Effect of NFBD1 gene silencing by RNA interference on radiosensitivity of nasopharyngeal carcinoma CNE-1 cells
10.3781/j.issn.1000-7431.2014.02.001
- Author:
Zhi-Hai WANG
1
Author Information
1. Department of Otorhinolaryngology
- Publication Type:Journal Article
- Keywords:
Lentivirus infection;
Nasopharyngeal neoplasms;
Radiation oncology;
RNA, small interfering
- From:
Tumor
2014;34(2):101-107
- CountryChina
- Language:Chinese
-
Abstract:
Objective: To investigate the effect of nuclear factor with BRCT domains protein 1 (NFBD1) gene silencing by short hairpin RNA (shRNA) interference on the radiosensitivity of human nasopharyngeal carcinoma CNE-1 cells. Methods: The CNE-1 cells were transfected with NFBD1-shRNA by method of lentiviral infection, the stably transfected cells were selected by puromycin, and the mRNA and protein levels of NFBD1 in CNE-1 cells were detected by real-time fluorogenic quantitative-PCR (RFQ-PCR) and Western blotting, respectively. For the examination of the effect of NFBD1 depletion on the radiosensitivity of CNE-1 cells, the flow cytometry was used to calculate the apoptotic rate and cell cycle distribution, and the survival fraction of the cells was measured via colony formation assay. Results: After transfection with NFBD1-shRNA, the mRNA and protein levels of NFBD1 in CNE-1 cells were decreased obviously. The percentage of the cells in G2/M phase and the apoptotic rate of CNE-1 cells in NFBD1-shRNA group were increased after ionizing radiation (6 MeV) as compared with those of the negative control (NC)-shRNA group (P < 0.05). The survival fraction of CNE-1 cells in NFBD1-shRNA group was significantly reduced after radiation as compared with that of the NC-shRNA group (P < 0.05). Conclusion: Lentivirus-mediated shRNA targeting NFBD1 can steadily silence the expression of NFBD1 gene and enhance the radiosensitivity of CNE-1 cells. Copyright © 2014 by TUMOR.
