SILENCING OF Bc1-2 GENE BY SMALL HAIRPIN RNA INHIBITS GROWTH OF NB4 CELLS  

作　　者：何冬梅 张洹 刘革修 

机构地区：[1]Institute of Hematology, Jinan University Medical College

出　　处：《Chinese Journal of Cancer Research》2006年第4期257-260,共4页中国癌症研究（英文版）

基　　金：This work was supported by a grant from the Natural Science Program Foundation of Guangdong Province (No.04010446)

摘　　要：Objective： To investigate the effects of small hairpin RNA（shRNA） targeting Bcl-2 on the growth of NB4 cell line. Methods： Two of pairs oligonucleotides for short hairpin expression targeting the coding region of Bcl-2 mRNA were designed and chemically synthesized. Annealed oligonucleotides were inserted into Pgenesil-1 vector downstream of U6 promoter to construct RNAi plasmid. Oligonucleotide with a scrambled sequence was used as a negative control. Recombinant expression vector was identified by enzyme cutting and sequencing. Bcl-2 shRNAs were transfected into NB4 cell with Lipofectamine 2000. Western-Blot of Bcl-2 protein expression in NB4 cells was performed after transfection. The inhibition of cell growth was assessed by a MTT assay. Results： Enzyme cutting and sequencing showed that the insertion sequence was correct. Western-Blot assay showed that the expression level of Bcl-2 protein in NB4 cells decreased after Bcl-2 shRNAs treatment. There was no difference in Bcl-2 protein levels between control shRNA and untreated cells. Viable cells at 72 and 96 h after treatment with Bcl-2 shRNAs were less than that after treatment with control shRNAs and untreated NB4 cells, respectively （P〈0.05）. Control shRNA had no significant effect on the growth of the cells. Conclusion： Bcl-2 shRNA could effectively inhibit the growth of NB4 cells. Bcl-2 shRNA might be an effective anti-leukemia candidate.Objective： To investigate the effects of small hairpin RNA（shRNA） targeting Bcl-2 on the growth of NB4 cell line. Methods： Two of pairs oligonucleotides for short hairpin expression targeting the coding region of Bcl-2 mRNA were designed and chemically synthesized. Annealed oligonucleotides were inserted into Pgenesil-1 vector downstream of U6 promoter to construct RNAi plasmid. Oligonucleotide with a scrambled sequence was used as a negative control. Recombinant expression vector was identified by enzyme cutting and sequencing. Bcl-2 shRNAs were transfected into NB4 cell with Lipofectamine 2000. Western-Blot of Bcl-2 protein expression in NB4 cells was performed after transfection. The inhibition of cell growth was assessed by a MTT assay. Results： Enzyme cutting and sequencing showed that the insertion sequence was correct. Western-Blot assay showed that the expression level of Bcl-2 protein in NB4 cells decreased after Bcl-2 shRNAs treatment. There was no difference in Bcl-2 protein levels between control shRNA and untreated cells. Viable cells at 72 and 96 h after treatment with Bcl-2 shRNAs were less than that after treatment with control shRNAs and untreated NB4 cells, respectively （P〈0.05）. Control shRNA had no significant effect on the growth of the cells. Conclusion： Bcl-2 shRNA could effectively inhibit the growth of NB4 cells. Bcl-2 shRNA might be an effective anti-leukemia candidate.

关 键 词：Bcl-2 SHRNA RNAI NB4 cells 

分 类 号：R73-3[医药卫生—肿瘤]