沉默Bmi-1表达对K562/ADM细胞体内外增殖能力的影响  

Effect of Bmi-1 Silencing on the Proliferation of K562/ADM Cells in vitro and in vivo

作　　者：刘奔 刘玉娇 刘丹丹 赵宝霞 张凯 井如男 懂思聪 孟秀香 LIU Ben;LIU Yu-Jiao;LIU Dan-Dan;ZHAO Bao-Xia;ZHANG Kai;JING Ru-Nan;DONG Si-Cong;MENG Xiu-Xiang(Pharmaceutical and Medical Technology College,Putian University,Key Laboratory of Medical Microecology(Putian University),Fujian Universities,Putian 351100,Fujian Province,China;Labaoratorial Medicine College,Dalian Medical University,Dalian 116044,Liaoning Province,China;Laboratory Center of Diagnostics,Dalian Medical University,Dalian 116044,Liaoning Province,China)

机构地区：[1]莆田学院药学与医学技术学院,医学微生态学福建省高校重点实验室,福建莆田351100 [2]大连医科大学检验医学院临床血液学教研室,辽宁大连116044 [3]大连医科大学诊断学实验中心,辽宁大连116044

出　　处：《中国实验血液学杂志》2020年第2期446-452,共7页Journal of Experimental Hematology

基　　金：辽宁省自然科学基金(2015020297);莆田学院科研项目(2019013)。

摘　　要：目的:观察沉默Bmi-1表达对慢性髓系白血病阿霉素耐药细胞株-K562/ADM细胞增殖的影响并初步探讨其分子机制。方法:将Bmi-1小干扰RNA(siRNA)序列转染到K562/ADM细胞中降低其Bmi-1的表达;采用MTT法、软琼脂集落形成实验、裸鼠成瘤实验检测沉默Bmi-1表达对K562/ADM细胞体内外增殖能力的影响;应用Western blot检测Bmi-1、PTEN、p-AKT等蛋白表达;免疫组织化学实验分析Bmi-1和Ki-67的表达情况。结果:Bmi-1基因的沉默能够抑制K562/ADM细胞的增殖活性、集落形成及裸鼠成瘤能力;Bmi-1基因沉默后,干扰组PTEN表达明显升高,而p-AKT活性明显下降;p-AKT抑制剂-LY294002处理K562/ADM细胞后,p-AKT表达降低,集落形成及裸鼠成瘤能力相比K562/ADM细胞降低;PTEN抑制剂-Bpv处理K562/ADM-S1细胞后,PTEN的表达降低,而p-AKT表达、集落形成及裸鼠成瘤能力得以重塑;Bmi-1基因沉默使得肿瘤组织中Bmi-1与Ki-67表达均下降。结论:Bmi-1基因有可能参与了对K562/ADM的体内外增殖能力的调控,PTEN/pAKT信号通路可能涉及其中。Objective:To investigate the effect of Bmi-1 gene silencing on the proliferation of K562/ADM cells in vitro and in vivo and to explore its possible molecular mechanism.Methods:The small interference RNA(siRNA)sequences of Bmi-1 were transfected into K562/ADM cells for decreasing the expression of Bmi-1.The effect of Bmi-1 silencing on the proliferation of K562/ADM cells in vitro and in vivo was detected by using MTT method,cell colony-forming test and tumoriganicity of nude mice;the expression of Bmi-1,PTEN and PAKT proteins was detected by Wertern blot.The immunohistochemistry assay was used to analyze the expression of Bmi-1 and Ki-67.Results:The Bmi-1 silencing could significantly inhibit the proliferation activity of K562/ADM cells,the cell colony-forming ability and tumorigenicity were reduced.LY294002 decreased p-AKT expression,cell colony-forming ability and tumorigenicity.Bpv reduced the PTEN expression but increased p-AKT expression and restored the colony-forming ability and tumorigenesis of K562/ADM-S1-Bpv cells.Bmi-1-siRNA dramatically suppressed the Bmi-1 and Ki-67 protein levels in xenograft tumor tissue.Conclusion:Bmi-1 can mediate the proliferation of K562/ADM cell,the PTEN/p-AKT might be involved in this pathway.

关 键 词：BMI-1 PTEN/p-AKT 增殖 白血病 

分 类 号：R733.72[医药卫生—肿瘤]