脐血粒系祖细胞发育过程同源盒基因B7的表达及干预  

Expression and Intervention of Homeobox B7 in Development of Colony Forming Unit-Granulocyte in vitro

作　　者：张建霞[1] 刘文君[1] 

机构地区：[1]泸州医学院附属医院儿科,四川泸州646000

出　　处：《实用儿科临床杂志》2011年第15期1181-1184,共4页Journal of Applied Clinical Pediatrics

基　　金：四川省卫生厅科研课题(20060040)

摘　　要：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 observe the expression of homeobox（HOX）B7 on the differentiation and proliferation of hematopoietic stem cell（HSC） to colony forming unit-granulocyte（CFU-G） in vitro and the progress affected by all-trans-retinoic acid（ATRA）and（or）arsenic trioxide（As2O3）.And to explore the effect of ATRA and（or） As2O3 on the expression of HOXB7 gene in the development of granulocyte progenitor in genic and proteinic levels.Methods By reverse transcription polymerase chain reaction（RT-PCR）,Western blot to detect the expressions of HOXB7 mRNA and protein on the differentiation progress of HSC to CFU-G intervened by As2O3 and（or） ATRA.Results 1.Total RNA electrophoresed by agarose gel showed that the 5 S,18 S,28 S RNA strips were all explicit and intact,and had no ob-vious degradation.2.The cDNA of the HOXB7 and the ACTB gene were reservedly transcribed by RT-PCR,compared with the standard DNA Marker.They were 129 bp and 111 bp long respectively,which were in consonance with anticipation.3.HOX gene had a regulatory function in the differentiation process of hematopoiesis.The expression of HOXB7 mRNA appeared to regularly changes during the differentiation and proliferation of HSC to CFU-G in vitro in blank control group,As2O3 group,ATRA group,ATRA＋ As2O3 group,which increased slightly on the 3rd day,and were obviously higher on the 7th day,but became lower in the 12th day;compared with the expression of HOXB7 protein on the 3rd day,the quantity of HOXB7 protein was obviously higher on the 7th day and lower on the 12th day respectively in blank control group and ATRA group,there was no significant difference in HOXB7 protein at all time in As2O3 group and ATRA ＋As2O3 group.4.Compared with the expressions of HOXB7 mRNA and protein of blank control group,the expressions of HOXB7 of the ATRA group were up-regulated remarkably（P0.05）,the expression of HOXB7 of the As2O3 group were not significantly regulated（P0.05）.Conclusions HOXB7 gene may be one of a main genes in the prolif

关 键 词：粒系祖细胞 全反式维A酸 三氧化二砷 实时荧光定量聚合酶链反应 

分 类 号：R733.7[医药卫生—肿瘤]