Insulin receptors and bioresponses in a human liver cell line (Hep G-2).
A newly developed human hepatoma cell line, designated Hep G-2, expresses high-affinity insulin receptors meeting all the expected criteria for classic insulin receptors. 125I-insulin binding is time-dependent and temperature-dependent and unlabeled insulin competes for the labeled hormone with a half-maximal displacement of 1-3 ng/ml. This indicates a Kd of about 10(-10) M. Since Scatchard analysis of the binding data results in a curvilinear plot and unlabeled insulin accelerates the dissociation of bound hormone, these receptors exhibit the negative cooperative interactions characteristic of insulin receptors in many other cell and tissue types. Proinsulin and des(Ala, Asp)-insulin compete for 125I-insulin binding with 4% and 2%, respectively, of the potency of insulin. Anti-(insulin receptor) antibody competes fully for insulin binding. The two insulin-like growth factors, multiplication-stimulating activity and IGF-I are 2% as potent as insulin against the Hep G-2 insulin receptor. Furthermore, Hep G-2 cells respond to insulin in several bioassays. Glucose uptake, glycogen synthase, uridine incorporation into RNA and acetate incorporation into lipid are all stimulated to varying degrees by physiological concentrations of insulin. In addition, these cells 'down-regulate' their insulin receptor, internalize 125I-insulin and degrade insulin in a manner similar to freshly isolated rodent hepatocytes. This is the first available human liver cell line in permanent culture in which both insulin receptors and biological responses have been carefully examined.
J M
JM
Podskalny
S
S
Takeda
R E
RE
Silverman
D
D
Tran
J L
JL
Carpentier
L
L
Orci
P
P
Gorden
D002460:Cell Line; D005947:Glucose / Q000378:metabolism; D006801:Humans; D007328:Insulin / Q000378:metabolism / Q000494:pharmacology*; D007457:Iodine Radioisotopes; D008055:Lipids / Q000096:biosynthesis; D008113:Liver Neoplasms / Q000032:analysis* / Q000378:metabolism / Q000473:pathology; D012313:RNA / Q000096:biosynthesis; D011972:Receptor, Insulin / Q000032:analysis*
D007328:Insulin; D007457:Iodine Radioisotopes; D008055:Lipids; D012313:RNA; D011972:Receptor, Insulin; D005947:Glucose