Related Experiment Video
Updated: Apr 7, 2026
Isolation, Expansion, and Adipogenic Induction of CD34+CD31+ Endothelial Cells from Human Omental and Subcutaneous Adipose Tissue
Published on: July 17, 2018
The Investigation of ADAMTS16 in Insulin-Induced Human Chondrosarcoma Cells
Ozlem Cakmak1, Ismail Comertoglu2, Ridvan Firat3
11 Department of Biology Educations, Faculty of Education, Gazi University , Ankara, Turkey .
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.
Insights
Insulin treatment insignificantly decreased ADAMTS16 mRNA but gradually reduced ADAMTS16 protein in chondrosarcoma cells. This suggests insulin may regulate extracellular matrix turnover by affecting ADAMTS16 levels.
Area of Science:
- Biochemistry
- Cell Biology
- Extracellular Matrix Research
Background:
- Disintegrin-like metalloproteinases with thrombospondin motifs (ADAMTS) are secreted enzymes involved in extracellular matrix (ECM) remodeling.
- Insulin is known to stimulate proteoglycan biosynthesis in chondrocytes, key cells in cartilage formation and maintenance.
- Understanding the regulation of ADAMTS family members, like ADAMTS16, by insulin is crucial for comprehending ECM dynamics in conditions like chondrosarcoma.
Purpose of the Study:
- To investigate the time-dependent effects of insulin on ADAMTS16 expression in a human chondrosarcoma cell line (OUMS-27).
- To determine if insulin influences ADAMTS16 expression in parallel with its known effects on proteoglycan biosynthesis.
- To explore the potential role of insulin in regulating ECM turnover via ADAMTS16.
Main Methods:
- Human chondrosarcoma cells (OUMS-27) were cultured with or without insulin (10 μg/mL) for up to 11 days, with media changes every other day.
- Quantitative real-time PCR (qRT-PCR) was employed to measure ADAMTS16 mRNA expression levels at various time points.
- Western blotting was used to assess the protein levels of ADAMTS16 in response to insulin treatment over time.
Main Results:
- Insulin treatment resulted in an insignificant decrease in ADAMTS16 mRNA levels in chondrosarcoma cells.
- A progressive reduction in ADAMTS16 protein levels was observed in insulin-treated cells compared to control groups over the 11-day period.
- These findings indicate a differential impact of insulin on ADAMTS16 at the mRNA and protein levels.
Conclusions:
- Insulin may play a regulatory role in modulating ADAMTS16 expression and/or activity within chondrosarcoma cells.
- The observed changes in ADAMTS16 suggest a potential involvement of insulin in the dynamic processes of extracellular matrix turnover in these cells.
- Further research is warranted to elucidate the precise mechanisms by which insulin affects ADAMTS16 and its contribution to chondrosarcoma pathology.
