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CircSSRP1 regulates SMAD3 to affect proliferation and apoptosis of human embryonic palatal mesenchymal cells in nonsyndromic cleft lip with or without cleft palate by sponging miR-708-5p

ABSTRACT: Nonsyndromic cleft lip with or without cleft palate (NSCL/P) represents one of the most prevalent congenital anomalies in oral and maxillofacial region with unclear etiology. It is currently widely accepted that NSCL/P is a complex genetic disorder caused by multiple environmental and genetic factors. This study aimed to investigate the regulatory roles of circSSRP1/miR-708-5p/SMAD3 axis in NSCL/P. Using bioinformatic analysis methods, we performed an integrated analysis of the Clariom D Human Assay and GSE42589 dataset to construct the NSCL/P associated circSSRP1/miR-708-5p/SMAD3 axis. Subsequently, through validating the expressions of circSSRP1, miR-708-5p, and SMAD3 in NSCL/P tissues and matched normal control tissues, combined with in vitro experiments and other relevant approaches, we identified a novel circular RNA, circSSRP1. This molecule acts as a molecular sponge for miR-708-5p, thereby regulating the expression of SMAD3 and participating in the pathogenesis of NSCL/P.

ORGANISM(S): Homo sapiens

PROVIDER: GSE316908 | GEO | 2026/07/05

REPOSITORIES: GEO

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Project description:The etiology and pathogenesis of non-syndromic cleft lip and palate (NSCL/P) are largely unknown. Long non-coding RNAs (lncRNA) are thought to play important roles in NSCL/P, but reports on the underlying processes are currently unavailable. Our study focused on children diagnosed with NSCL/P alone. Based on the morphology, patients were categorized as either cleft lip with or without cleft palate (CL/P) or cleft palate-only (CPO). When patients received surgery for NSCL/P, tissue excised from the trimmed wound edge was reserved to serve as experimental samples; adjacent normal tissue was used as a positive control. Target lncRNAs in the collected tissues were identified using microarray and quantitative reverse transcription PCR (RT-qPCR). Immunohistochemical (IHC) staining and RT-qPCR were used to verify the target mRNAs. Pathway, gene ontology (GO) enrichment, and TargetScan prediction were employed to construct endogenous RNA networks (ceRNA networks) and explore their potential functions. RNA-Seq analysis revealed 24 upregulated and 43 downregulated lncRNAs in the CL/P and CPO groups compared with those in the control group; of these, MALAT1and NEAT1 were screened and validated using RT-qPCR. Common NSCL/P risk factors positively correlated with MALAT1 and NEAT1 expression (ORMALAT1 = 28.111, 95% CI: 4.054-194.923; ORNEAT1 = 30.556, 95% CI: 4.422-211.142; P < 0.05). Bioinformatics predicted four ceRNA networks: MALAT1-hsa-miR-1224-3p-SP1, MALAT1-hsa-miR-6734-5p/hsa-miR-1224-3p-WNT10A, NEAT1-hsa-miR-140-3p.1-CXCR4, and NEAT1-hsa-miR-3129-5p/hsa-miR-199a-3p/hsa-miR-199b-3p-ZEB1. GO enrichment focused on the potential functions of ceRNA networks, including biosynthesis of organic cyclic compounds, formation of membrane-enclosed and organelle lumens, and Wnt-protein binding. The results of RT-qPCR were consistent with those of IHC staining with regard to expression of related mRNAs. MALAT1 and NEAT1, which are upregulated in NSCL/P, are associated with the severity of NSCL/P. This study provides a new insight into NSCL/P pathogenesis and suggests that MALAT1 and NEAT1 act as potential therapeutic targets and prognostic biomarkers for NSCL/P.

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CircSSRP1 regulates SMAD3 to affect proliferation and apoptosis of human embryonic palatal mesenchymal cells in nonsyndromic cleft lip with or without cleft palate by sponging miR-708-5p

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