慢病毒载体介导的基因转导技术对人脐血CD34+细胞基因表达谱的影响

目的 研究慢病毒载体(lentivirus vector)基凶转导技术对脐血CD34+细胞基因表达的影响.方法 将携带绿色荧光蛋白(GFP)基因的第三代自身失活(self-inactivating,SIN)慢病毒载体导入CD34+细胞,提取被病毒感染细胞的总RNA,应用cDNA微阵列技术对慢病毒载体感染的脐血CD34+细胞及正常对照细胞进行差异基因表达谱分析.结果 在23000个被检测基因中,与无基因导人的对照组细胞比较,基因导入细胞中表达的上调基因共2个,分别为IGSF4和HEC基因,下调基因共6个,分别为HNRPAO、ZNF205、FLNA、CLK3、LDB1、ACTA1.进一步对筛选出的差异表达基因进行半定量RT-PCR分析证实基因表达水平变化不明显.结论 本实验中应用的慢病毒载体对脐血CD34+细胞基凶表达无明显影响,有望成为临床基因治疗有效且安全的工具.

Influence of gene transduction mediated by lentivirus vector on gene expression of human CD34+ cord blood cells

Objecfive To investigate the influence of gene transduction mediated by lentivirus vector on haman CD34+ cord blood cell(CBCs)gene expression.Methods CD34+ cells were isolated and trans-duced with the third-generation self-inactivating(SIN)lentiviral vector carrying green fluorescent protem (GFP).The total RNA from transduced cells was extracted and the differences of genotypes between the transduced and non-transduced CD34+ cells were determined with cDNA microarray analysis. Results In 23 000 genes two were upregnlated and six downregulated.These changes were not confirmed by semi-quanti-tative RT-PCR method.Conclusions Lentiviral vector used in this study do not influence significantly on the gene expression of CD34+ CBCs,and the vector system may be a useful and safe one in clinical gene therapy.