RNAi抑制Bmi-1基因表达对Jurkat细胞的影响(英文)

查看全文 作　　者：Shangen [1]Zheng;Qibin [2]Jing;Yaqiong [1]Zheng;Yinjuan [1]Ding;Qianchuan [1]Huang;Guoqiang [3]Zhao

机构地区：[1]Department of Clinical Transfusion,Wuhan General Hospital of Guangzhou Millitary Command,Wuhan 430070,China;[2]Health Care Agency of Air Force Early Institute,Wuhan 430019,China;[3]Department of Microbiology,Basic Medical Science School,Zhengzhou University,Zhengzhou 450000,China

出　　处：《The Chinese-German Journal of Clinical Oncology》索引2012年第11卷第12期,共5页

摘　　要：Objective: The aim of our study was to investigate the effects of down-regulation Bmi-1 by RNA interference (RNAi) in T Lymphocytic leukemia Jurkat cells. Methods: Two complementary oligonucleotide strands were synthesized based on the siRNA sequence targeting Bmi-1 gene. After annealing, siRNA strands were recombined into the pRNAT-U6.2 vector, and then DNA sequencing was carried out following transformation and amplification. The recombinant was transfected into Jurkat cells with liposomes. Positive colonies were obtained through G418 selection. The mRNA and protein expressions of Bmi-1 were detected by RT-PCR and Western-blot, respectively. Effects of Bmi-1 silence on cell proliferation, cell cycle and cell aging of Jurkat cells were detected by MTT assay, flow cytometry, colony formation assay and SA-β-Gal staining, respectively. Results: The siRNA recombinant targeting Bmi-1 gene was successfully constructed. All three siRNA recombinants could significantly inhibit the expression of Bmi-1. The siRNA targeting 825nt-843nt (GACCAGACCACTACT GAAT) has the strongest inhibitory effect on Bmi-1 expression, with almost complete inhibition on Bmi-1 mRNA and protein expressions. Compared with the non-transfection group and the empty vector group, growth velocity and colony formation ability were significantly decreased, while the proportion of cells in G1 phase and the percentage of senile cells were signifi-cantly increased in highly transfected group (P < 0.05). Conclusion: Down-regulation Bmi-1 by RNA interference (RNAi) could significantly inhibit the growth of Jurkat cells in vitro. 关键词：JURKAT细胞 RNA干扰 WESTERN-BLOT siRNA 脂质体转染淋巴细胞白血病 RT-PCR RNAI

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RNAi抑制Bmi-1基因表达对Jurkat细胞的影响(英文)

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