The gene expression and protein profiles of ADAMTS and TIMP in human chondrosarcoma cell lines induced by insulin: The potential mechanisms for skeletal and articular abnormalities in diabetes.

Akyol, Sumeyya; Karagoz, Zehra; Inan, Nuran Dingil; Butun, Ilknur; Benli, Ismail; Demircan, Kadir; Yigitoglu, Muhammet Ramazan; Akyol, Omer; Sahin, Semsettin; Ozyurt, Huseyin

Background: The delay in wound healing, decrease in the long bones resilience to fracture, and delay in fracture healing are among common complications diabetes mellitus (DM) patients, and they still remain as challenging issues to be solved. The mechanism has not been fully understood yet, but high sugar and/or insulin deficiency or unresponsiveness to insulin in blood are potential causes to blame. Extracellular matrix degradation/remodeling is one of the important mechanisms whereby cell differentiation, bone remodeling and wound repair can be regulated. A disintegrin and metalloproteinase with a thrombospondin type 1 motif (ADAMTS) proteins play important roles in cartilage/bone metabolism. This study aimed to determine whether ADAMTS/Tissue inhibitors of metalloproteinases (TIMP) proteins were affected by insulin application in OUMS-27 (chondrosarcoma) cells. Material and Methods: OUMS-27 cells were induced by 10μg/mL insulin for 1, 3, 7, and 11 days. Cells were harvested, mRNA and protein extractions were performed. Total mRNA and cDNA levels were measured by qRT-PCR and protein levels were detected by WB. Results: ADAMTS1,5, and 7 levels were significantly decreased, while TIMP-3 levels were detected increased (mRNA/protein concentrations). Conclusion: Pathologies and disturbances of cartilage/bone metabolism, delayed fracture healing in particular, in patients with DM may result from insulin deficiency. ADAMTS genes that play a role in healing process are increased during insulin deficiency, which consequently interrupts healing process by causing cartilage extracellular matrix (ECM) degradation.

DIABETES complications; DNA analysis; EXPERIMENTAL design; IN vitro studies; KRUSKAL-Wallis Test; SKELETAL muscle; CELL culture; ARTICULAR ligaments; WESTERN immunoblotting; PROTEOLYTIC enzymes; MANN Whitney U Test; GENE expression; INSULIN; IMMUNOBLOTTING; CHONDROSARCOMA; GLYCOPROTEINS; TISSUE inhibitors of metalloproteinases; MESSENGER RNA; DESCRIPTIVE statistics; CELL lines; POLYMERASE chain reaction; DATA analysis software; FRACTURE healing

Electronic Journal of General Medicine, 2020, Vol 17, Issue 1, p1

2516-3507

Academic Journal

10.29333/ejgm/112767