Yokoyama Minako, Fujita Toshitsugu, Kadonosawa Yuka, Tatara Yota, Motooka Daisuke, Ikawa Masahito, Fujii Hodaka, Yokoayama Yoshihito
Department of Obstetrics and Gynecology, Graduate School of Medicine, Hirosaki University, 5 Zaifu-cho, 036-8562, Hirosaki, Aomori, Japan.
Department of Biochemistry and Genome Biology, Graduate School of Medicine, Hirosaki University, 5 Zaifu-cho, 036-8562, Hirosaki, Aomori, Japan.
Mol Biol Rep. 2023 Jan;50(1):531-540. doi: 10.1007/s11033-022-07994-x. Epub 2022 Nov 9.
Carbonyl reductase 1 (CBR1) is a nicotinamide adenine dinucleotide phosphate (NADPH)-dependent reductase with broad substrate specificity. CBR1 catalyzes the reduction of numerous carbonyl compounds, including quinones, prostaglandins, menadione, and multiple xenobiotics, while also participating in various cellular processes, such as carcinogenesis, apoptosis, signal transduction, and drug resistance. In this study, we aimed to generate transgenic mice overexpressing mouse Cbr1 (mCbr1), characterize the mCbr1 expression in different organs, and identify changes in protein expression patterns.
To facilitate a deeper understanding of the functions of CBR1, we generated transgenic mice overexpressing CBR1 throughout the body. These transgenic mice overexpress 3xFLAG-tagged mCbr1 (3xFLAG-mCbr1) under the CAG promoter. Two lines of transgenic mice were generated, one with 3xFLAG-mCbr1 expression in multiple tissues, and the other, with specific expression of 3xFLAG-mCbr1 in the heart. Pathway and network analysis using transgenic mouse hearts identified 73 proteins with levels of expression correlating with mCbr1 overexpression. The expression of voltage-gated anion channels, which may be directly related to calcium ion-related myocardial contraction, was also upregulated.
mCbr1 transgenic mice may be useful for further in vivo analyses of the molecular mechanisms regulated by Cbr1; such analyses will provide a better understanding of its effects on carcinogenesis and cardiotoxicity of certain cancer drugs.
羰基还原酶1（CBR1）是一种依赖烟酰胺腺嘌呤二核苷酸磷酸（NADPH）的还原酶，具有广泛的底物特异性。CBR1催化多种羰基化合物的还原，包括醌类、前列腺素、甲萘醌和多种外源性物质，同时还参与各种细胞过程，如致癌作用、细胞凋亡、信号转导和耐药性。在本研究中，我们旨在生成过表达小鼠Cbr1（mCbr1）的转基因小鼠，表征mCbr1在不同器官中的表达，并确定蛋白质表达模式的变化。
为了更深入地了解CBR1的功能，我们生成了全身过表达CBR1的转基因小鼠。这些转基因小鼠在CAG启动子下过表达3xFLAG标记的mCbr1（3xFLAG-mCbr1）。生成了两系转基因小鼠，一系在多个组织中表达3xFLAG-mCbr1，另一系在心脏中特异性表达3xFLAG-mCbr1。使用转基因小鼠心脏进行的通路和网络分析确定了73种蛋白质，其表达水平与mCbr1过表达相关。可能与钙离子相关的心肌收缩直接相关的电压门控阴离子通道的表达也上调。
mCbr1转基因小鼠可能有助于进一步体内分析由Cbr1调节的分子机制；此类分析将有助于更好地了解其对某些癌症药物致癌作用和心脏毒性的影响。
