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Journal of Cancer Prevention

Original Article

Cancer prevention research 2007; 12(4): 310-318

Published online December 30, 2007

© Korean Society of Cancer Prevention

Cellular Proteins Related to Osteoclastogenesis in RAW 264.7 Cells

Meeyeon Yoo1, Yuna Youn1, Nari Lee2 and Soonyoung Choi1

Abstract

There are many factors related to osteoclastogenesis. Above all, chemokines and related ligands that are known to be related to immune response mechanisms have been studied by many scientists. This study investigates proteins that increase or decrease osteoclastogenesis. First, differentiation was induced in mouse monocyte/macrophage RAW 264.7 cells with TNF-related activation-induced cytokine (TRANCE, 400 ng/ml) and macrophage colony-stimulating factor (M-CSF, 50 ng/ml) for six days using purified total RNA. Differentially expressed gene screening was then done after synthesizing cDNA with total RNA. From the screening results, 10 differentially expressed genes were found during osteoclast differentiation. These were identified through a BLAST search. The V-ATPase and osteopontin that was identified by this screening is known to play an important role in bone resorption. Following the screening procedure, expression was confirmed by RT-PCR. In addition, other genes were identified via the BLAST search. Among them was RhoC, which is in the Ras homolog gene family (member C). The Ras family has been associated with the effects of tumor expression and differentiation, as well as with motility and resorption of osteoclasts. However, it is not known to be related to osteoclastogenesis. Finally, proteins related to RhoC were studied by immunoprecipitation during osteoclast differentiation. After a Maldi-TOF assay, it was determined that Ղ-actin is related to the actin-rich ring. Thus, further study of Ղ-actin is necessary. (Cancer Prev Res 12, 310-318, 2007)

Keywords: Osteoclast, Macrophage, Osteoclastogenesis, Differentially expressed gene screening

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