The role and mechanism of LncRNA GABPB1-AS1 in regulating proliferation,invasion,migration,and apoptosis of acute myeloid leukemia cells
10.12354/j.issn.1000-8179.2024.20240164
- VernacularTitle:LncRNA GABPB1-AS1调控急性髓系白血病细胞增殖 侵袭迁移及凋亡的作用和机制
- Author:
Yang FEIFEI
1
;
Tang HAO
;
Sun HUI
;
Sun YIN
Author Information
1. 滕州市中心人民医院儿科(山东省滕州市 277500)
- Keywords:
acute myeloid leukemia(AML);
antisense RNA of GA binding protein transcription factor β subunit 1(GABPB1-AS1);
prolif-eration;
invasion;
apoptosis
- From:
Chinese Journal of Clinical Oncology
2024;51(8):392-400
- CountryChina
- Language:Chinese
-
Abstract:
Objective:To explored the role and mechanism of antisense RNA of long non-coding RNA(LncRNA)GA binding protein transcrip-tion factor β subunit 1(GABPB1-AS1)in regulating the proliferation,invasion,migration,and apoptosis of acute myeloid leukemia(AML)cells by targeting the microRNA-497-5p/heat shock protein 8(miR-497-5p/HSPA8)axis.Methods:HL-60 cells were divided into the normal con-trol,si-NC,si-GABPB1-AS1,si-GABPB1-AS1+NC,and si-GABPB1-AS1+miR-497-5p inhibitor groups.Quantitative real time polymerase chain re-action(RT-qPCR)was used to detect the expression levels of LncRNA GABPB1-AS1,miR-497-5p,and HSPA8.Double luciferase reporter gene assay was used to verify the targeting relationship between LncRNA GABPB1-AS1,miR-497-5p,and HSPA8;MTT was used to detect cell viab-ility;while 5-ethynyl-2'-deoxyuridine(EdU)was used to detect proliferation.Transwell chamber experiments were used to detect invasion and migration while flow cytometry was used to detect apoptosis.Western blot was used to detect the levels of proliferating cell nuclear an-tigen(PCNA),HSPA8,metastasis-associated proteins(MTA2),homolog of yeast Atg6(Beclin-1),and Caspase-3 proteins.A mouse trans-planted tumor model was established to verify the effect of LncRNA GABPB1-AS1 on the growth of AML transplanted tumors.Results:Com-pared to human bone marrow monocytes,LncRNA GABPB1-AS1 was highly expressed[(1.29±0.10),(1.58±0.12),(2.02±0.17),(3.17±0.24)vs.(1.02±0.07)]while miR-497-5p was lowly expressed[(0.94±0.07),(0.75±0.03),(0.57±0.03),(0.25±0.01)vs.(1.05±0.09)]in different AML cells(THP-1,NB4,U-937,and HL-60,respectively).The HL-60 cell line was selected for functional verification experiments since LncRNA GABPB1-AS1 expression was highest in the HL-60 cells.Knockdown of LncRNA GABPB1-AS1 reduced HL-60 cell viability,the EdU positive rate,cell in-vasion and migration,the expression of HSPA8 mRNA,and HSPA8,PCNA,and MTA2 protein contents.It increased the apoptosis rate,and the expression of miR-497-5p,Caspase-3 and Beclin-1 protein(P<0.05).miR-497-5p had a targeting relationship with LncRNA GABPB1-AS1 and HSPA8;inhibiting the expression of miR-497-5p reversed the inhibitory effect of LncRNA GABPB1-AS1 knockdown on the malignant bio-logical behavior of HL-60 cells.Meanwhile,inhibiting the expression of LncRNA GABPB1-AS1 constrained the growth of transplanted tumors.Conclusions:Knockdown of LncRNA GABPB1-AS1 inhibits the progression of AML cells,which may be related to the upregulation of miR-497-5p expression and downregulation of HSPA8 expression.
