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机构地区:[1]蚌埠医学院临床检验诊断学实验中心,安徽蚌埠233030 [2]蚌埠医学院免疫学教研室,安徽蚌埠233030
出 处:《蚌埠医学院学报》2010年第8期757-761,共5页Journal of Bengbu Medical College
基 金:安徽省教育厅自然科学研究重点资助项目安徽省教育厅自然科学(KJ2007A097)
摘 要:目的:观察4-1BBL基因小分子干扰RNA(siRNA)对HL-60细胞生长及诱导其凋亡作用的影响。方法:化学合成法合成靶向人4-1BBL基因siRNA,脂质体转染法转染HL-60B细胞;半定量PCR和流式细胞术检测转染前后HL-60B细胞4-1BBL的表达水平,MTT法检测HL-60B细胞增殖,流式细胞术检测细胞凋亡。结果:靶向4-1BBL基因siRNA转染HL-60B细胞后,4-1BBLmRNA和蛋白的表达明显降低;细胞增殖被抑制,其抑制作用在转染后48h最明显,增生抑制率达到(22.1±2.3)%;细胞凋亡率增加,由(9.8±2.5)%上升到(32.5±5.1)%。结论:化学合成的siRNA在体外能成功抑制靶基因4-1BBL的表达和HL-60B细胞的增殖,并有助于细胞趋向凋亡。Objective:To observe the influence of 4-1BBL small interfering RNA(siRNA) on the growth and apoptosis of HL-60B cells.Methods:The siRNA for 4-1BBL gene was designed and synthesized in vitro,and then transfected into HL-60B cells by Lipofectamine;the expression of 4-1BBL mRNA and protein was detected by RT-PCR and flow cytometry,respectively;the proliferative rate of HL-60B cells was determined by methyl thiazoy tetralium(MTT),and the cell apoptosis was analyzed by flow cytometry.Results:After the siRNA for 4-1BBL gene was transfected into HL-60B cells,the expression of 4-1BBL gene and protein decreased,the growth of cells was repressed and the proliferation rate of HL-60B cells decreased.The inhibition rate was(22.1±2.3) % 48 h after transfection;the apoptosis rate increased from(9.8±2.5) % to(32.5±5.1) %.Conclusions:Chemically synthetic siRNA can decrease 4-1BBL gene expression,inhibit cellular proliferation and promote apoptosis in vitro.
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