论文部分内容阅读
目的探讨脐血造血干祖细胞(HSPC)向红系(CFU-E)祖细胞增殖过程中HOXB6mRNA表达及全反式维甲酸(ATRA)对HOXB6mRNA表达的影响。方法1.采用造血祖细胞体外培养技术,以ATRA持续干扰人类造血干祖细胞,观察人脐血HSPC经促红细胞生成素(EPO)诱导后,在培养过程第3天,7天和12天CFU-E集落生成情况。2.采用实时荧光定量PCR技术(FQ-RT-PCR)检测造血祖细胞增殖分化过程中HOXB6基因的表达水平。3.结果用DNA相对拷贝数和RNA表达相对量(2-△△Ct)表示HOXB6基因相对表达量。结果1.人类造血干祖细胞向红系祖细胞增殖分化过程中,各组细胞HOXB6基因均表达;2.随时间延长,CFU-E HOXB6-mRNA表达在第3天,第7天,第12天均持续表达;3.与正常对照组比较,ATRA可上调HOXB6基因的表达。结论1.在脐血CFU-E祖细胞的不同增殖阶段,HOXB6基因呈现持续稳定的表达,提示HOXB6是人类造血干祖细胞向红系祖细胞正常增殖分化过程中的调控基因之一;2.ATRA能显著上调HOXB6基因的表达。
Objective To investigate the effects of HOXB6 mRNA expression and all-trans retinoic acid (ATRA) on the expression of HOXB6 mRNA during the proliferation of progenitor cells derived from hematopoietic stem and progenitor cells (HSPC) to erythroid cells (CFU-E). Methods 1. Hematopoietic stem / progenitor cells were continuously interfered with ATRA by using hematopoietic progenitor cells culture in vitro. After induction of human umbilical cord blood HSPC by erythropoietin (EPO), the cells were cultured on the 3rd, 7th and 12th days after culture -E colony formation. Real-time fluorescent quantitative PCR (FQ-RT-PCR) was used to detect the expression of HOXB6 gene during the proliferation and differentiation of hematopoietic progenitor cells. 3. Results The relative expression of DNA and relative expression of RNA relative expression (2- △△ Ct) HOXB6 gene relative expression. HOXB6 gene was expressed in all groups of cells during the process of erythroid progenitor cell proliferation and differentiation.2.The expression of CFU-E HOXB6-mRNA on day 3, day 7, 12 Day continuous expression; 3. Compared with the normal control group, ATRA can up-regulate HOXB6 gene expression. HOXB6 gene was consistently and stably expressed at various stages of proliferation of cord blood CFU-E progenitor cells, suggesting that HOXB6 is one of the regulatory genes in the process of normal proliferation and differentiation of human hematopoietic stem / progenitor cells to erythroid progenitor cells.2. ATRA can significantly up-regulate HOXB6 gene expression.