P21对HoxB4的调控机制及其影响造血干细胞增殖初步研究  

Regulatory Mechanism of P21 on HoxB4 and its Effect on Hematopoietic Stem Cell Proliferation

作　　者：李雪华 姬牧远 周艳华[3] 何志旭 舒莉萍[2,3,4] LI Xuehua;JI Muyuan;ZHOU Yanhua;HE Zhixu;SHU Liping(Department of Pediatrics,College of Clinical Medicbw,Guizhou Medical University,Guiyang 550004,Guizhou,China;Department of Immunology,School of Basic Medicine,Guizhou Medical University,Guiyang 550004,Guizhou,China;Tissue Engneering and Stern Cell Researeh Center,Guizhou Medical University,Guiyang 550004,Guizhou,China;Animal Experiment Center,Guizhou Medical University,Guiyang 550004,Guizhou,China)

机构地区：[1]贵州医科大学临床医学院儿科学教研室,贵州贵阳550004 [2]贵州医科大学基础医学院免疫学教研室,贵州贵阳550004 [3]贵州医科大学组织工程与干细胞实验中心,贵州贵阳550004 [4]贵州医科大学实验动物中心,贵州贵阳550004

出　　处：《贵州医科大学学报》2018年第6期636-640,共5页Journal of Guizhou Medical University

基　　金：国家自然科学基金项目(30960412);贵州省科技基础条件平台项目[(2009)4005];贵州省优秀科技教育人才省长专项资金(S2008-4)

摘　　要：目的:利用转基因斑马鱼模型探讨P21是否作为Hox B4的下游基因参与调控造血干细胞(HSC)的增殖发育。方法:转基因斑马鱼系Tg(z Lmo2:LDe L-Hox B4-EGFP)与Tg(z Lmo2:Cre)交配后的胚胎作为Hox B4实验组、Tg(z Lmo2:LDe L-EGFP)与Tg(z Lmo2:Cre)交配后的胚胎作为EGFP对照组、野生型斑马鱼胚胎作为空白对照组,利用P21反义mRNA探针对斑马鱼全胚胎进行原位杂交,观察P21的表达情况。结果:Hox B4实验组胚胎、EGFP实验对照组胚胎及空白对照组胚胎在时相为18 hpf、24 hpf、72 hpf时原位杂交结果未见明显区别,30 hpf及36 hpf的Hox B4实验组斑马鱼胚胎脊椎部位P21的表达发生少量下调,48 hpf的Hox B4实验组、EGFP对照组两组胚胎胸腺部位P21较空白对照组发生少量下调。结论:转基因过表达Hoxb4的斑马鱼在造血细胞增殖异常旺盛的情况下,P21表达量出现下降,提示在胚胎期造血发育过程中P21可能作为Hox B4的下游基因,并通过Hox B4负调控作用来参与调节HSC的增殖发育。Objective： To investigate the function of P21 as a downstream gene of Hox B4 to regulate the proliferation and development of hematopoietic stem cells with transgenic zebrafish models. Methods： Transgenic zebrafish Tg（ z Lmo2： LDe L-Hox B4-EGFP） and Tg（ z Lmo2： Cre） mating embryos were used as the Hox B4 experimental group,and Tg（ z Lmo2： LDe L-EGFP） and Tg（ z Lmo2： Cre） mating embryos as the EGFP control group. The wild type zebrafish embryo was used as the blank control group. P21 antisense mRNA probe was used to observe the expression of P21 in situ hybridization of zebrafish embryos. Results： There was no obvious difference in the results of in situ hybridization between the embryos of the Hox B4 experiment group and the control group in the EGFP experiment group and the blank control group in the time periods of 18 hpf,24 hpf and 72 hpf. The expression of P21 in the vertebra of zebrafish embryos in the Hox B4 experimental group of 30 hpf and 36 hpf decreased a little. In the 48 hpf Hox B4 experiment group and EGFP control group,the P21 of thymus decreased slightly compared with that in the blank control group. Conclusions： In the case of abnormal proliferation of hematopoietic cells,the expression of P21 in zebrafish with overexpressed Hox B4 decreases. It is suggested that P21 may serve as the downstream gene of Hox B4 during the development of hematopoiesis and the negative regulation of Hox B4 to regulate the proliferation and development of hematopoietic stem cells（ HSC）.

关 键 词：造血干细胞 斑马鱼 转基因 细胞增殖 P21 HOXB4 

分 类 号：R321-33[医药卫生—人体解剖和组织胚胎学]