山东医药2018，Vol.58Issue(5)：1-4,4.DOI:10.3969/j.issn.1002-266X.2018.05.001
Bmi-1基因沉默对鼻咽癌细胞CNE2生物学行为的影响
Effects of silencing Bmi-1 on biological behavior of nasopharyngeal carcinoma cell line CNE2
摘要
Abstract
Objective To investigate the effects of silencing Bmi-1 gene on the biological behavior of nasopharyngeal carcinoma cell line CNE2.Methods Three primers of shRNA targeting the Bmi-1 were designed,and the recombinant plasmid PLVx-shRNA2-Bmi-1 was constructed.Virus particles were collected and the viral titer was determined after the recombinant plasmid was transfected into 293T cells.CNE2 cells were randomly divided into five groups,control group were named for untransfected lentivirus.CNE2 cells were transfected with shRNA1,2,and 3,respectively,which were named as the shRNA1 group,shRNA2 group,and shRNA3 group;cells transfected with the empty lentivirus were named as the empty vector group,and the control group was not transfected.The protein and mRNA expression levels of Bmi-1 were detected by using real-time PCR and Western blotting at 48 h after transfection and we screened out the best interference sequence with highest inhibition efficiency.Colony formation assay was used to detect ability of cell proliferation,and flow cytometry to detect the cell cycle distribution and apoptosis after silencing Bmi-1 gene.Results The sequences were inserted into the vectors which were consistent with the three designed primers,demonstrating that the recombinant lentivirus vectors containing PLVx-shRNA2-Bmi-1 were successfully constructed.The Bmi-1 shRNA 1,2,and 3 groups had lower expression of Bmi-1 mRNA and protein than that of the other two groups (both P ＜ 0.05),and the Bmi-1 shRNA 3 group had the lowest expression of Bmi-1 mRNA and protein,and had the highest inhibition efficiency (both P ＜ 0.05).Compared with the empty vector group,the proliferation increased and the apoptosis rate increased in the Bmi-1 shRNA 3 group (both P ＜ 0.05).There were higher percentage of cells in the G1 phase and lower percentage of cells in the S phase (both P ＜ 0.05).Conclusion Silencing Bmi-1 gene can inhibit the proliferation of nasopharyngeal carcinoma CNE2 cells,promote its apoptosis,and change the cell cycle.关键词
鼻咽癌/Bmi-1基因/CNE2细胞/细胞增殖/细胞凋亡/细胞周期Key words
nasopharyngeal carcinoma/Bmi-1 gene/CNE2 cell/cell proliferation/apoptosis/cell cycle分类
医药卫生引用本文复制引用
王巧,罗清,李亚军,邹彦..Bmi-1基因沉默对鼻咽癌细胞CNE2生物学行为的影响[J].山东医药,2018,58(5):1-4,4.基金项目
国家自然科学基金资助项目(81560444) （81560444）
贵州省科技厅社会发展攻关课题(黔科合SY字[2015]3046) （黔科合SY字[2015]3046）
贵州省遵义市汇川区科技局课题(20120724). （20120724）
