转录因子Ets-1与β1,3-N-乙酰氨基葡萄糖基转移酶基因启动子结合活性分析(英文)  

Binding Analysis of Transcription Factor Ets-1 with the Upstream Sequence of β 1 ,3-N-Acetylglucosaminyltransferase Gene

作　　者：仇灏[1] 徐正荣[1] 邵雪君[2] 周迎会[1] 徐岚[1] 吴士良[1] 

机构地区：[1]苏州大学医学部生物化学与分子生物学系,苏州215123 [2]苏州大学附属儿童医院,苏州215003

出　　处：《中国生物化学与分子生物学报》2012年第1期61-65,共5页Chinese Journal of Biochemistry and Molecular Biology

基　　金：Supported by National Natural Science Foundation of China(No.30670642)~~

摘　　要：β1,3-N-乙酰氨基葡萄糖转移酶-2,-8(β3GnT-2,β3GnT-8)共同参与多聚N-乙酰氨基乳糖([Galβ1→4GlcNAcβ1→3]n)的合成,从而使得细胞表面的相应糖链结构延长进而影响细胞的恶性转化.已有研究表明,在全反式维甲酸诱导人白血病细胞株HL-60分化过程中β3GnT-2,-8的表达上调,但其分子机制不明.本文旨在探讨ATRA诱导HL-60分化过程中,转录因子Ets-1对β3GnT-2,-8表达调控的分子机制.采用10-6mol/L ATRA诱导人白血病细胞株HL-60向粒系分化,RT-PCR检测到细胞中Ets-1的表达明显增加;进一步采用染色质免疫沉淀(ChIP)结合电泳迁移率变动实验(EMSA)检测证实,有活化的Ets-1结合至β3GnT-2/-8基因调控区.以上结果表明,转录因子Ets-1对人白血病细胞株HL-60分化过程中β3GnT-2,-8基因有表达调控作用.The expression of β1,3-N-acetyl glucosamonyltransferase-2 and-8（β3GnT-2,β3GnT-8） genes,which involved in the biosynthesis of poly-N-acetyllactosamine（polyLacNAc） of cancer cells,was increased and associated with the malignant transformation.We have previously shown that the human β3GnT-2 and-8 genes were up-regulated during all-trans-retinoid acid（ATRA）-induced differentiation of human acute myeloid leukemia HL-60 cells.Here we investigated the mechanism of β3GnT-2 and-8 genes activation by the transcription factor Ets-1.RT-PCR analysis showed a significant increase of Ets-1 expression during ATRA-induced differentiation in HL-60 cells.Chromatin immuno-precipitation（ChIP） and electrophoretic mobility shift assay（EMSA） revealed Ets-1 to regulate β3GnT-2 and-8 transcription in vivo and in vitro.The results indicates that Ets-1 played an essential role in the transcriptional activation of the β3GnT-2 and β3GnT-8 genes.

关 键 词：转录因子ETS-1 β1 3-N-乙酰氨基葡萄糖转移酶 HL-60细胞系 全反式维甲酸(ATRA) 染色质免疫沉淀(ChIP) 电泳迁移率变动实验(EMSA) 

分 类 号：Q71[生物学—分子生物学]