Phenytoin Inhibits Cell Proliferation through microRNA-196a-5p in Mouse Lip Mesenchymal Cells

• Published in: International journal of molecular sciences 2021-02, Vol.22 (4), p.1746
• Main Authors: Yoshioka, Hiroki, Ramakrishnan, Sai Shankar, Suzuki, Akiko, Iwata, Junichi
• Format: Article
• Language: English
• Subjects: Animals; Binding sites; Birth defects; Cell growth; Cell Line; Cell proliferation; Cell Proliferation - drug effects; Cell Proliferation - genetics; Chromosome 5; cleft lip; Cleft Lip - chemically induced; Cleft Lip - genetics; Cleft Lip - pathology; Cleft lip/palate; Congenital defects; craniofacial development; Disease Models, Animal; Embryo, Mammalian; environmental factor; Female; Gene expression; Gene Expression Regulation, Developmental - drug effects; gene regulation; Genes; Humans; Lip - cytology; Lip - embryology; Maternal Exposure - adverse effects; Mesenchymal Stem Cells - drug effects; Mesenchyme; Mice; microRNA; MicroRNAs; MicroRNAs - antagonists & inhibitors; MicroRNAs - metabolism; miRNA; Mouse Embryonic Stem Cells; Mutation; Neural crest; Phenytoin; Phenytoin - toxicity; Primary Cell Culture; Teratogens - toxicity
• ISSN: 1422-0067; 1661-6596; 1422-0067
• DOI: 10.3390/ijms22041746

Cleft lip (CL) is one of the most common birth defects. It is caused by either genetic mutations or environmental factors. Recent studies suggest that environmental factors influence the expression of noncoding RNAs [e.g., microRNA (miRNA)], which can regulate the expression of genes crucial for cellular functions. In this study, we examined which miRNAs are associated with CL. Among 10 candidate miRNAs (miR-98-3p, miR-101a-3p, miR-101b-3p, miR-141-3p, miR-144-3p, miR-181a-5p, miR-196a-5p, miR-196b-5p, miR-200a-3p, and miR-710) identified through our bioinformatic analysis of CL-associated genes, overexpression of miR-181a-5p, miR-196a-5p, miR-196b-5p, and miR-710 inhibited cell proliferation through suppression of genes associated with CL in cultured mouse embryonic lip mesenchymal cells (MELM cells) and O9-1 cells, a mouse cranial neural crest cell line. In addition, we found that phenytoin, an inducer of CL, decreased cell proliferation through miR-196a-5p induction. Notably, treatment with a specific inhibitor for miR-196a-5p restored cell proliferation through normalization of expression of CL-associated genes in the cells treated with phenytoin. Taken together, our results suggest that phenytoin induces CL through miR-196a-5p induction, which suppresses the expression of CL-associated genes.