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目的探讨人类脐血造血干细胞向粒系祖细胞发育过程中同源盒(HOX)B7 mRNA和蛋白的表达及全反式维A酸(ATRA)和(或)三氧化二砷(As2O3)对脐血粒系祖细胞发育过程中HOXB7表达的影响。方法采用实时荧光定量PCR(RT-PCR)、Western-blot技术测定空白对照组、ATRA组、As2O3组、ATRA+As2O3组造血干细胞向粒系祖细胞增殖过程中HOXB7 mRNA及蛋白表达水平。结果 1.琼脂糖凝胶电泳显示RNA电泳图的5 S,18 S,28 S3条带型整齐,无明显拖尾及弥散,说明RNA结构完整,无明显降解。2.反转录PCR扩增各组均有目的基因HOXB7和内参照ACTB基因的cDNA产物,与DNA分子Marker相比扩增产物大小分别如下:HOXB7为129 bp,ACTB基因为111 bp,与预定理论值大小符合。3.人类造血干细胞向粒系祖细胞增殖分化过程中,空白对照组、ATRA组、As2O3组及ATRA+As2O3组的HOXB7 mRNA第3天少量表达,第7天表达较强烈,第12天表达减弱;空白对照组、ATRA组的HOXB7蛋白在增殖分化的第3天少量表达,第7天表达较高,第12天表达减弱,As2O3组及ATRA+As2O3组的HOXB7蛋白的表达在各时间点无明显差异。4.与空白对照组比较,ATRA组HOXB7 mRNA及蛋白的表达上调(P<0.05),而As2O3组HOXB7的表达无明显差异(P>0.05)。结论 HOXB7基因可能是人类造血干祖细胞向粒系祖细胞增殖分化过程的调控基因之一,HOXB7基因与粒系造血有相关性。ATRA治疗白血病的作用可能与调节HOX基因的表达有关。As2O3对HOXB7 mRNA及蛋白的表达无明显调节作用。
Objective To investigate the expression of B7 mRNA and protein of homeobox (HOX) during the development of human umbilical cord blood hematopoietic stem cells into myeloid progenitor cells and the effect of all-trans retinoic acid (ATRA) and / or As2O3 on umbilical cord blood The Influence of HOXB7 Expression on Progenitor Cell Development. Methods The expression of HOXB7 mRNA and protein during the proliferation of hematopoietic stem cells to granulocyte-derived progenitor cells in blank control group, ATRA group, As2O3 group and ATRA + As2O3 group were determined by real-time fluorescence quantitative PCR (RT-PCR) and Western- The results of agarose gel electrophoresis showed that 5S, 18S, 28S3 bands of RNA electrophoresis map were neat, no obvious tailing and dispersion, indicating that the RNA structure is intact without significant degradation. Reverse transcription PCR amplification Each group had the target gene HOXB7 and internal reference ACTB gene cDNA products, compared with the DNA molecule Marker amplification products were as follows: HOXB7 was 129 bp, ACTB gene was 111 bp, and the scheduled The theoretical value is consistent with the size. In the process of proliferation and differentiation of human hematopoietic stem cells to granulocyte progenitor cells, HOXB7 mRNA was little expressed in the blank control group, ATRA group, As2O3 group and ATRA + As2O3 group on the 3rd day, especially on the 7th day and weakened on the 12th day ; In the blank control group, the HOXB7 protein in the ATRA group was slightly expressed on the 3rd day of proliferation and differentiation, and was highly expressed on the 7th day. The expression of HOXB7 protein was decreased on the 12th day in the ATRA group and the ATRA + As2O3 group Significant differences. Compared with the blank control group, the expression of HOXB7 mRNA and protein in ATRA group was up-regulated (P <0.05), while the expression of HOXB7 in As2O3 group was not significantly different (P> 0.05). Conclusion The HOXB7 gene may be one of the regulatory genes for the proliferation and differentiation of human hematopoietic stem / progenitor cells to granulocyte-progenitor cells. HOXB7 gene is associated with granulocytic hematopoiesis. The role of ATRA in the treatment of leukemia may be related to the regulation of HOX gene expression. As2O3 has no obvious effect on the expression of HOXB7 mRNA and protein.