Transcriptional regulation of interleukin-6 receptor by PML::RARα fusion protein

Abstract

Abstract:

ObjectiveTo investigate the regulatory mechanism of PML::RARα fusion protein on interleukin-6 receptor (IL-6R) and the effects of IL-6R on the proliferation and differentiation of acute promyelocytic leukemia (APL) cells.MethodsThe expression levels of IL-6R in APL cells were analyzed using the GSE12662 and GSE10358 datasets. Reverse transcription quantitative real-time quantitative PCR（RT-qPCR） was performed to detect IL-6R mRNA expression in NB4 cells before and after all-trans retinoic acid (ATRA) treatment, as well as in PR9 cells before and after Zn²⁺ induction. Chromatin immunoprecipitation (ChIP)-seq data analysis, ChIP-qPCR experiments, and luciferase reporter gene activity assays were performed to explore the regulatory mechanism of PML::RARα on IL-6R. An IL-6R expression plasmid was constructed for NB4 cells via retrovirus. Cell proliferation was assessed using the cell counting kit-8 (CCK-8) assay, and CD11b expression was detected by flow cytometry.ResultsAnalysis of the GSE12662 dataset revealed that the expression level of IL-6R in APL cells (12.20 ± 0.41) was significantly lower than that in normal promyelocytes (13.14 ± 0.47,t= 4.289,P< 0.001) and polymorphonuclear cells (14.82 ± 0.40,t= 12.35,P< 0.001). Moreover, analysis of the GSE10358 dataset showed that IL-6R expression in APL patients (5.93 ± 0.84) was significantly lower than that in non-APL AML patients (6.50 ± 0.87,t= 3.91,P< 0.001). PML::RARα directly bound to the promoter region of IL-6R to inhibit its transcriptional activity resulting in the low expression. Overexpression of IL-6R in the APL-derived NB4 cells significantly inhibited cell proliferation. Four days after transfection, the optical density values measured by the CCK-8 assay were 0.86 ± 0.01 and 0.40 ± 0.01, respectively (t= 32.66,P< 0.001). Simultaneously, cell differentiation was significantly enhanced. The ratio of the CD11b positive cells increased from 3.10 % ± 1.22 % to 14.4 % ± 1.11 % (t= 11.84,P< 0.001).ConclusionsIL-6R is a target gene of PML::RARα, demonstrating that PML::RARα can suppress IL-6R transcription by binding to its promoter region. It is illustrated that IL-6R inhibited the cell proliferation and induced partial differentiation in APL cells.

Key words:Interleukin-6 receptor,Acute promyelocytic leukemia,PML::RARα,Cell proliferation,Cell differentiation

CLC Number:

R733.7

ZHAO Lingling, CUI Canqi, MI Jianqing. Transcriptional regulation of interleukin-6 receptor by PML::RARα fusion protein[J]. Journal of Internal Medicine Concepts & Practice, 2025, 20(02): 146-151.

Figures/Tables3

转染天数（d）	空载	IL-6R	t	P
0	0.24 ± 0.00	0.24 ± 0.00	0.13	0.90
1	0.34 ± 0.02	0.31 ± 0.01	2.22	0.09
2	0.39 ± 0.01	0.34 ± 0.02	4.01	0.02
3	0.54 ± 0.01	0.35 ± 0.01	35.74	<0.0001
4	0.86 ± 0.01	0.40 ± 0.01	32.66	<0.0001

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