全反式维甲酸对脐血红系祖细胞HOXB6基因表达的调控作用

Effect of all-trans retinic acid on the expression of HOXB6 gene in human cord blood CFU-E.

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： The objective is to observe the expression level of HOXB6 gene mRNA on the differentiation and proliferation of hematopoietic stem - progenitor cell to Erythroid progenitor （ CFU - E ）. And the proliferation progress affected by all - trans retinoic acid （ATRA）. Methods： 1. 10 cases cord blood samples were offered by obstetric of the affiliated hospital, all samples were collected from fetal placenta umbilical vein. 2. Groups ： including 2 groups. （1） Normal hematopoietic progenitor cell （HPC） culture was used as blank control group （2） ATRA （60nmol/L） was added into normal CFU - E culture system directly as ATRA group; 3. Observe the expression of HOXB6 gene on the differentiation progress of Hematopoietic Stem - Progenitor Cell （HSPC） to CFU - E affected by ATRA on the third, seventh , and twelfth day. 4. Using fluorogenic quantitative reserve transcription polymerize chain reaction （ FQ - RT - PCR ） method, and to explore the possible mechanism of ATRA up - maldevelopment to human cord blood CFU - E in genic level. 5. Statistical methods： The results were showed by means plus or subtracting standard deviation and increment or inhibition ratio Carry through homogene tity of variance ano one - way Anova after having statistics meaning. We compared means between groups by LSD and SNK. All these accomplish tics software spss 13.0. Results： 1. the mRNA expression of HOXB6 gene in the two groups was detected by real time fluorogenic quantitive reserve transcription polymerize reaction and stated by SPSS 13.0 for windows in One Way Aonva and multiple comparisons. 2. Homeobox genes （HOX） do have a regulatory function in the differentiation process of hematopoiesis. During the differentiation and proliferation of hematopoietic stem - progenitor cell to colony forming unit - granulocyte - monocyte or erythroid progenitor in vitro, the expression of HOX B6 was significant positive. 3. Compared with the expression of CFU - E HOXB6 on day 3, the quantity of HOXB6 was obviously higher on day 12 in each group. 4. The proliferation progress affected by ATRA. Compared with the expression of HOXB6 on of normal group, the expression of HOXB6 of the group ATRA were up - regulated remarkable （ P 〈 0. 05 ）. Conclusion ： Homeobox genes （HOX） do have a regulatory function in the differentiation process of hematopoiesis. Homeobox gene B6 significantly play an important role in the process which can be concerned the HOX B6 regulate the differentiation and proliferation of CFU - E. 60nmol/ml ATRA can up - regulate the expression of HOXB6.