Dual regulation of glycogen metabolism by insulin and insulin-like growth factors in human hepatoma cells (HEP-G2). Analysis with an anti-receptor monoclonal antibody.
Insulin and the insulinlike growth factors (IGF-I and IGF-II) are members of a family of hormones that regulate the metabolism and growth of many tissues. Cultured HEP-G2 cells (a minimal deviation human hepatoma) have insulin receptors and respond to insulin by increasing their glycogen metabolism. In the present study with HEP-G2 cells, we used 125I-labeled insulin, IGF-I, and IGF-II to identify distinct receptors for each hormone by competition-inhibition studies. Unlabeled insulin was able to inhibit 125I-IGF-I binding but not 125I-IGF-II binding. A mouse monoclonal antibody to the human insulin receptor that inhibits insulin binding and blocks insulin action inhibited 75% of 125I-insulin binding, but inhibited neither 125I-IGF-I nor 125I-IGF-II binding. When glycogen metabolism was studied, insulin stimulated [3H]glucose incorporation into glycogen in a biphasic manner; one phase that was 20-30% of the maximal response occurred over 1-100 pM, and the other phase occurred over 100 pM-100 nM. The anti-receptor monoclonal antibody inhibited the first phase of insulin stimulation but not the second. Both IGF-I and IGF-II stimulated [3H]glucose incorporation over the range of 10 pM-10 nM; IGF-I was three to fivefold more potent. The monoclonal antibody, however, was without effect on IGF regulation of glycogen metabolism. Therefore, these studies indicate that insulin as well as the IGFs at physiological concentrations regulate glycogen metabolism in HEP-G2 cells. Moreover, this regulation of glycogen metabolism is mediated by both the insulin receptor and the IGF receptors.
D000818:Animals; D000911:Antibodies, Monoclonal / Q000502:physiology; D001667:Binding, Competitive; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D002460:Cell Line; D005947:Glucose / Q000378:metabolism; D006003:Glycogen / Q000378:metabolism*; D006801:Humans; D007328:Insulin / Q000378:metabolism / Q000502:physiology*; D008113:Liver Neoplasms / Q000378:metabolism*; D051379:Mice; D010455:Peptides / Q000378:metabolism / Q000502:physiology*; D011972:Receptor, Insulin / Q000032:analysis / Q000276:immunology; D011956:Receptors, Cell Surface / Q000032:analysis; D017451:Receptors, Somatomedin; D013002:Somatomedins / Q000378:metabolism / Q000502:physiology*
D000911:Antibodies, Monoclonal; D007328:Insulin; D010455:Peptides; D011956:Receptors, Cell Surface; D017451:Receptors, Somatomedin; D013002:Somatomedins; D006003:Glycogen; D011972:Receptor, Insulin; D005947:Glucose