siRNA干扰ADAR1表达对小鼠混合淋巴细胞培养的影响

Effect of siRNA-mediated silencing of ADAR1 gene on mixed mouse lymphocyte culture

目的:研究靶向ADAR1基因的siRNA在小鼠双向混合淋巴细胞培养中所扮演的角色.方法:将化学合成的靶向ADAR1基因的siRNA在Lipofectamine 2000介导下转染处于双向混合淋巴细胞培养试验中的小鼠淋巴细胞,观察转染组与未转染组细胞表型的变化;通过RT-PCR检测两组细胞中ADAR1 mRNA的变化;细胞计数和台盼蓝拒染法检测靶向ADAR1基因的siRNA对小鼠淋巴细胞增殖的抑制作用;四甲基偶氮唑蓝(MTT)法评价转染化学合成的靶向ADAR1基因的siRNA后对小鼠淋巴细胞增殖的抑制作用.结果:在经典淋巴细胞混合培养试验中,转染靶向ADAR1基因的siRNA可以明显降低小鼠淋巴细胞内ADAR1的表达,同时ADAR1-siRNA对小鼠淋巴细胞的增殖有抑制作用.结论:在经典淋巴细胞混合培养试验中,化学合成的靶向ADAR1基因的siRNA能有效抑制ADAR1基因在小鼠淋巴细胞中的表达,并对小鼠淋巴细胞增殖有抑制作用.

AIM： To investigate lymphocyte proliferation when ADAR1 is suppressed in mixed lymphocyte culture test. METHODS： Primary lymphocyte was isolated from Balb/c and C57 mice, ADAR1 siRNA was transferred into the lymphocyte cell using Lipofectamine 2000 before two-source lymphocytes were mixed and cultured. The cell morphology was observed under a light microscope, the cell proliferation was checked using cell counting, trypan blue staining and the cell viability was determined by MTT assay. The ADAR1 mRNA was detected by RT- PCR. RESULTS： When the ADAR1 expression was suppressed by the specific siRNA, the lymphocyte proliferation was inhibited in mixed lymphocyte culture test. CONCLUSION： ADAR1 has a potential role in the proliferation of mouse lymphocyte.