Role and molecular mechanism of STAT1 signaling pathway in the differentiation of leukemia cells into granulocytes induced by all-trans retinoic acid
SUN Linlin
GUO Qiang
ZHU Xiaoxiao
ZHANG Zhen
ZHAO Lin
WEI Ran
SHI Yin
YIN Xunqiang
ZHANG Yunhong
JIANG Guosheng
LI Xia
Abstract：Objective To detect the role and regulatory mechanism of signal transducer and activator of transcription 1 (STAT1) in promoting the differentiation of human leukemia cells into granulocytes induced by all-trans retinoic acid (ATRA),and to provide a novel target for clinical treatment.Methods Human acute myeloid leukemia cells HL-60 were randomly divided into the control group and observation group.In the observation group,HL-60 cells were induced to differentiate into granulocytics with ATRA (2 μmol/L),while cells in the control group were given the same concentration of DMSO.Morphologic changes of cells were observed by Wright-Giemsa staining.Cell surface marker CD11b was detected by flow cytometry.The mRNA expression levels of STAT1 and CCAAT/enhancer binding proteinsε (C/EBPε) were detected by q-PCR,the protein and phosphorylation levels of STAT1 (p-STAT1) were detected by Western blotting,and the transcriptional level of miR-155-5p was detected by q-PCR.Pearson correlation analysis was used to detect the correlation of STAT1 with C/EBPε and miR-155-5p.Results Compared with the control group,the HL-60 cells in the observation group became smaller and irregular,cytoplasm increased,nuclear concentrated,nucleolus became smaller or disappeared,and the number of leaf nuclei increased significantly.The rate of CD1 1b positive HL-60.cells in the observation group at 96 h after ATRA induction was 62.97％ ± 1.90％,which was higher than that in the control group (1.87％ ±0.08％) (P ＜ 0.05).Compared with the control group,the mRNA expression levels of STAT1 and C/EBPε,and the protein expression levels of STAT1 and p-STAT1 in the observation group at 48,72,and96 h after ATRA induction increased (all P ＜ 0.05),while the expression of miR-155-5p decreased (P ＜ 0.05).Pearson correlation analysis showed that STAT1 mRNA was negatively correlated with miR-155-5p (r =-0.90,P ＜0.05),while was positively correlated with C/EBPε mRNA (r =0.96,P ＜ 0.05).Conclusions The miR-155-5p reversely regulates STAT1 signaling pathway,thus activates the transcription of C/EBPε,which leads to the differentiation of HL-60 cells into granulocytes.Activation of STAT1 signaling pathway may be one of the key mechanisms in the differentiation of HL-60 cells into granulocytes,and STAT1 may be taken as a novel target for clinical treatment.
Keywords：human acute myeloid leukemia cellsHL-60 cellsgranulocytesall-trans retinoic acidsignal transducer and activator of transcription 1microRNA-155-5pCCAAT/enhancer binding proteins ε
Publication Date：2018-01-01
Online Publishing Date：2025-08-15(First online date of this platform, not the publication date of the document)
Pages：4( 24-27 )