Elevated expression of miR-301a and its functional roles in canine oral melanoma.

IF 2.3 2区 农林科学 Q1 VETERINARY SCIENCES Veterinary and comparative oncology Pub Date : 2024-03-01 Epub Date: 2023-12-26 DOI:10.1111/vco.12954
Md Nazmul Hasan, Md Mahfuzur Rahman, Al Asmaul Husna, Mohammad Arif, Tomoko Iwanaga, Kyoko Tsukiyama-Kohara, Indre Jasineviciute, Daiki Kato, Takayuki Nakagawa, Naoki Miura
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Abstract

miR-301a is one of numerous dysregulated microRNAs (miRNAs) in canine oral melanoma (COM), one of which is miR-301a (upregulated). Its biological role has been described in various human cancer types, including malignant melanoma, but not in COM. Accordingly, in this study, we investigated miR-301a expression in COM in greater detail to ascertain whether it could serve as a diagnostic biomarker, elucidate its functional roles in this cancer, and predict the possible pathways by which it exerts its effects. Relative expression of miR-301a was investigated in clinical oral tissue and plasma samples and COM cell (KMeC and LMeC) lines using qRT-PCR. Knockdown of miR-301a was also validated for KMeC and LMeC cells using qRT-PCR. We performed CCK-8 assays to assess cell proliferation, monolayer wound-healing, and transwell migration assays to assess cell migration, a colony-formation assay to assess clonogenicity, a TUNEL assay and flow cytometry to assess apoptosis-related effects, and gene enrichment analyses to predict possible related pathways. miR-301a was markedly upregulated in COM oral tissue and plasma clinically, suggesting its potential as a diagnostic biomarker for COM diagnosis. In vitro assays demonstrated that miR-301 significantly inhibited apoptosis in COM cells while promoting cell migration, proliferation, and clonogenicity. We also predicted that miR-301 exerts cancer-promoting effects through the Wnt signalling pathway for COM. Our findings suggest that miR-301a is a COM oncomiR that regulates several oncogenic phenotypes with the potential to be a diagnostic biomarker.

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犬口腔黑色素瘤中 miR-301a 的高表达及其功能作用。
miR-301a是犬口腔黑色素瘤(COM)中众多失调的微RNA(miRNA)之一,其中之一是miR-301a(上调)。其生物学作用已在包括恶性黑色素瘤在内的多种人类癌症类型中得到描述,但在 COM 中却没有。因此,在本研究中,我们更详细地调查了 miR-301a 在 COM 中的表达情况,以确定它是否可作为诊断生物标志物,阐明它在这种癌症中的功能作用,并预测其发挥效应的可能途径。利用 qRT-PCR 技术研究了临床口腔组织和血浆样本以及 COM 细胞系(KMeC 和 LMeC)中 miR-301a 的相对表达。还利用 qRT-PCR 验证了 KMeC 和 LMeC 细胞中 miR-301a 的敲除。我们进行了 CCK-8 试验以评估细胞增殖,单层伤口愈合和跨孔迁移试验以评估细胞迁移,集落形成试验以评估克隆性,TUNEL 试验和流式细胞术以评估细胞凋亡相关效应,以及基因富集分析以预测可能的相关通路。体外实验表明,miR-301 能显著抑制 COM 细胞的凋亡,同时促进细胞迁移、增殖和克隆。我们还预测,miR-301 通过 Wnt 信号通路对 COM 发挥促癌作用。我们的研究结果表明,miR-301a 是一种调控多种致癌表型的 COM oncomiR,有望成为一种诊断生物标志物。
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来源期刊
Veterinary and comparative oncology
Veterinary and comparative oncology 农林科学-兽医学
CiteScore
4.80
自引率
9.50%
发文量
75
审稿时长
>24 weeks
期刊介绍: Veterinary and Comparative Oncology (VCO) is an international, peer-reviewed journal integrating clinical and scientific information from a variety of related disciplines and from worldwide sources for all veterinary oncologists and cancer researchers concerned with aetiology, diagnosis and clinical course of cancer in domestic animals and its prevention. With the ultimate aim of diminishing suffering from cancer, the journal supports the transfer of knowledge in all aspects of veterinary oncology, from the application of new laboratory technology to cancer prevention, early detection, diagnosis and therapy. In addition to original articles, the journal publishes solicited editorials, review articles, commentary, correspondence and abstracts from the published literature. Accordingly, studies describing laboratory work performed exclusively in purpose-bred domestic animals (e.g. dogs, cats, horses) will not be considered.
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