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Runx2 is required for chondrocyte proliferation and maturation. In the search of Runx2 target genes in chondrocytes, we found that Runx2 up-regulated the expression of hematopoietic cell kinase ( Hck) , which is a member of the Src tyrosine kinase family, in chondrocytes, that Hck expression was high in cartilaginous limb skeletons of wild-type mice but low in those of Runx2  -/- mice, and that Runx2 bound the promoter region of Hck . To investigate the functions of Hck in chondrocytes, transgenic mice expressing a constitutively active form of Hck ( Hck  CA ) were generated using the Col2a1 promoter/enhancer. The hind limb skeletons were fused, the tibia became a large, round mass, and the growth plate was markedly disorganized. Chondrocyte maturation was delayed until E16.5 but accelerated thereafter. BrdU-labeled, but not terminal deoxynucleotidyl transferase-mediated dUTP nick end labeling (TUNEL)-positive, chondrocytes were increased. Furthermore, Hck knock-down reduced the proliferation of primary chondrocytes. In microarray and real-time RT-PCR analyses using hind limb RNA from Hck  CA transgenic mice, the expression of Wnt ( Wnt10b , Tcf7 , Lef1 , Dkk1 ) and hedgehog ( Ihh , Ptch1 , and Gli1 ) signaling pathway genes was upregulated. These findings indicated that Hck , whose expression is regulated by Runx2, is highly expressed in chondrocytes, and that Hck  CA activates Wnt and hedgehog signaling pathways, and promotes chondrocyte proliferation without increasing apoptosis.

international journal of molecular sciences

Expression of a Constitutively Active Form of Hck in Chondrocytes Activates Wnt and Hedgehog Signaling Pathways, and Induces Chondrocyte Proliferation in Mice