Ikaros通过靶向c-KIT抑制B-ALL白血病细胞的增殖  

Ikaros Inhibits Proliferation of B-ALL Leukemia Cells by Targeting c-KIT

作　　者：宋娜[1] 贾慧婕 陈秋月 邹亚文 陈银泽 齐金博 杨俊 邱艳艳 胡庆 谷腾腾 马晓艳 王海军[2] SONG Na;IA Hui-Jie;CHEN Qiu-Yue;ZOU Ya-Wen;CHEN Yin-Ze;QI Jin-Bo;YANG Jun;QIU Yan-Yan;HU Qing;GU Teng-Teng;MA Xiao-Yan;WANG Hai-Jun(Department of Biochemistry and Molecular Biology;Department of Pathology,School of Basic Medical Sciences,Xinxiang Medical University,Xinxiang 453003,Henan,China)

机构地区：[1]新乡医学院基础医学院生物化学与分子生物学系,河南新乡453003 [2]新乡医学院基础医学院病理学系,河南新乡453003

出　　处：《中国生物化学与分子生物学报》2018年第7期778-784,共7页Chinese Journal of Biochemistry and Molecular Biology

基　　金：国家自然科学基金项目(No.81602132);国家级大学生创新创业训练计划项目(No.201610472017;201610472040);河南省科技攻关项目(No.172102310584;182102310242)资助~~

摘　　要：Ikaros是一种重要的造血细胞分化与发育调控因子,其基因结构、蛋白质活性的改变与淋巴细胞白血病的发生密切相关。致癌基因c-KIT与白血病的发生有直接联系,但Ikaros与c-KIT之间的调控关系尚未见报道。本研究报道,在人急性B淋巴细胞白血病(B-acute lymphoblastic leukemia,B-ALL)细胞中,Ikaros可靶向调控c-KIT基因的转录与蛋白质表达。通过在人B-ALL细胞系Nalm6中分别高表达和shRNA干扰Ikaros后,qRT-PCR与Western印迹结果显示,Ikaros可直接抑制c-KIT基因的表达。双荧光素酶报告实验检测Ikaros及其突变体对c-KIT基因的直接靶向作用。结果显示,野生型Ikaros可明显抑制c-KIT的表达,而突变体则不能。进一步利用染色质免疫共沉淀技术(chromatin-immunoprecipitation,Ch IP),检测Ikaros对c-KIT上游启动子序列的结合活力。结果显示,Ikaros蛋白在c-KIT的上游调控区约-500 bp处有明显的结合。Ikaros通过靶向cKIT上游启动子,抑制c-KIT表达,抑制B-ALL细胞的增殖,为临床治疗白血病提供了新思路。Ikaros is a crucial regulator of the differentiation and development of hematopoietic cells,and it is closely related to the occurrence of lymphoid leukemia. The oncogene c-KIT also has a direct relationship with leukemia,but the interaction between Ikaros and c-KIT was unknown. Here,we reported that Ikaros regulated the transcription and protein abundance of c-KIT in B-acute lymphoblastic leukemia（ B-ALL） cells. Results displayed that Ikaros could directly inhibit c-KIT expression in human B-ALL Nalm6 cells by qRT-PCR and Western blotting. Dual luciferase reporter assays were performed to determine the targeting effect of Ikaros and its mutant on the c-KIT gene. The results showed that the wild-type Ikaros could significantly inhibit the expression of c-KIT compared with the control group,but the mutant Ikaros could not. Meanwhile,Chromatin immunoprecipitation（ Ch IP） assays were performed and we found that the binding site of Ikaros on the promoter of c-KIT was at about-500 bp. We propose that Ikaros inhibits the expression of c-KIT and the proliferation of B-ALL cells by targeting the upstreampromoter of c-KIT,which provides a new venue for leukemia therapy.

关 键 词：核内转录因子Ikaros 致癌基因c-KIT 急性B淋巴细胞白血病 基因表达与调控 

分 类 号：Q819[生物学—生物工程]