MicroRNA-124 inhibits canine mammary carcinoma cell proliferation, migration and invasion by targeting CDH2
Xiaoli Ren, Yuying Fan, Dongmei Shi, Enshuang Xu, Yun Liu
Abstract
Canine mammary carcinoma (CMC) is the most common malignant tumor and the second leading cause of cancer-related mortality of dogs worldwide. MicroRNA-124 (miR-124) is an important tumor suppressor implicated in various aspects of carcinogenesis. However, the roles and mechanisms of miR-124 in CMC development remains to be determined. We used the quantitative real-time polymerase chain reaction (qRT-PCR) assay to evaluate the expression levels of miR-124 in CMC tissues obtained from 20 CMC cases and CHMm and CHMp cells. CMC cell lines were transfected with lipfactormine™2000, and the cell proliferation was measured by Cell Counting Kit-8 (CCK-8). Transwell assay were employed for evaluating the cell invasion and migration, while western blot assay was used to detect the protein changes in epithelial-mesenchymal transition (EMT) and CDH2 protein levels. The relationship between miR-124 and the 3'-untranslated region (3'-UTR) of CDH2 was predicted via bioinformatics analysis and verified by dual-luciferase reporter assay. The results revealed that miR-124 was reduced in CMC tissues and cell lines. Besides, observed high histological grade and tumor metastasis were associated with the down-regulation of miR-124 and up-regulation of CDH2. Functional analyses showed that in vitro transfection of CHMm and CHMp cells with miR-124 mimics inhibited their proliferation, migration, invasion, and EMT; however, transfection with miR-124 inhibitor resulted in the reversed effect. Besides, we showed that miR-124 directly suppressed the expression of CDH2, leading to the inhibition of CHMm cell proliferation and EMT. In conclusion, miR-124 regulates CMC tumor growth and EMT by targeting CDH2, maybe a potential therapeutic strategy against CMC.