转录因子Sp1抑制K562细胞红系分化  

Sp1 inhibits the erythroid differentiation of K562 cells

作　　者：马艳妮[1] 巩蓓[1] 董贺[1] 王斌[1] 阎赟梦[1] 张俊武[1] 余佳[1] 

机构地区：[1]中国医学科学院基础医学研究所医学分子生物学国家重点实验室,北京100005

出　　处：《基础医学与临床》2013年第5期557-561,共5页Basic and Clinical Medicine

基　　金：国家自然科学基金(31201103)

摘　　要：目的研究转录因子Sp1在K562细胞诱导向红系分化过程中的表达变化,并确定其对于红系分化及珠蛋白表达的影响。方法用定量PCR及Western blot的方法确定Sp1在红系分化过程中的表达情况。通过RNAi的方法抑制Sp1的表达,并通过联苯胺染色确定K562细胞中血红蛋白的表达情况,同时定量PCR的方法分析红系分化相关基因的表达,流式细胞技术检测红系分化过程中表面标志蛋白的表达情况。结果在hemin诱导的K562细胞以及促红细胞生成素EPO诱导的造血干细胞向红系分化过程中,Sp1的mRNA及蛋白水平均呈明显下降,提示其可能负调节红系分化过程。在K562细胞中抑制Sp1的表达则可明显提高K562细胞中的血红蛋白含量,促进γ-,ε-珠蛋白,CD71,CD235a基因的表达,同时CD71,CD235a阳性细胞比例明显增加。结论以上结果说明转录因子Sp1负调节红系分化过程,抑制Sp1的表达可提高K562细胞中珠蛋白的表达,并促进K562细胞向红系分化。Objective To study the expression profile of transcription factor Spl during erythroid differentiation and meanwhile to explore the function of Spl on the globin gene expression and erythroid differentaiton. Methods Q-PCR and Western blot were used to detect the expression of Spl during erythroid differentiation. Spl was knocked down through RNAi in K562 cells which were further checked by Benzidine staining. Q-PCR was per- formed to measure the hemoglobin expression. FACS was also used to identify the CD71 and CD235a positive cells. Results Both the mRNA and protein level of Spl are down-regulated during hemin-induced K562 cell and EPO-in- duced HPCs （hematopoietic progenitor cells） erythroid differentiation suggesting its negative roles in erythropoiesis. Knock down of Spl in K562 cells can improve the expression of ε-, γ-globin, CD71, CD235a and also the hemo- globin-containing cell percentage. Meanwhile, CD71 or CD235a positive cells are also increased in Spl down-regu- lating K562 cells. Conclusions Spl is a negative regulator of erythroid differentiation. Knocking down of Spl in K562 cells can improve the expression of hemoglobin meanwhile and accelerate the erythroid differentiation process.

关 键 词：SP1 K562细胞 红系分化 

分 类 号：R341[医药卫生—基础医学]