Genetic studies revealed a crucial role of Distal-homebox (Dlx) genes in skeletal development, and our previous study demonstrated overexpressing DIx2 in neural crest cells led to abnormal cartilage structure, including ectopic cartilage in the maxillary region and nasal bone in mice. The aim of this study was to investigate how Dlx2 overexpression affects chondrogenesis in mouse chondroblast cell line TMC23 and the underlying mechanism. We first demonstrated that Dlx2 expression was upregulated during chondrogenesis in TMC23 cells. Moreover, forced overexpression of Dlx2 in TMC23 cells led to increased accumulation of aggrecan and type II collagen, markers of early chondrocyte differentiation, but had little effect on mRNA and protein levels of Aggrecan and Col2 alpha l, type II collagen gene. Importantly, Dlx2 overexpression decreased mRNA and protein levels of MMP13, a major collagenase degrading aggrecan and type II collagen during late stages of chondrogenesis. Luciferase-reporter and Chromatin-immunoprecipitation analysis demonstrated that MMP13 promoter contained two Dlx2-response elements, and Dlx2 inhibited MMP13 expression by directly binding to these two elements. Based on these observations, we propose that forced overexpression of Dlx2 enhances early chondrocyte differentiation by increasing accumulation of type II collagen and aggrecan, but interferes later stages of chondrocyte differentiation through inhibiting MMP13 expression. (C) 2018 Elsevier Inc. All rights reserved.