G-CSF通过miR-146a/Smad4信号通路对HL-60白血病细胞增殖的调控作用  

Regulation of the proliferation of leukemia cells HL-60 by G-CSF via the signaling pathway of mi R-146a/Smad4

作　　者：李昕[1,2] 盛贤福 蔡佳翌[2] 徐岚[1,2] 沈莉菁[2] 钟济华[2] 陈芳源[1,2] 钟华[1,2] 

机构地区：[1]上海交通大学医学院附属仁济医院南院血液科,上海201112 [2]上海交通大学医学院附属仁济医院血液科,上海200127

出　　处：《上海交通大学学报（医学版）》2015年第12期1779-1784,共6页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家自然科学基金(81270626)~~

摘　　要：目的探讨粒细胞集落刺激因子(G-CSF)对白血病细胞株HL-60中mi R-146a/Smad4表达水平和白血病细胞增殖活性的影响。方法采用G-CSF(50 ng/m L)提前48 h处理人白血病HL-60细胞株,另设G-CSF未处理组作为对照。采用CCK8法检测细胞增殖情况,流式细胞术检测细胞周期变化,实时荧光定量PCR检测mi R-146a及Smad4 m RNA表达情况,Western blotting检测总Smad4蛋白表达。结果 G-CSF升高白血病细胞mi R-146a表达,同时降低细胞Smad4 m RNA和总Smad4蛋白的表达,提高白血病细胞的增殖活性。转染了mi R-146a过表达质粒后的白血病细胞Smad4 m RNA和总Smad4蛋白表达均降低,而用G-CSF处理转染后的细胞,其mi R-146a表达升高及Smad4表达降低得更为明显。结论 G-CSF通过调控mi R-146a/Smad4信号轴,影响Smad4表达水平,刺激处于静止状态的白血病细胞进入细胞增殖阶段。Objective To investigate the effects of granulocyte colony stimulating factor（G-CSF） on the expression level of miR-146a/Smad4 of leukemia cell line HL-60 and proliferative activity of leukemia cells.Methods Human leukemia cell line HL-60 was treated 48 h in advance and controls were untreated leukemia cells.CCK8 was applied to detect the cell proliferation and flow cytometry was used to detect the changes of cell cycle.The mRNA expressions of miR-146 a and Smad4 were detected by real-time quantitative PC R.The total protein expression of Smad4 was detected by Western blotting.Results G-CSF up-regulated the expression of leukemia cells miR-146 a,down regulated the mRNA expression and total protein expression of Smad4,and increased the proliferative activity of leukemia cells.Both mRNA expression and total protein expression of Smad4 of leukemia cells transfected by plasmids with over-expressed miR-146 a decreased.The increase of expression of miR-146 a and decrease of expression of Smad4 of transfected cells that were treated by G-CSF were more significant.Conclusion G-CSF affects the expression of Smad4 via regulating the signal pathway of miR-146a/Smad4 and stimulates leukemia cells in quiescent state entering the proliferating phase.

关 键 词：粒细胞集落刺激因子 mi R-146a/Smad4表达 细胞增殖活性 

分 类 号：R733.7[医药卫生—肿瘤]