miR-382-5p通过靶基因PTEN阻滞NB4细胞分化

miR-382-5p Blocks the Differentiation of NB4 Cells Through Targeting PTEN

该文主要研究在急性早幼粒细胞白血病NB4细胞中, miR-382-5p通过调控靶基因PTEN(phosphatase and tensin homologue)抑制了全反式维甲酸(all-trans retinoic acid, ATRA)诱导的急性早幼粒细胞分化。我们运用ATRA(1μmol/L)诱导细胞分化; Western blot检测PTEN及髓系分化标志物CD11b的蛋白质水平;实时荧光定量PCR检测miR-382-5p的表达水平;过表达PTEN的慢病毒载体分别感染NB4细胞和HL-60、THP-1细胞;脂质体转染miR-382-5p的模拟剂(mimics)和特异性抑制剂(inhibitors)NB4细胞。结果显示, PTEN促进ATRA诱导的NB4细胞分化,而在HL-60和THP-1细胞中并无明显促分化效应。NB4细胞中,脂质体转染miR-382-5p mimics在mRNA和蛋白水平均抑制了PTEN的表达,并且抑制了ATRA诱导的分化;转染miR-382-5p inhibitors则恢复了PTEN表达,同时促进了ATRA诱导的急性早幼粒细胞NB4细胞的分化。该文结果提示, miR-382-5p靶向抑制了PTEN的表达从而抑制ATRA诱导的NB4细胞分化。

In order to investigate the regulatory effects and mechanism of PTEN(phosphatase and tensin homologue) and miR-382-5p on ATRA(all-trans retinoic acid)-induced differentiation of acute promyelocytic leukemia cell line NB4.Enforced expression of PTEN in APL cells(NB4) and no-APL cells(HL-60,THP-1) was achieved through a lentivirus vector.The miR-382-5p mimic,inhibitor and negative controls were infected into NB4 cells using Lipofectamine 2000.ATRA,a typical regent was to induced granulocytic differentiation.The protein levels of CD11 b,PTEN were detected by Western blot.The mRNA expression of miRNA-382-5p and PTEN was measured by quantitative Real-time polymerase chain reaction(qRT-PCR).Data showed that overexpression of PTEN promoted ATRA-induced differentiation in APL cell line NB4 compared to no-APL line HL-60 and THP-1.Enforced expression of miR-382-5p attenuated expression of PTEN and cell differentiation marker CD11 b.Conversely,down-regulation of miR-382-5p,increased the expression of PTEN and CD11 b.It was concluded that miR-382-5p modulated ATRA-induced differentiation of APL cells by regulating its potential target PTEN.