实用医学杂志2025，Vol.41Issue(4)：478-489,12.DOI:10.3969/j.issn.1006-5725.2025.04.004
环状RNA mmu_circ_0000818在地塞米松导致的MC3T3-E1细胞凋亡中的作用及机制
Role and mechanism of circular RNA mmu_circ_0000818 in dexamethasone-induced apoptosis of MC3T3-E1 cells
摘要
Abstract
Objective To screen for differentially expressed apoptosis-related circular RNAs(circRNAs)in osteoblasts from steroid-induced osteonecrosis of the femoral head(SONFH)and to investigate their roles and mechanisms in osteoblast apoptosis.Methods MC3T3-E1 cells were cultured and divided into two groups:Control and DEX-treated.Western blot analysis was employed to evaluate the expression levels of BCL2-Associated X protein(Bax)and B-cell lymphoma 2(Bcl-2).Cell apoptosis was assessed using TUNEL staining and flow cytometry.RNA was extracted from both normal and DEX-treated MC3T3-E1 cells,followed by RNA-seq to identify differen-tially expressed circular RNAs(circRNAs).The functions and pathways of these circRNAs were analyzed using Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG).The differentially expressed mmu_circ_0000818 was selected for further verification at the cellular level.Its chromosomal location and conservation were examined using the UCSC Genome Browser Gateway and circBase.Overexpression plasmids and small interfering RNAs(siRNAs)targeting mmu_circ_0000818 were constructed and transfected into the cells.Subsequently,apop-tosis in MC3T3-E1 cells from each group was evaluated by flow cytometry.Results Compared with the control group,the apoptosis rate of MC3T3-E1 cells was significantly increased in the DEX group(P<0.01).Differen-tially expressed circRNAs were identified based on log2foldchange(≥2)and P value(P<0.05).Relative to the control group,there were 234 differentially expressed circRNAs in the DEX group,including 138 up-regulated and 96 down-regulated circRNAs.GO and KEGG enrichment analyses of the target genes of these differentially expressed circRNAs revealed significant associations with apoptosis and the PI3K-Akt signaling pathway.qRT-PCR results demonstrated that the expression level of mmu_circ_0000818 was markedly higher in the DEX group com-pared to the control group(P<0.01).Analysis using the UCSC Genome Browser and CircBase indicated that mmu_circ_0000818,located at chromosome 17:78712463-78715086,is formed by the cyclization of exons 6-7 of the Crim1 gene and exhibits high conservation across species.Flow cytometry results indicated that knockdown of mmu_circ_0000818 attenuated DEX-induced apoptosis in MC3T3-E1 cells,while overexpression of mmu_circ_0000818 exacerbated apoptosis.Conclusions CircRNA mmu_circ_0000818 was significantly upregulated in DEX-treated MC3T3-E1 cells,and its downregulation mitigated DEX-induced apoptosis.Consequently,mmu_circ_0000818 may represent a promising therapeutic target for the prevention and treatment of SONFH.关键词
环状RNA mmu_circ_0000818/地塞米松/MC3T3-E1细胞/RNA测序/凋亡Key words
circRNA mmu_circ_0000818/dexamethasone/MC3T3-E1 cells/RNA sequencing/apoptosis分类
中医学引用本文复制引用
杨慧霞,丁宁,马润秋,李桂忠,郝银菊,马胜超,姜怡邓,白志刚..环状RNA mmu_circ_0000818在地塞米松导致的MC3T3-E1细胞凋亡中的作用及机制[J].实用医学杂志,2025,41(4):478-489,12.基金项目
国家自然科学基金项目(编号:82060412) （编号:82060412）
国家自然科学基金区域创新发展联合基金项目(编号:U21A20343) （编号:U21A20343）
宁夏回族自治区重点研发计划项目(编号:2020BFH02001,2022BEG02054,2022BFH02013,2023BEG02074) （编号:2020BFH02001,2022BEG02054,2022BFH02013,2023BEG02074）
