Nabeta Rina, Kanaya Ami, Shimada Kazumi, Matsuura Katsuhiro, Yoshimura Aritada, Oyamada Tomohiro, Azakami Daigo, Furuya Tetsuya, Uchide Tsuyoshi
Laboratory of Veterinary Molecular Pathology and Therapeutics, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan.
Laboratory of Veterinary Surgery, Faculty of Agriculture, Tokyo University of Agriculture and Technology, Fuchu, Japan.
Front Vet Sci. 2024 Sep 9;11:1436621. doi: 10.3389/fvets.2024.1436621. eCollection 2024.
Canine mesotheliomas are uncommon malignant tumors typically detected late. Minimally invasive diagnostic biomarkers would facilitate diagnosis at earlier stages, thereby improving clinical outcomes. We hypothesized that mesothelin could be used as a reliable diagnostic biomarker for canine mesotheliomas since it has been used as a cancer biomarker for human mesothelioma. We aimed to explore and characterize mesothelin gene expression in dogs and assess its use as a diagnostic biomarker for canine mesotheliomas.
We quantified expressed canine mesothelin transcripts via reverse transcription polymerase chain reaction (RT-PCR) and sequenced them using ribonucleic acid (RNA) extracted from a canine mesothelioma cell line. After confirming mesothelin expression, we assessed its levels in major organ tissues and compared them with those in the mesothelioma tissues using quantitative PCR (qPCR). Mesothelin overexpression in mesotheliomas was detected, and we further compared its levels using qPCR between mesotheliomas and non-mesotheliomas using tumor tissues and clinical sample effusions, confirming its significance as a diagnostic biomarker for canine mesothelioma.
Mesothelin complementary deoxyribonucleic acid (cDNA) was amplified via RT-PCR, yielding a single band of expected upon DNA electrophoresis. Sequence analyses confirmed it as a predicted canine mesothelin transcript from the genome sequence database. Comparative sequence analysis of the deduced amino acid sequence of the expressed canine mesothelin demonstrated molecular signature similarities with the human mesothelin. However, the pre-sequence of canine mesothelin lacks the mature megakaryocyte potentiating factor (MPF) portion, which is typically cleaved post-translationally with furin. Mesothelin expression was quantified via qPCR revealing low levels in the mesothelial and lung tissues, with negligible expression in the other major organs. Canine mesothelin exhibited significantly higher expression in the canine mesotheliomas than in the noncancerous tissues. Moreover, analysis of clinical samples using qPCR demonstrated markedly elevated mesothelin expression in canine mesotheliomas compared to non-mesothelioma cases.
Canine mesothelin exhibits molecular and biological characteristics akin to human mesothelin. It could serve as a vital biomarker for diagnosing canine mesotheliomas, applicable to both tissue- and effusion-based samples.
犬类间皮瘤是一种罕见的恶性肿瘤，通常在晚期才被发现。微创诊断生物标志物将有助于早期诊断，从而改善临床预后。我们假设间皮素可作为犬类间皮瘤的可靠诊断生物标志物，因为它已被用作人类间皮瘤的癌症生物标志物。我们旨在探索和表征犬类间皮素基因在犬类中的表达，并评估其作为犬类间皮瘤诊断生物标志物的用途。
我们通过逆转录聚合酶链反应（RT-PCR）对表达的犬类间皮素转录本进行定量，并使用从犬类间皮瘤细胞系中提取的核糖核酸（RNA）对其进行测序。在确认间皮素表达后，我们使用定量PCR（qPCR）评估其在主要器官组织中的水平，并将其与间皮瘤组织中的水平进行比较。检测到间皮瘤中间皮素的过表达，我们进一步使用qPCR比较了间皮瘤与非间皮瘤之间的水平，使用肿瘤组织和临床样本积液，证实其作为犬类间皮瘤诊断生物标志物的意义。
通过RT-PCR扩增间皮素互补脱氧核糖核酸（cDNA），在DNA电泳时产生一条预期的单带。序列分析证实它是来自基因组序列数据库的预测犬类间皮素转录本。对表达的犬类间皮素推导氨基酸序列的比较序列分析表明与人类间皮素具有分子特征相似性。然而，犬类间皮素的前序列缺乏成熟的巨核细胞增强因子（MPF）部分，该部分通常在翻译后被弗林蛋白酶切割。通过qPCR对间皮素表达进行定量，结果显示间皮和肺组织中的水平较低，其他主要器官中的表达可忽略不计。犬类间皮素在犬类间皮瘤中的表达明显高于非癌组织。此外，使用qPCR对临床样本的分析表明，与非间皮瘤病例相比，犬类间皮瘤中间皮素的表达明显升高。
犬类间皮素表现出与人类间皮素相似的分子和生物学特征。它可作为诊断犬类间皮瘤的重要生物标志物，适用于基于组织和积液的样本。
