MLL1和ERα结合共同调控HOXA10表达的研究  

Regulation of mixed lineage leukemia histone methylases 1 collaborated with ERαon expression of HOXA10 in AML

作　　者：华川[1] 姚婕[2] HUA Chuan YAO Jie(Department of Clinical Laboratory, the 252 th Hospital of PLA , Baoding, Hebei 071000, China Department of Clinical Laboratory, Southwest Hospital, Third Military Medical University, Chongqing 400038, China)

机构地区：[1]中国人民解放军第二五二医院检验科,河北保定071000 [2]第三军医大学西南医院检验科,重庆400038

出　　处：《检验医学与临床》2017年第15期2175-2176,2179,共3页Laboratory Medicine and Clinic

基　　金：重庆市自然科学基金资助项目(CSTC2013jcyja0102)

摘　　要：目的探讨急性髓性白血病(AML)细胞中研究同源异型基因10(HOXA10)表达调控途径以及HOXA10表达作为AML防治新靶点的可能性。方法以免疫共沉淀(Co-IP)和染色质免疫共沉淀(ChIP)实验检测混合系谱白血病蛋白1(MLL1)和ERα的相互作用,以确定MLL1是通过和ERα形成蛋白质复合物结合到HOXA10的启动子上,甲基化特异PCR(MSP)检测HOXA10启动子CPG岛甲基化状态;Western-blot检测雌二醇(E2)刺激前后和MLL1表达沉默前后组蛋白H3K4甲基化状态,以明确MLL1是否通过表观遗传学途径调控HOXA10的表达;流式细胞技术检测组蛋白去甲基化酶(LSD1)处理白血病细胞前后细胞凋亡改变。结果 MLL1和ERα与HOXA10启动子上ERE区域结合在E2刺激后和MLL1表达沉默后明显增强,并且HOXA10表达明显降低,E2刺激后组蛋白H3K4甲基化状态在明显高于刺激前,且LSD1可明显诱导白血病细胞凋亡。结论MLL1通过和ERα形成蛋白复合体和HOXA10启动子序列结合并通过表观遗传学途径调控AML细胞的HOXA10基因表达,MLL1介导的HOXA10表达的改变有可能为AML的治疗提供了一个新的方向。Objective To investigate the expression regulation of homeotic genes 10（HOXA10） in pathogenesis of acute myelo- cytic leukemia（AML） cells, and try to find the regulation mechanism of HOXA10 in acute myelocytic leukemia cell lines. Methods With co-immunoprecipitation（Co-IP） and chromatin immunoprecipitation（ChIP）,the interreaction between mixed lineage leuremia histone methylases I（MLL1） and ERa was tested to confirm MLL1 bind promoter of HOXA10 through forming protein complex with ERa. MSP was used to check the methylation status of HOXA10 promoter. Western-blot was used to check the difference of H3K4 methylation status with and without estradiol（E2）,with and without MLL1 knockdown. Results MLL1 binding to EREs region of HOXA10 promoter sequence increased and HOXA10 expression was down regulated with E2 stimulation and MLL1 knockdown. LSD1 induced apoptosis of AML cells. Conclusion It has been clearly demonstrated that HOXA10 is transcriptionally reg- ulated by MLL1, which,in coordination with ERa, plays a critical role in this process with epigenetic way and suggests a potential anti-E2 treatment of AML.

关 键 词：同源异型基因10 混合系谱白血病蛋白1 ERΑ 急性髓性白血病 

分 类 号：R321.3[医药卫生—人体解剖和组织胚胎学]