CircRNA screening and ceRNA network construction for key regulation of weightlessness-induced bone loss

WANG Dong; LI Chengfei; LI Xi; ZHAO Ziyi; LI Jiaxiang; YAN Ming; WANG Yongchun

doi:10.13276/j.issn.2097-1656.2025.01.004

Abstract：Objective To screen differential expression profiles of circular RNA(circRNA)in bone marrow mesenchymal stem cells(BMSCs)under simulated weightlessness by high-throughput sequencing(HTS)technology,conduct bioinformatics analysis and partial verification of the differentially expressed circRNA(DEcircRNA)to explore the possible mechanisms of weightlessness-induced bone loss at the molecular level.Methods Osteogenic differentiation of mouse BMSCs was induced by bone morphogenetic protein-2 and divided into Ground group and simulated microgravity group(SMG group).The total RNA of the samples was sequenced by Illumina NovaSeq 6000,and differentially expressed genes(DEGs)of the two groups were screened and analyzed to further construct the competing endogenous RNA(ceRNA)regulatory network of circRNA-miRNA-mRNA interactions.Results A total of 18 DEcircRNAs were identified by comparison between the two groups,including 7 up-regulated and 11 down-regulated circRNAs.GO and KEGG enrichment analysis showed that the source genes of these DEcircRNAs were mainly enriched in the functions of regulating cell differentiation and cell proliferation,and were significantly enriched in MAPK signaling pathway and cell aging signaling pathway.The establishment of ceRNA regulatory network could provide a better understanding of the biological role of DEcircRNA.Six DEcircRNAs were verified by qRT-PCR,and the results were consistent with the sequencing results.Conclusion This study screens and clarifies the potential biological role of 18 DEcircRNAs in weightlessness-induced bone loss through HTS and bioinformatics methods,providing a new idea for the subsequent study of the mechanism and clinical treatment of weightlessness-induced bone loss.