Methods: MC3T3-E1 osteoprogenitor cells were subjected to osteogenic medium, and the relationship between the endosome chloride channels ClC-3, ClC-4, ClC-5 and osteogenesis markers (alkaline phosphatase (ALP), osteocalcin (OC), bone sialoprotein (BSP), and runt-related transcription factor 2 (Runx2)) were analyzed with real-time PCR method. And MC3T3-E1 cells were transient transfected with Clcn3, Clcn4 or Clcn5 gene. The mRNA levels of ALP, OC, BSP, and Runx2 were compared between transfected cells and the control with real-time PCR method.
Results: The mRNA levels of ALP, OC, BSP and Runx2 increased after osteogenic induction, while those of Clcn3,Clcn4, Clcn5 were also increased. The overexpression of Clcn3, Clcn4 or Clcn5 gene in MC3T3-E1 cells upregulated the mRNA levels of ALP, OC, BSP, and Runx2 significantly.
Conclusion: we concluded that ClC-3, -4 and -5 chloride channels may play important roles in osteogenesis in MC3T3-E1 osteoprogenitor cells.