociedade Brasileira de Bioquímica e Biologia Molecular - SBBq, 2022. p. 415-415. Disponível em: http://repositorio.ipen.br/handle/123456789/33907. Acesso em: 20 Apr 2024.
Esta referência é gerada automaticamente de acordo com as normas do estilo IPEN/SP (ABNT NBR 6023) e recomenda-se uma verificação final e ajustes caso necessário.