中文摘要：

目的构建GINS2基因siRNA真核表达质粒,并检测其对NB4细胞凋亡的影响。方法人工合成针对GINS2基因的4组siRNA干扰序列和1组无同源性的序列,测序鉴定后转染低传代人早幼粒细胞系NB4细胞,经G418筛选后,通过QRT-PCR、Western blot检测重组质粒对NB4细胞中GINS2基因mRNA转录水平和蛋白表达水平的影响;流式细胞术检测NB4细胞的凋亡情况。结果 5组重组质粒经测序证明构建正确,4组干扰质粒转染NB4细胞后,细胞中GINS2基因mRNA的转录水平和蛋白的表达水平均有所降低,其中干扰组1基因干扰率达50%,蛋白相对表达量为56%;干扰组1细胞凋亡率为32.54%,较正常对照组和NC组明显上升（P〈0.01）。结论成功构建了GINS2基因siRNA真核表达质粒,抑制GINS2基因的表达可促进NB4细胞的凋亡,为进一步研究GINS2基因在白血病中的作用奠定了基础。

英文摘要：

Objective To construct the eukaryotic expression vector for siRNA targeting GINS2 gene and investigate its effect on apoptosis of NB4 cells.Methods Four pairs of siRNA sequence targeting GINS2 gene and one pair of non-homologous sequence were synthesized,identified and transfected to human promyelocyte NB4 strain at low passage level.Positive clones were screened with G418,in which the transcription level of GINS2 mRNA and expression level of GINS2 protein were determined by QRT-PCR and Western blot respectively.The apoptosis of NB4 cells was determined by flow cytometry.Results Sequencing result proved that all the five recombinant plasmids were constructed correctly.Both the transcription level of GINS2 mRNA and expression level of GINS2 protein in NB4 cells transfected with four recombinant plasmids for siRNA targeting GINS2 gene decreased significantly.The inhibitory rates of GINS2 mRNA transcription and GINS2 protein expression in NB4 cells transfected with recombinant plasmid PGPU6 / GFP / Neo-GINS2 siRNA1 were 50% and 56% respectively.The apoptosis rate of NB4 cells transfected with recombinant plasmid PGPU6 / GFP / Neo-GINS2 siRNA1 was 32.54%,which was significantly higher than those in normal control NB4 cells and in the NB4 cells transfected with the plasmid containing non-homologous siRNA sequence（P 0.01）.Conclusion The eukaryotic expression vector for siRNA targeting GINS2 gene was constructed successfully,and the inhibition of GINS2 expression promoted the apoptosis of NB4 cells,which laid a foundation of further study on role of GINS2 gene in leukemia.