Sei-Myoung Han1, Chong-Woo Park1, Jin-Ok Ahn1, Sang-Chul Park1, Woo-Sung Jung1, Kyoung-Won Seo2, Jeong-Chan Ra3, Sung-Keun Kang3, Hee-Woo Lee4, Hwa-Young Youn4. 1. Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea. 2. Department of Veterinary Internal Medicine, College of Veterinary Medicine, Chungnam National University, Daejeon, Republic of Korea. 3. Biostar Stem Cell Research Center, K-STEMCELL Co. Ltd., Seoul, Republic of Korea. 4. Department of Veterinary Internal Medicine, College of Veterinary Medicine, Seoul National University, Seoul, Republic of Korea hyyoun@snu.ac.kr.
Abstract
BACKGROUND: Canine melanoma is the most common type of tumor in dogs. We investigated the effects of canine interferon-beta (cIFN-β)-overexpressing adipose tissue-derived mesenchymal stem cells (cATMSCs) on apoptosis and proliferation of canine melanoma cells. MATERIALS AND METHODS: Expression of IFN-β in cATMSCs was confirmed using reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assays. Flow cytometry was performed for cell-cycle analysis and apoptotic cell quantification of LMeC (melanoma) cells. Protein expression of cyclin D1, procaspase-3, activated caspase-3, and Bcl-2 homologous antagonist killer (Bak) was evaluated by western blot analysis. RESULTS: Decreased proportions of cells in S- and G0/G1 phases were observed in parallel with decreased cyclin D1 expression in LMeC cells treated with cIFN-β-cATMSC-conditioned media. Protein expression of active forms of caspase 3 and Bak increased in response to treatment with cIFN-β-cATMSC-conditioned media. CONCLUSION: IFN-β overexpression by cATMSCs was associated with pro-apoptotic and growth-inhibitory effects on canine melanoma cells. The antitumor effects of these cells have therapeutic potential for the treatment of canine melanoma. Copyright
BACKGROUND:Caninemelanoma is the most common type of tumor in dogs. We investigated the effects of canine interferon-beta (cIFN-β)-overexpressing adipose tissue-derived mesenchymal stem cells (cATMSCs) on apoptosis and proliferation of caninemelanoma cells. MATERIALS AND METHODS: Expression of IFN-β in cATMSCs was confirmed using reverse transcription-polymerase chain reaction and enzyme linked immunosorbent assays. Flow cytometry was performed for cell-cycle analysis and apoptotic cell quantification of LMeC (melanoma) cells. Protein expression of cyclin D1, procaspase-3, activated caspase-3, and Bcl-2 homologous antagonist killer (Bak) was evaluated by western blot analysis. RESULTS: Decreased proportions of cells in S- and G0/G1 phases were observed in parallel with decreased cyclin D1 expression in LMeC cells treated with cIFN-β-cATMSC-conditioned media. Protein expression of active forms of caspase 3 and Bak increased in response to treatment with cIFN-β-cATMSC-conditioned media. CONCLUSION: IFN-β overexpression by cATMSCs was associated with pro-apoptotic and growth-inhibitory effects on caninemelanoma cells. The antitumor effects of these cells have therapeutic potential for the treatment of caninemelanoma. Copyright