Open Access

ARTICLE

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

Amanda Ikegami^*, Luiz Felipe S. Teixeira^*, Marina S. Braga^†, Matheus Henrique Dos S. Dias^‡, Eduardo C. Lopes^‡, Maria Helena Bellini^*

* Department of Biotechnology, IPEN-CNEN/SP, São Paulo, Brazil
† Department of Pharmacy, Faculty of Pharmaceutical Sciences, University of São Paulo, São Paulo, Brazil
‡ Laboratório Especial de Toxinologia Aplicada (LETA), Center of Toxins, Immune-Response and Cell Signaling (CeTICS), Instituto Butantan, São Paulo, Brazil

Oncology Research 2018, 26(5), 743-751. https://doi.org/10.3727/096504017X15120379906339

Abstract

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-kB 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 have implicated NF-kB1 (p50) as an important molecule in tumor progression and metastasis. In the present study, a lentivirus was used to deliver shRNA targeting NF-kB1 into mouse RCC (Renca) cells. It was determined that the knockdown of the NF-kB1 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-kB1 cells have significantly diminished tumori genicity. Moreover, immunohistochemical analysis revealed an increase in necrotic areas of Renca-shRNA-NF-kB1 tumors. Thus, this study indicates that downregulation of NF-kB1 can suppress RCC tumorigenesis by inducing late apoptosis/necrosis. Therefore, NF-kB1 may be a potential therapeutic target for RCC.

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