Ozlem Cakmak1, Ismail Comertoglu2, Ridvan Firat3, Haci Kemal Erdemli4, S Fatih Kursunlu5, Sumeyya Akyol6, Veli Ugurcu7, Aynur Altuntas8, Bahattin Adam9, Kadir Demircan6. 1. 1 Department of Biology Educations, Faculty of Education, Gazi University , Ankara, Turkey . 2. 2 Department of Medical Genetics, Faculty of Medicine, Mevlana University , Konya, Turkey . 3. 3 Division of Medical Biochemistry Laboratory, Golbasi State Hospital , Ankara, Turkey . 4. 4 Department of Biochemistry Laboratory, Corum Training and Research Hospital , Corum, Turkey . 5. 5 Department of Periodontology, Faculty of Dentistry, Adnan Menderes University , Aydın, Turkey . 6. 6 Department of Medical Biology, Faculty of Medicine, Turgut Ozal University , Ankara, Turkey . 7. 7 Department of Medical Biochemistry, Dumlupinar University Medical Faculty , Kutahya, Turkey . 8. 8 Division of Chemistry, Ankara Regional Office of Council of Forensic Medicine , Ankara, Turkey . 9. 9 University of California Davis Medical School , Department of Biochemistry and Molecular Medicine, Sacramento, California.
Abstract
OBJECTIVES: A disintegrin-like metalloproteinase with thrombospondin motifs (ADAMTS) is a group of proteins that have enzymatic activity secreted by cells to the outside extracellular matrix. Insulin induces proteoglycan biosynthesis in chondrosarcoma chondrocytes. The purpose of the present in vitro study is to assess the time course effects of insulin on ADAMTS16 expression in OUMS-27 (human chondrosarcoma) cell line to examine whether insulin regulates ADAMTS16 expression as well as proteoglycan biosynthesis with multifaceted properties or not. METHODS: Chondrosarcoma cells were cultured in Dulbecco's modified Eagle's medium having either 10 μg/mL insulin or not. While the experiment was going on, the medium containing insulin had been changed every other day. Cells were harvested at 1st, 3rd, 7th, and 11th days; subsequently, RNA and proteins were isolated in every experimental group according to their time interval. RNA expression of ADAMTS was estimated by quantitative real-time polymerase chain reaction (qRT-PCR) by using primers. Immunoreactive protein levels were encountered by the western blot protein detection technique by using proper anti-ADAMTS16 antibodies. RESULTS: ADAMTS16 mRNA expression level of chondrosarcoma cells was found to be insignificantly decreased in chondrosarcoma cells induced by insulin detected by the qRT-PCR instrument. On the other hand, there was a gradual decrease in immune-reactant ADAMTS16 protein amount by the time course in insulin-treated cell groups when compared with control cells. CONCLUSION: It has been suggested that insulin might possibly regulate ADAMTS16 levels/activities in OUMS-27 chondrosarcoma cells taking a role in extracellular matrix turnover.
OBJECTIVES: A disintegrin-like metalloproteinase with thrombospondin motifs (ADAMTS) is a group of proteins that have enzymatic activity secreted by cells to the outside extracellular matrix. Insulin induces proteoglycan biosynthesis in chondrosarcoma chondrocytes. The purpose of the present in vitro study is to assess the time course effects of insulin on ADAMTS16 expression in OUMS-27 (humanchondrosarcoma) cell line to examine whether insulin regulates ADAMTS16 expression as well as proteoglycan biosynthesis with multifaceted properties or not. METHODS:Chondrosarcoma cells were cultured in Dulbecco's modified Eagle's medium having either 10 μg/mL insulin or not. While the experiment was going on, the medium containing insulin had been changed every other day. Cells were harvested at 1st, 3rd, 7th, and 11th days; subsequently, RNA and proteins were isolated in every experimental group according to their time interval. RNA expression of ADAMTS was estimated by quantitative real-time polymerase chain reaction (qRT-PCR) by using primers. Immunoreactive protein levels were encountered by the western blot protein detection technique by using proper anti-ADAMTS16 antibodies. RESULTS:ADAMTS16 mRNA expression level of chondrosarcoma cells was found to be insignificantly decreased in chondrosarcoma cells induced by insulin detected by the qRT-PCR instrument. On the other hand, there was a gradual decrease in immune-reactant ADAMTS16 protein amount by the time course in insulin-treated cell groups when compared with control cells. CONCLUSION: It has been suggested that insulin might possibly regulate ADAMTS16 levels/activities in OUMS-27 chondrosarcoma cells taking a role in extracellular matrix turnover.
Entities:
Keywords:
ADAMTS16; OUMS-27; RNA; chondrosarcoma; insulin; protein
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