MicroRNA-655-3p and microRNA-497-5p inhibit cell proliferation in cultured human lip cells through the regulation of genes related to human cleft lip

Abstract

Background The etiology of cleft lip with or without cleft palate (CL/P), a common congenital birth defect, is complex and involves the contribution of genetic and environmental factors. Although many candidate genes have been identified, the regulation and interaction of these genes in CL/P remain unclear. In addition, the contribution of microRNAs (miRNAs), non-coding RNAs that regulate the expression of multiple genes, to the etiology of CL/P is largely unknown. Methods To identify the signatures of causative biological pathways for human CL/P, we conducted a systematic literature review for human CL/P candidate genes and subsequent bioinformatics analyses. Functional enrichment analyses of the candidate CL/P genes were conducted using the pathway databases GO and KEGG. The miRNA-mediated post-transcriptional regulation of the CL/P candidate genes was analyzed with miRanda, PITA, and TargetScan, and miRTarbase. Genotype-phenotype association analysis was conducted using GWAS. The functional significance of the candidate miRNAs was evaluated experimentally in cell proliferation and target gene regulation assays in human lip fibroblasts. Results Through an extensive search of the main biomedical databases, we mined 177 genes with mutations or association/linkage reported in individuals with CL/P, and considered them as candidate genes for human CL/P. The genotype-phenotype association study revealed that mutations in 12 genes ( ABCA4 , ADAM3A , FOXE1 , IRF6 , MSX2 , MTHFR , NTN1 , PAX7 , TP63 , TPM1 , VAX1 , and WNT9B ) were significantly associated with CL/P. In addition, our bioinformatics analysis predicted 16 microRNAs (miRNAs) to be post-transcriptional regulators of CL/P genes. To validate the bioinformatics results, the top six candidate miRNAs (miR-124-3p, miR-369-3p, miR-374a-5p, miR-374b-5p, miR-497-5p, and miR-655-3p) were evaluated by cell proliferation/survival assays and miRNA-gene regulation assays in cultured human lip fibroblasts. We found that miR-497-5p and miR-655-3p significantly suppressed cell proliferation in these cells. Furthermore, the expression of the predicted miRNA-target genes was significantly downregulated by either miR-497-5p or miR-655-3p mimic. Conclusion Expression of miR-497-5p and miR-655-3p suppresses cell proliferation through the regulation of human CL/P-candidate genes. This study provides insights into the role of miRNAs in the etiology of CL/P and suggests possible strategies for the diagnosis of CL/P. Electronic supplementary material The online version of this article (10.1186/s12920-019-0535-2) contains supplementar...

LINC00473 contributes to the radioresistance of esophageal squamous cell carcinoma by regulating microRNA‑497‑5p and cell division cycle 25A

“…miRs have thus been shown to play pivotal roles in the biological functions of cancer cells, such as differentiation, the stress response, proliferation, apoptosis and sensitivity to irradiation (19). Various miRs of the miR-15/16 family, including miR-15a-3p, miR-16-1-3p and miR-497, exhibit anticancer behaviors (20)(21)(22)(23)(24)(25)(26). For instance, miR-497 has been shown to suppress the malignant behavior of papillary thyroid cancer cells by reversely modulating the expression of Yes associated protein 1 (YAP1) (22); it has also been shown to inhibit the angiogenesis and metastasis of hepatocellular carcinoma by impeding the actions of vascular endothelial growth factor A and astrocyte elevated gene-1 (23).…”

Role of epigenetics and miRNAs in orofacial clefts

“…Additionally, among other targets, miR‐374a‐5p was validated to target MSX1 , while miR‐4680‐3p was validated to target ERBB2 and an enzyme involved in folate metabolism, MTHFD1 . Gajera et al (2019) utilized a similar approach using data from CL/P patients (Gajera et al, 2019). miR‐497‐5p and miR‐655‐3p mimics were able to suppress proliferation in human lip cell cultures.…”

lncRNA SNHG5 promotes cell proliferation, migration and invasion in oral squamous cell carcinoma by sponging miR‑655‑3p/FZD4 axis

“…Downregulation of miR-655-3p has been detected in retinoblastoma and esophageal squamous cell carcinoma ( 12 , 13 ). Functionally, miR-655-3p was found to inhibit cell proliferation in human lip cells ( 14 ). In addition, miR-655-3p restrained the migration and invasion of non-small cell lung cancer cells by targeting PTTG1 ( 15 ).…”