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Objective  To investigate the regulatory network of long non‐coding RNA (lncRNA) as competing endogenous RNAs (ceRNAs) in osteonecrosis of the femoral head (ONFH).    Methods  The gene expression profile GSE74089 of ONFH and microRNA (miRNA) expression profile of GSE89587 were obtained from the Gene Expression Omnibus (GEO) database. The GSE74089 contained four ONFH samples and four controls. The GSE89587 included 10 ONFH samples and 10 control samples. The differentially expressed lncRNAs (DE‐lncRNAs) and DE‐mRNAs between ONFH group and control group were identified from GSE74089 using the limma package based on criteria of adjusted  P  value <0.05 and |log fold change (FC)| ≥2. The DEmiRNAs between ONFH group and control group were screened from GSE89587 on the basis of adjusted  P  value <0.05. Gene ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway for DE‐mRNAs were analyzed using DAVID 6.7 and GSEA 3.0, respectively. Coexpressed lncRNA‐mRNA pairs were identified by corr.test method in R based on the criteria of adjusted  P  value <0.01 and |r| ≥ 0.9. A ceRNA network was constructed and visualized using cytoscape 3.7.0 by integrating the DE‐lncRNA, DE‐miRNA, and DEmRNA data. The key mRNAs and lncRNAs in the ceRNA network were further validated in an independent dataset of GSE123568.    Results  Based on our analysis, a total of 28 DE‐lncRNAs, 1403 DE‐mRNAs, and 134 DE‐miRNAs were identified, respectively. The DE‐mRNAs were significantly enriched in the function of “skeletal system development,” “collagen fibril organization,” “blood vessel development,” and “regulation of nervous system development.” Besides, 72 KEGG pathways, including eight active pathways and 64 suppressed pathways were identified, including which immune pathway was the most significantly activated one and which ribosome‐related function was the most suppressed. A co‐expression network including 161 DE‐mRNAs and 16 DE‐lncRNAs was built. Highly connected nodes were identified among lncRNAs such as  H19 ,  C20orf203 ,  LINC00355 ,  SFTA3 ,  CRNDE ,  CASC2 ,  LINC00494 ,  C9orf163 ,  C10orf91 , and  LINC00301 . The ceRNA network indicated that lncRNA  H19  functioned as a ceRNA of hsa‐miR‐519b‐3p and hsa‐miR‐296‐5p in  ANKH  and  ECHDC1  regulation; lncRNA  C9orf163  functioned as a ceRNA of hsa‐miR‐424‐5p in  CCNT1  regulation. The expression trends of  ANKH ,  CCNT1 , and  C9orf163  were successfully validated in independent dataset of GSE123568.    Conclusion  The ceRNAs of lncRNA  H19 ‐ hsa‐miR‐519b‐3p/hsa‐miR‐296‐5p‐ ANKH  and lncRNA  c9orf163‐  hsa‐miR‐424‐5p‐ CCNT1  might play important roles in ONFH development. Our research provided an understanding of the important role of lncRNA–related ceRNAs in ONFH.

Non‐coding RNA Identification in Osteonecrosis of the Femoral Head Using Competitive Endogenous RNA Network Analysis