Effect of insulin on the mRNA expression of procollagen N-proteinases in chondrosarcoma OUMS-27 cells

Abstract

Chondrosarcoma is one of the most common bone tumors, and at present, there is no non-invasive treatment option for this cancer. The chondrosarcoma OUMS-27 cell line produces proteoglycan and type II, IX, and XI collagens, which constitutes cartilage tissue. A disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS) proteases are a group of secreted proteases, which include the procollagen N-proteinases ADAMTS-2, -3 and -14. These procollagen N-proteinases perform a role in the processing of procollagens to collagen and the maturation of type I collagen. The present study aimed to improve the understanding of the causes of metastasis, local invasion and resistance to chemo- and radiotherapy in chondrosarcoma, as well as the effect of insulin on cancer cells. The present study was designed to reveal the effects of insulin on procollagen N-proteinases in chondrosarcoma OUMS-27 cells. The cells were cultured in Dulbecco's modified Eagle's medium (DMEM) alone or in DMEM containing 10 µg/ml insulin. The medium was changed every other day for 11 days. The cells were harvested on days 1, 3, 7 and 11, and total RNA isolation was performed immediately following harvesting. The expression levels of ADAMTS2, ADAMTS3 and ADAMTS14 mRNA were estimated by reverse transcription-quantitative polymerase chain reaction using appropriate primers. ADAMTS2 mRNA expression was found to be decreased on day 7 (P=0.028) and increased at day 11 compared with the control group (P=0.016). The increase in mRNA concentration at day 11 was significantly different compared to the concentrations on days 3 (P=0.047) and 7 (P=0.008). The expression of ADAMTS3 mRNA decreased immediately subsequent to insulin induction on day 1 compared with the control group (P=0.008). The most evident decrease in mRNA concentration was seen at day 7 subsequent to insulin induction (P=0.008). The present results demonstrated that ADAMTS2 and ADAMTS3 may perform a role in the invasion and metastasis of tumors, and may also possess proteolytic activity that results in the breakdown of the extracellular matrix (ECM). Insulin itself can modulate the biosynthesis of ECM macromolecules that are altered in diabetes through various pathways.

IL-10 Could Play a Role in the Interrelation between Diabetes Mellitus and Osteoarthritis

“…Therefore, there is a high need for reliable and reproducible cell culture systems to avoid the problems of cell number limitations and high inter-donor variability. The chondrocyte cell line Okayama University Medical School (OUMS)-27 is derived from chondrosarcoma and expresses a differentiated chondrocytic phenotype offering the capacity to synthesise cartilage-specific ECM components such as proteoglycans (PG) and collagen (col) types II, IX and X [34,35,36]. Since OUMS-27 have been successfully used as a model to study the interrelation between chondrocytes and ECM, we examined this cell line for its suitability in OA research to investigate the interrelation of OA and T2DM.…”

Genetic sharing with coronary artery disease identifies potential novel loci for bone mineral density

“…Totally we identified 7 pleiotropic SNPs mapping to 3 chromosomes reached the significance threshold of ccFDR < 0.05 (Figure 2 and Table 1). Genes encompassed by these pleiotropic SNPs were found to be associated with bone metabolism [22,23] and/or CAD pathology [24], as well as risk factors for both osteoporosis and CAD, such as obesity and insulin resistance [25,26]. …”

“…SOST inhibits Wnt signaling and negatively regulates bone formation [34], TNFRSF11B is the decoy receptor for TNFSF11 and neutralizes its function in osteoclastogenesis [35]. Interestingly, though some genes encompassed by cFDR-significant SNPs did not have any interaction with other cFDR-significant genes in this network, they were found in studies to be associated with bone tumors (TOM1L2 and ADAMTS2) [22,23] and risk factors to osteoporosis, such as insulin resistance (TCERG1L) [36] and obesity (RAI1) [25]. …”

“…TOM1L2 was encompassed by rs12943500 and rs6502629, it was reported in a previous study that TOM1L2 was associated with 17p amplicons in osteosarcoma – a fatal malignant tumor of bone [23]. rs10039254 was located in the intron region of ADAMTS2, which was associated with the invasion and metastasis of chondrosarsoma – a common bone tumor [22], increased ADAMTS2 expression was detected in coronary plaques causing myocardial infarction [43]. rs10765090 was located near TCERG1L, in African Americans, TCERG1L was confirmed to be significantly associated with insulin resistance [26], which negatively affects bone mass and is an important risk factor for CAD.…”

Regeneration and healing of bone and cartilage in type-1 and type-2 diabetes: the effects of insulin

“…Many studies are focused on the effects of several molecules to elucidate the matrix biology of chondrocytes and pathophysiology of experimental osteoarthritis (Akyol et al, 2013(Akyol et al, , 2014b. A list of the beneficial effects of insulin (Akyol et al, 2015a;Altuntaş et al, 2015) discussed in detail via whether its regulating effect on hyperglycemia or its direct effect on cells may be as follows: accelerating fracture healing, improving resistance to fractures, providing growth and development of skeletal structures in children, and ensuring success with preoperational processes in dental implant surgery. Insulin does not necessarily participate in all stages of ADAMTS metabolism.…”