Upregulation and functional roles of miR-450b in canine oral melanoma

Md Nazmul Hasan; Md Mahfuzur Rahman; Al Asmaul Husna; Mohammad Arif; Indre Jasineviciute; Daiki Kato; Takayuki Nakagawa; Naoki Miura

doi:10.1016/j.ncrna.2024.01.017

Upregulation and functional roles of miR-450b in canine oral melanoma

Noncoding RNA Res. 2024 Feb 1;9(2):376-387. doi: 10.1016/j.ncrna.2024.01.017. eCollection 2024 Jun.

Authors

Md Nazmul Hasan^{1

2}, Md Mahfuzur Rahman³, Al Asmaul Husna², Mohammad Arif¹, Indre Jasineviciute⁴, Daiki Kato⁵, Takayuki Nakagawa⁵, Naoki Miura^{1

2}

Affiliations

¹ Joint Graduate School of Veterinary Medicine, Kagoshima University, 1-21-24, Korimoto, Kagoshima, 890-0065, Japan.
² Veterinary Teaching Hospital, Joint Faculty of Veterinary Medicine, Kagoshima University, 1-21-24, Korimoto, Kagoshima, 890-0065, Japan.
³ Department of Human Oncology, University of Wisconsin School of Medicine and Public Health, Madison, WI, USA.
⁴ Department of Anatomy and Physiology, Veterinary Faculty, Lithuanian University of Health Sciences, LT-47181, Kaunas, Lithuania.
⁵ Laboratory of Veterinary Surgery, Graduate School of Agricultural and Life Sciences, The University of Tokyo, Tokyo, 113-8657, Japan.

Abstract

Canine oral melanoma (COM) is a common and highly aggressive disease with the potential to model human melanomas. Dysregulated microRNAs represent an interesting line of research for COM because they are implicated in tumor progression. One example is miR-450b, which has been investigated for its molecular mechanisms and biological functions in multiple human cancers, but not human or canine melanoma. Here, we aimed to investigate miR-450b as a potential diagnostic biomarker of COM and its functional roles in metastatic and non-metastatic forms of the disease. We investigated the expression of miR-450b and its target mRNA genes in clinical (tumor tissue and plasma) samples and metastatic and primary-tumor cell lines. Knockdown and overexpression experiments were performed to determine the influence of miR-450b on cell proliferation, migration, colony formation, and apoptosis. miR-450b was significantly upregulated in COM and differentiated between metastatic and non-metastatic tumors, and its potential as a biomarker of metastatic and non-metastatic COM was further confirmed in ROC analysis. miR-450b knockdown promoted cell proliferation, migration, and clonogenicity and inhibited apoptosis, whereas its overexpression yielded the reverse pattern. miR-450b directly binds 3' UTR of PAX9 mRNA and modulates its function leading to BMP4 downregulation and MMP9 upregulation at the transcript level. Furthermore, we surmised that miR-450b activates the Wnt signaling pathway based on gene ontology and enrichment analyses. We concluded that miR-450b has the potential as a diagnostic biomarker and could be a target candidate for COM treatment.

Keywords: Apoptosis; Colony formation; Migration; PAX9-BMP4-MMP9; Proliferation; miR-450b.