摘要
目的探讨脐血造血干祖细胞(hematopoietic stem-progenitor cell,HSPC)向粒-单系(colony form-ing unit-granulocyte-monocyte,CFU-GM)、红系(colony for mingunit-erythroid,CFU-E)增殖过程中HOXB6mRNA表达及全反式维甲酸(all-trans retinoic acid,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经人粒细胞-单核细胞集落刺激因子(GM-CSF)和促红细胞生成素(EPO)分别诱导后,在培养过程第3天,7天和12天的CFU-GM、CFU-E集落生成情况。②采用实时荧光定量PCR技术(fluorogenic quantitative reserve transcription polymerize chain reaction,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粒单系和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-GMHOXB6-mRNA在增殖分化的第7天表达最强烈,第12天表达明显减弱。而在CFU-E中,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-GM、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粒单系和红系祖细胞正常增殖分化过程中的调控基因之一。②ATRA能显著上调HOXB6基因的表达。
[Objective] To observe the expression level of HOXB6 gene mRNA on the differentiation and proliferation of hematopoietic stem-progenitor cell（HSPC） to colony forming unit-granulocyte-monocyte （CFU-GM） and ery hroid progenitor （CFU-E）, and whether the proliferation progress was affected by all-trans retinoic acid （ATRA）. [Methods] 10 cases of cord blood samples were offered by obstetric of the Affiliated Hospital, all the samples were collected from fetal placenta umbilical vein. There were 2 groups： ①Normal hematopoietic progenitor cell （HPC） culture was used as blank control group ②ATRA （60 nmol/L） was added into normal CFU- GM and CFU-E culture system directly as ATRA group. Observed the effect of HOXB6 gene expression after being affected by ATRA on the differentiation progress of hematopoietic stem-progenitor cell （HSPC） to CFU- GM and CFU-E was observed on the third, seventh, and twelfth day. Using fluorogenic quantitative reserve transcription polymerize chain reaction （FQ-RT-PCR） method, the expression level of HOXB6 gene during proliferation and differentiation of HSPC. The results were showed by means plus or subtracting standard deviation and increment or inhibition ratio carry through homogene tity of variance and one-way ANOVA after having statistics meaning. The means were compared between groups by ISD and SNK. [Results]①The expression of HOXB6 mRNA was detected in the two groups by real time fluoregenic quantitative reserve transcription polymerize reaction and stated by SPSS 13.0 for windows in one way ANOVA and multiple comparisons. ②During the differentiation and proliferation of hematopoietic stem-pregenitor cell to colony forming unit-granulocyte-monocyte or erythroid progenitor in vitro, the expression of HOX B6 was significantly positive. ③Compared with the expression of CFU-GM HOXB6 on day 3, the quantity of HOXB6 was obviously higher on day 7 and lower on day 12 respectively in each group. Compared with the expression of CFU-E HOXB6 on day 3, the
出处
《中国现代医学杂志》
CAS
CSCD
北大核心
2008年第11期1506-1510,共5页
China Journal of Modern Medicine
基金
四川省教育厅重点科研项目(No:2004A058)
