中华放射医学与防护杂志
中華放射醫學與防護雜誌
중화방사의학여방호잡지
Chinese Journal of Radiological Medicine and Protection
2012年
1期
15-19
,共5页
崔巍%尹玲玲%董凌月%李莉%刘志成%李延玲%安威%刘晓丹%杨川杰%周平坤%丁库克
崔巍%尹玲玲%董凌月%李莉%劉誌成%李延玲%安威%劉曉丹%楊川傑%週平坤%丁庫剋
최외%윤령령%동릉월%리리%류지성%리연령%안위%류효단%양천걸%주평곤%정고극
早期反应基因5%辐射敏感性%转录
早期反應基因5%輻射敏感性%轉錄
조기반응기인5%복사민감성%전록
Immediate early response gene 5%Radiation sensitivity%Transcription
目的 探讨辐射诱导早期反应基因5( Ier5)的转录调控机制.方法 运用缺失体构建、定点突变、电泳迁移率实验(EMSA)及染色质免疫共沉淀(ChIP)技术,确定宫颈癌HeLa细胞中Ier5基因启动子区域、转录因子及其结合位点.结果 发现Ier5基因的启动子区可能在缺失体Ier5-8所包含的范围(-408～ -238 bp)内.Ier5基因具有2个转录因子,分别为GCF、NFI,其中GCF的结合位点在Ier5基因启动子-388～-382 bp和-274～ -270 bp处,NFI的结合位点在Ier5基因启动子-362 ～ -357 bp处.GCF抑制Ier5基因的表达,抑制作用随γ射线吸收剂量的增加而减弱,NFI促进Ier5基因的表达.结论 Ier5基因启动子最可能的区域为-408～ -238 bp,该区域存在2个辐射敏感的转录因子GCF和NFI的结合位点.GCF对Ier5基因的顺式作用元件起负调节作用,而NFI起正调节作用.
目的 探討輻射誘導早期反應基因5( Ier5)的轉錄調控機製.方法 運用缺失體構建、定點突變、電泳遷移率實驗(EMSA)及染色質免疫共沉澱(ChIP)技術,確定宮頸癌HeLa細胞中Ier5基因啟動子區域、轉錄因子及其結閤位點.結果 髮現Ier5基因的啟動子區可能在缺失體Ier5-8所包含的範圍(-408～ -238 bp)內.Ier5基因具有2箇轉錄因子,分彆為GCF、NFI,其中GCF的結閤位點在Ier5基因啟動子-388～-382 bp和-274～ -270 bp處,NFI的結閤位點在Ier5基因啟動子-362 ～ -357 bp處.GCF抑製Ier5基因的錶達,抑製作用隨γ射線吸收劑量的增加而減弱,NFI促進Ier5基因的錶達.結論 Ier5基因啟動子最可能的區域為-408～ -238 bp,該區域存在2箇輻射敏感的轉錄因子GCF和NFI的結閤位點.GCF對Ier5基因的順式作用元件起負調節作用,而NFI起正調節作用.
목적 탐토복사유도조기반응기인5( Ier5)적전록조공궤제.방법 운용결실체구건、정점돌변、전영천이솔실험(EMSA)급염색질면역공침정(ChIP)기술,학정궁경암HeLa세포중Ier5기인계동자구역、전록인자급기결합위점.결과 발현Ier5기인적계동자구가능재결실체Ier5-8소포함적범위(-408～ -238 bp)내.Ier5기인구유2개전록인자,분별위GCF、NFI,기중GCF적결합위점재Ier5기인계동자-388～-382 bp화-274～ -270 bp처,NFI적결합위점재Ier5기인계동자-362 ～ -357 bp처.GCF억제Ier5기인적표체,억제작용수γ사선흡수제량적증가이감약,NFI촉진Ier5기인적표체.결론 Ier5기인계동자최가능적구역위-408～ -238 bp,해구역존재2개복사민감적전록인자GCF화NFI적결합위점.GCF대Ier5기인적순식작용원건기부조절작용,이NFI기정조절작용.
Objective To clarify the mechanism of immediate early response gene 5 (ler5)transcription induced by radiation. Methods Deletant construction, site-specific mutagenesis,electrophoretic mobility shift assay (EMSA) and chromatin immunoprecipitation (ChIP) were used to forecast the promoter region,binding sites and transcription factors of Ier5 gene in HeLa cells.Results The promoter region of Ier5 gene might be in the region of Ier5 -8 deletant ( -408 - -238 bp).The Ier5 gene had two transcription factors of GCF and NFI,and GCF had two binding sites located in the region of - 388 - - 382 bp and - 274 - - 270 bp of Ier5 promoter.The binding site of NFI was located in - 362 --357 bp of Ier5 promoter. GCF could inhibit the expression of Ier5 gene and this inhibition was diminished when the radiation dose increased. In contrast, NFI increased the expression of Ier5.Conclusions The most possible region of Ier5 promoter is from -408 to - 238 bp which has two binding sites for the radiosensitivity transcription factors of GCF and NFI that could negatively and positively regulate the expression of Ier5 respectively.
