Okabayashi Ken, Narita Takanori, Takashiro Saki, Nadaoka Sawako, Kanai Shuichiro, Ito Daisuke, Kitagawa Masato
Department of Veterinary Medicine, College of Bioresource Sciences, Nihon University, 1866 Kameino, Fujisawa, Kanagawa 252-0880, Japan.
J Vet Med Sci. 2019 Mar 14;81(3):369-372. doi: 10.1292/jvms.18-0491. Epub 2019 Jan 22.
This study was undertaken to establish a method for measuring mRNA expression by using real-time RT-PCR in the diagnosis of canine meningiomas. When performing real-time RT-PCR, it is essential to include appropriate control tissues and to select appropriate housekeeping genes as an internal standard. Based on the results of our study, RPS18 constitutes a suitable internal standard for the comparison of mRNA expression between normal meninges and meningiomas. The results showed increased mRNA expression of VEGFA and EGFR; however, mRNA expression of KDR was reduced. Measuring mRNA expression by using real-time RT-PCR with appropriate control tissues and internal standards can provide useful information to understanding the pathogenesis of canine meningiomas, which corresponds with immunohistochemical findings.
本研究旨在建立一种利用实时逆转录聚合酶链反应(real-time RT-PCR)测量mRNA表达的方法,用于犬脑膜瘤的诊断。进行实时RT-PCR时,纳入适当的对照组织并选择合适的管家基因作为内标至关重要。根据我们的研究结果,核糖体蛋白S18(RPS18)构成了用于比较正常脑膜和脑膜瘤之间mRNA表达的合适内标。结果显示,血管内皮生长因子A(VEGFA)和表皮生长因子受体(EGFR)的mRNA表达增加;然而,激酶插入域受体(KDR)的mRNA表达降低。使用实时RT-PCR并结合适当的对照组织和内标来测量mRNA表达,可为理解犬脑膜瘤的发病机制提供有用信息,这与免疫组化结果相符。