Screening and Verification of the siRNA Targeted for the FBXL20 Gene

Xiu-hong Jia, W. Fan, Jian-chang Li
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Abstract

Objective To obtain effective siRNA fragment of HOXA10 gene and verify its function,to supply experimental evidence for tumor prevention and curation by RNAi targeting to HOXA10 gene.Methods Three pairs of small interfering RNA targeting to the different sites of HOXA10 were designed and introduced into A549.The mRNA expression of HOXA10 of A549 was detected by semi-quantitative RT-PCR,the cell proliferation was assayed by MTT,the apoptosis was measured by flow cytometry.The most effective siRNA assay was screened and was tested the relationship between it and proliferation and apoptosis.Results The mRNA of HOXA10 was inhibited by three siRNAs in A549 cells,among which siRNA1 gave the strongest inhibiting of HOXA10 by ODR was (20.190±1.698) %.The inhibitory rate of cell proliferation was (69.793±2.092) % and the apoptosis rate was (29.593±2.670) %.Conclusion siRNA1 can specifically degrade HOXA10 mRNA and inhibit the proliferation of A549 cell and promote its apoptosis. Key words: Neoplasms;  Genes, homeobox;  RNA interference;  A549 cell
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FBXL20基因靶向siRNA的筛选与验证
目的获得HOXA10基因有效siRNA片段并验证其功能,为靶向HOXA10基因的RNAi防治肿瘤提供实验依据。方法设计3对靶向HOXA10不同位点的小干扰RNA并导入A549中。半定量RT-PCR检测A549细胞HOXA10 mRNA表达,MTT检测细胞增殖,流式细胞术检测细胞凋亡。筛选最有效的siRNA检测方法,并检测其与细胞增殖和细胞凋亡的关系。结果A549细胞中HOXA10 mRNA被3种sirna抑制,其中siRNA1对HOXA10的抑制作用最强,为(20.190±1.698)%。细胞增殖抑制率为(69.793±2.092)%,细胞凋亡率为(29.593±2.670)%。结论siRNA1可特异性降解HOXA10 mRNA,抑制A549细胞增殖,促进其凋亡。关键词:肿瘤;同源框基因;RNA干扰;A549细胞
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