Screening and Identifying Reference Genes for Erythrocyte Production from Cord Blood CD34+Cells Exposed to Hypoxia
Screening and Identifying Reference Genes for Erythrocyte Production from Cord Blood CD34+Cells Exposed to Hypoxia
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DOI：
10.1089/dna.2023.0201
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发表时间：
2023-11-23
影响因子：
3.1
通讯作者：
Li，Cuiying
中科院分区：
文献类型：
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作者：
Xiao，Jun;Li，Zhicai;Li，Cuiying
脐带血（CB）CD34+细胞有潜力用于实现人工造血，因为它们能够在多个方向上扩增和分化。然而，这种分化的机制和分子变化仍然不清楚。CB CD34+细胞的分化通常由基因表达的细微变化驱动。检测基因表达的一个关键方法是定量实时聚合酶链反应，但结果的准确性取决于使用可靠的参考基因。本研究采用geNorm和Normalization算法对10个新候选参考基因（EIF4G2、DYNC1H1、LUC7L3、CD46、POLR1D、WSB 1、GAPVD 1、HGS、LGALS 8和RBM5）和8个传统参考基因（GAPDH、YWHAZ、ACTB、B2 MG、TBP、HMBS、PPIA和HPRT 1）在不同氧浓度下的CB CD 34+细胞转录水平进行了筛选和评价。Refresh在线工具综合分析表明，传统参考基因TBP和新型参考基因EIF4G2表达最稳定，GAPD和HMBS表达最差。这些结果可作为筛选稳定表达的参考基因的基础，以便在不同氧浓度刺激下更准确地归一化CB CD 34+细胞分化过程中。
Cord blood (CB) CD34+ cells have the potential to be used to achieve artificial hematopoiesis because of their ability to expand and differentiate in multiple directions. However, the mechanism and molecular changes underlying such differentiation are still unclear. The differentiation of CB CD34+ cells is generally driven by subtle changes in gene expression. A crucial method for examining gene expression is quantitative real-time polymerase chain reaction, but the accuracy of the results is dependent on the use of reliable reference genes. Here, the transcription levels of 10 novel candidate reference genes (EIF4G2,DYNC1H1,LUC7L3,CD46,POLR1D,WSB1,GAPVD1,HGS,LGALS8, andRBM5) and 8 traditional reference genes (GAPDH,YWHAZ,ACTB,B2MG,TBP,HMBS,PPIA,HPRT1) in CB CD34+ cells under different oxygen concentrations were screened and evaluated by using the geNorm and NormFinder algorithms. Comprehensive analysis conducted by RefFinder online tool showed thatTBP(a traditional reference gene) andEIF4G2(a novel reference gene) had the most stable expression, whereasGAPDHandHMBSwere the least suitable reference genes under these conditions. These results may serve as a basis for selecting reference genes with stable expression for more accurate normalization under different oxygen concentration stimulation during CB CD34+ cells differentiation.