Transglutaminase-2 Is Involved in Expression of Osteoprotegerin in MG-63 Osteosarcoma Cells.
- Author:
Hye Ja LEE
1
;
Chang Hoon LEE
Author Information
1. College of Pharmacy, Dongguk University, Seoul 100-715, Republic of Korea. uatheone@dongguk.edu
- Publication Type:Original Article
- Keywords:
Osteoprotegerin;
Transglutaminase-2;
MG-63 cell;
Cystamine;
IL-1beta;
TPA
- MeSH:
Blotting, Western;
Bone Remodeling;
Cystamine;
Gene Silencing;
Glycoproteins;
Interleukin-1beta;
Multifunctional Enzymes;
Neoplasm Metastasis;
Osteogenesis;
Osteoprotegerin*;
Osteosarcoma*;
Receptors, Tumor Necrosis Factor
- From:Biomolecules & Therapeutics
2013;21(3):204-209
- CountryRepublic of Korea
- Language:English
-
Abstract:
Osteoprotegerin (OPG) is a secreted glycoprotein and a member of the tumor necrosis factor receptor superfamily. It usually functions in bone remodeling, by inhibiting osteoclastogenesis through interaction with a receptor activator of the nuclear factor kappaB (RANKL). Transglutaminases-2 (Tgase-2) is a group of multifunctional enzymes that plays a role in cancer cell metastasis and bone formation. However, relationship between OPG and Tgase-2 is not studied. Therefore, we investigated the involvement of 12-O-Tetradecanoylphorbol 13-acetate in the expression of OPG in MG-63 osteosarcoma cells. Interleukin-1beta time-dependently induced OPG and Tgase-2 expression in cell lysates and media of the MG-63 cells by a Western blot. Additional 110 kda band was found in the media of MG-63 cells. 12-O-Tetradecanoylphorbol 13-acetate also induced OPG and Tgase-2 expression. However, an 110 kda band was not found in TPA-treated media of MG-63 cells. Cystamine, a Tgase-2 inhibitor, dose-dependently suppressed the expression of OPG in MG-63 cells. Gene silencing of Tgase-2 also significantly suppressed the expression of OPG in MG-63 cells. Next, we examined whether a band of 110 kda of OPG contains an isopeptide bond, an indication of Tgase-2 action, by monoclonal antibody specific for the isopeptide bond. However, we could not find the isopeptide bond at 110 kda but 77 kda, which is believed to be the band position of Tgase-2. This suggested that 110 kda is not the direct product of Tgase-2's action. All together, OPG and Tgase-2 is induced by IL-1beta or TPA in MG-63 cells and Tgase-2 is involved in OPG expression in MG-63 cells.
