Functional analyses reveal the essential role of SOX6 and RUNX2 in the communication of chondrocyte and osteoblast

This study provides novel evidence that sex determining region Y (SRY)-box (SOX6) and runt-related transcription factor 2 (RUNX2) play essential roles in the communication of chondrocyte and osteoblast. Our findings open a new avenue to the limited understanding of the coordination effect between chondrogenesis and osteogenesis. Sox6 and Runx2 are two new susceptibility genes for osteoporosis identified by genome-wide association studies, but the functions of these genes in osteogenesis remain unclear. Both genes are essential transcription factors in chondrogenesis, which reminds us that SOX6 and RUNX2 might be involved in the coordination of chondrogenesis and osteogenesis. Therefore, this study aimed to investigate the functions of SOX6 and RUNX2 in the coupling regulation of chondrogenesis and osteogenesis. We established a chondrogenic differentiation model of ATDC5 cell and profiled the expression of SOX6 and RUNX2 during chondroblast differentiation. We co-cultured osteoblast cells with ATDC5 cells in different differentiation stages and examined the proliferation, cell cycle progression, apoptosis, and differentiation of osteoblast cells. SOX6 or RUNX2 was knocked down using specific siRNA and the effect was examined. During chondrogenic differentiation, SOX6 and RUNX2 expressed sequentially in the proliferating and hypertrophic stages. Proliferative ATDC5 cells stimulated the multiplication of osteoblasts and promoted more osteoblasts to enter S-phase. Hypertrophic ATDC5 cells enhanced the differentiation of osteoblasts. Yet, the apoptosis of osteoblasts was neither affected by proliferating nor hypertrophic ATDC5 cells. Knockdown of SOX6 in proliferating ATDC5 cells significantly repressed the stimulation of osteoblast multiplication, whereas depletion of RUNX2 in hypertrophic ATDC5 cells retarded the expression of osteoblastic markers. Our study first reveals that SOX6 and RUNX2 play important roles in the chondrogenesis-osteogenesis coordination. This finding enriches the limited understanding about this coordination and unravels the novel function of SOX6 and RUNX2 in the endochondral ossification.