中国组织工程研究

中国组织工程研究 ›› 2011, Vol. 15 ›› Issue (32): 6041-6044.doi: 10.3969/j.issn.1673-8225.2011.32.034

• 干细胞基础实验 basic experiments of stem cells • 上一篇    下一篇

融合蛋白AML1-ETO对C/EBPα基因表达的影响

庄文越1，2，3，陈子兴1，2，赵  昀1，2，岑建农2，沈宏杰2，祁小飞2，李炳宗4   

  1. 1苏州大学医学部，江苏省苏州市  215000
    2苏州大学附属第一医院，江苏省血液学研究所，江苏省苏州市  215000
    3北华大学医学检验学院，吉林省吉林市  132013
    4 苏州大学第二附属医院血液科，江苏省苏州市215004
  • 收稿日期:2011-01-13 修回日期:2011-03-18 出版日期:2011-08-06 发布日期:2011-08-06
  • 通讯作者: 陈子兴，博士生导师，苏州大学附属第一医院，江苏省血液学研究所，江苏省苏州市215000
  • 作者简介:庄文越☆，女，1975年生，吉林省吉林市人，汉族，苏州大学医学部在读博士，主要从事血液病的表观遗传学研究。 Lzyzwy1234@yahoo.com.cn

Effects of AMLl-ETO fusion gene on the expression of C/EBPα

Zhuang Wen-yue1, 2, 3, Chen Zi-xing1, 2, Zhao Yun1, 2, Cen Jian-nong2, Shen Hong-jie2, Qi Xiao-fei2, Li Bing-zong4   

  1. 1Medical College of Soochow University, Suzhou  215000, Jiangsu Province, China
    2The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou  215000, Jiangsu Province, China
    3Medical Ecsomatics College of Beihua University, Jilin  132013, Jilin Province, China
    4Department of Hematology, the Second Affiliated Hospital of Soochow University, Suzhou  215004, Jiangsu Province, China
  • Received:2011-01-13 Revised:2011-03-18 Online:2011-08-06 Published:2011-08-06
  • Contact: Chen Zi-xing, Doctoral supervisor, Medical College of Soochow University, Suzhou 215000, Jiangsu Province, China; The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou 215000, Jiangsu Province, China
  • About author:Zhuang Wen-yue☆, Studying for doctorate, Medical College of Soochow University, Suzhou 215000, Jiangsu Province, China; The First Affiliated Hospital of Soochow University, Jiangsu Institute of Hematology, Suzhou 215000, Jiangsu Province, China; Medical Ecsomatics College of Beihua University, Jilin 132013, Jilin Province, China Lzyzwy1234@yahoo.com.cn

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摘要：

背景：已有研究显示AML1-ETO可以通过包括抑制C/EBPα的功能在内的机制引起分化的阻滞。
目的：观察AML1-ETO对髓系分化相关基因C/EBPα表达的影响，探讨其在白血病发生中的作用。
方法：应用流式细胞术检测急性单核细胞白血病细胞株U937、U937-MOCK和U937-A/E1细胞表面抗原CD11b、CD14 的表达；荧光实时定量PCR检测C/EBPα mRNA的表达；免疫印迹法检测C/EBPα蛋白的表达；染色质免疫沉淀技术研究转染细胞中AML1-ETO与C/EBPα基因启动子之间直接的相互作用情况。
结果与结论：AML1-ETO转染细胞的细胞表面分化抗原CD11b 和CD14的表达明显下降，且转染了AML1-ETO的U937细胞系中，C/EBPα的mRNA与蛋白水平均下调，转染细胞沉淀富集的DNA中含有C/EBPα基因的启动子序列。结果提示C/EBPα是AML1-ETO 的直接靶基因，AML1-ETO能下调C/EBPα的表达。

关键词: 融合蛋白, AML1-ETO, C/EBP&alpha, 基因表达, CD11b, CD14

Abstract:

BACKGROUND: Recent studies have shown that function of C/EBPα gene can be inhibited by AML1-ETO resulting in differentiation block.
OBJECTIVE: To investigate the effects of AML1-ETO fusion gene on transcription factor C/EBPα which is crucial for the differentiation of granulocytes, and to explore its role in leukemogenesis.
METHODS: The expression of cell differentiation antigen including CD11b and CD14 was detected by flow cytometry. C/EBPα gene expression was assessed by quantitative PCR, and C/EBPα protein expression was tested by Western blot. The chromatin immunoprecipitation (ChIP)-based PCR was used to investigate the direct interaction between the AML1-ETO and C/EBPα promoter in AML1-ETO positive leukemia cell line.
RESULTS AND CONCLUSION: The expression of cell differentiation antigen CD11b and CD14 significantly decreased in transfected cells. C/EBPα expression in both mRNA and protein levels decreased in U937 cells transfected with pcDNA3.1-AML1-ETO. The enriched regions in transfected cells were located within C/EBPα promoter. Results indicated that C/EBPα is the direct target gene of AML1-ETO. AML1-ETO can down-regulate the expression of C/EBPα.

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