siRNA干扰JARIDl1B基因表达对HL-60细胞增殖和凋亡的影响  

The effects of JARID1B siRNA on proliferation and apoptosis in HL-60 cell

作　　者：马旭东[1] 韩会丹[1] 黄轶群[1] 邹勇[1] 

机构地区：[1]福建医科大学附属漳州市医院血液科,363000

出　　处：《中华血液学杂志》2012年第5期392-396,共5页Chinese Journal of Hematology

基　　金：卫生部科学研究基金福建卫生教育联合攻关计划项目资助（wkj2008-2~55）;福建医科大学科学研究发展专项基金计划项目（FZS08018）;漳州市科学研究发展计划项目（Z07014）

摘　　要：目的研究siRNA干扰JARID1B基因表达对急性早幼粒白血病细胞系HL-60细胞增殖、凋亡的影响，并探讨其可能的机制。方法以Lipofectamine^TM2000（Lipo）为载体将JARID1B特异性siRNA转染至HL-60细胞，RT．PCR检测HL-60细胞JARIDlBmRNA的变化，Westernblot检测JAR-ID1B蛋白及组蛋白H3K4甲基化的变化；MTY法观察细胞增殖，流式细胞术检测细胞凋亡；用Westernblot分析凋亡相关蛋白Bcl-2、procaspase-9、procaspase-3、c-myc及P27的变化。结果干扰JARIDlB基因表达可上调白血病细胞组蛋白H3K4甲基化水平；siRNA干扰JARIDlB基因可抑制HL-60细胞增殖，诱导细胞凋亡；0、30、60、120nmol／LJARIDlBsiRNA处理24h后，凋亡细胞百分率分别为（11．0±3．6）％、（35．2±5．1）％、（52．7±3．8）％、（62．0±5．7）％，差异有统计学意义（F=70．27，P〈0．01）；转染后细胞Bcl-2、procaspase-9、procaspase-3、c-myc蛋白表达下降，而P27蛋白表达上升。结论JAR-ID1BsiRNA可上调组蛋白H3K4甲基化，可有效抑制肿瘤细胞增殖并诱导肿瘤细胞凋亡，其可能通过上调P27蛋白、下调Bcl-2、procaspase-9、procaspase-3、c-myc蛋白表达发挥诱导凋亡作用，有望成为白血病基因治疗的新靶点。Objective To study the effect of small interfering RNA （siRNA） targeting JARID1B gene on the proliferation and apoptosis in HL-60 cells acute promyclocytic leukemia cell line, and to explore its mechanisms. Methods The JARID1B siRNA was transfected into HL-60 cells using Lipofectamine^TM 2000 （Lipo） vector. The proliferation inhibition by siRNA targeting JARID1B was detected by MTY, cells apopto- sis by flow cytometry, the mRNA expression of JARID1B by RT-PCR, the protein expression of JARID1 B, Bcl-2, procaspase-9, procaspase-3, c-myc and P27 and histone methylated H3K4 by Western blot. Results siRNA targeting JARID1B upregulated histone methylated H3K4 level, inhibited the proliferation of HL-60 cells, and induced the cells apoptosis. After transfection of siRNA targeting JARID1B at 0, 30, 60, 120 nmol/L for 24 hours, the apoptotic rate were （11.0 ±3.6）%, （35.2 ± 5. 1）%, （52.7 ± 3.8）%, and （62.0 ± 5.7） % respectively （ F = 70.27, P 〈 0.01 ）. The protein expression of P27 was upregulated, and Bcl-2, proeaspase-9, procaspase-3, c-myc was down regulated. Conclusions JARID1B siRNA upregulates histone methylated H3K4. It reduces HL-60 cells proliferation and apoptosis by up regulating the P27 expres- sion and down regulating the Bcl-2, procaspase-9, procaspase-3, c-myc expression. It might be a new thera- peutic targeting for human leukemia.

关 键 词：基因 JARID1B RNA干扰 细胞增殖 细胞凋亡 白血病 组蛋白甲基化 

分 类 号：R363[医药卫生—病理学]