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

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

目的研究慢病毒载体（1entivirus vector）基因转导技术对脐血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，与无基因导入的对照组细胞比较，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HNRPA0、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^＋细胞基因表达无明显影响，有望成为临床基因治疗有效且安全的工具。

Objective To investigate the influence of gene transduction mediated by lentivims 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 protein （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 23000 genes two were upregulated 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.