Identification of insulin-regulated genes in the UMR-106 osteosarcoma cells.

A130

Recent studies have suggested that diabetic patients with insulin deficiency have bone metabolic disorders such as loss of bone density. These studies also indicate that insulin may have some role in bone cell metabolism. Thus, insulin-regulated gene expression was explored to determine if insulin exerts regulatory effects in osteosarcoma UMR-106 cells. UMR-106 cells were treated with insulin (100 nM) and total RNA was isolated. Differential Display PCR (DD-PCR) was carried out and followed by cloning, sequencing and searching the data base in order to identify the gene. By the DD-PCR experiments, it has been found that expression of 3 PCR products were increased by insulin whereas expression of 2 PCR products were decreased by insulin: the most significantly increased 1 PCR product in response to insulin was found and identified as "Rattus norvegicus (clone J13) ferritin light subunit mRNA, complete cds". The result from RT-PCR analysis shows that insulin significantly stimulated the mRNA expression of ferritin light chain. Insulin also stimilated up-regulated expression of ferritin light chain protein. These results suggest that insulin may play an important role in Fe metabolism in osteosarcoma cells.