Zhang Bohan, He Lixin, Wang Xiao, Liu Wenjing, Li Yuxin, Wang Yinan, Feng Huili, Li Wenxuan, Qiu Changwei
Department of Clinical Veterinary Medicine, College of Veterinary Medicine, Huazhong Agricultural University, Wuhan 430070, China.
Cells. 2025 Jun 25;14(13):974. doi: 10.3390/cells14130974.
Canine mammary tumors (CMTs) are the common tumors in female dogs, and approximately 50% of CMTs are malignant tumors, with abnormal regulation of non-coding RNAs being a critical factor in disease progression. Currently, research on long non-coding RNAs (lncRNAs) regulating CMT development remains limited. This study identified a novel lncRNA, aiming to explore the role of lncRNA LOC610012 in CMTs. In this study, immunofluorescence and Western blot analyses were employed to detect protein expression. LncRNA LOC610012 is downregulated in CMT tissues and cells. Stable cells of LOC610012 were constructed by the lentivirus technique. Through a variety of experimental methods, LOC610012 inhibited the proliferation, invasion, and metastasis of CMT cells in in vitro and in vivo experiments conducted using cell culture and mouse models. Mechanistically, LOC610012 regulated the expression of EP3 and GSK-3β by targeting PTGS2, resulting in excessive production of reactive oxygen species (ROS), which inhibited cell viability. Similarly, through transmission electron microscopy, mitochondrial damage caused by LOC610012 was observed in CMT cells, which was manifested as mitochondrial swelling, membrane rupture, and mitochondrial ridge disappearance. PTGS2 could partially restore the inhibition of LOC610012 on cell activity. LOC610012 acts as a tumor suppressor gene in CMTs and as a potential biomarker for the disease.
犬乳腺肿瘤(CMTs)是雌性犬常见的肿瘤,约50%的CMTs为恶性肿瘤,非编码RNA的异常调控是疾病进展的关键因素。目前,关于长链非编码RNA(lncRNAs)调控CMT发生发展的研究仍然有限。本研究鉴定了一种新型lncRNA,旨在探讨lncRNA LOC610012在CMTs中的作用。在本研究中,采用免疫荧光和蛋白质印迹分析来检测蛋白质表达。lncRNA LOC610012在CMT组织和细胞中表达下调。通过慢病毒技术构建了LOC610012稳定细胞株。通过多种实验方法,在使用细胞培养和小鼠模型进行的体外和体内实验中,LOC610012抑制了CMT细胞的增殖、侵袭和转移。机制上,LOC610012通过靶向PTGS2调节EP3和GSK-3β的表达,导致活性氧(ROS)过量产生,从而抑制细胞活力。同样,通过透射电子显微镜观察到CMT细胞中存在由LOC610012引起的线粒体损伤,表现为线粒体肿胀、膜破裂和线粒体嵴消失。PTGS2可以部分恢复LOC610012对细胞活性的抑制作用。LOC610012在CMTs中作为肿瘤抑制基因发挥作用,并且是该疾病的潜在生物标志物。