Nuclear colocalization and complex formation of insulin with retinoblastoma protein in HepG2 human hepatoma cells.
Previous structural and biochemical evidence had suggested that insulin may bind to the nuclear tumor suppressor retinoblastoma protein (RB). The present study is now the first to unravel the physical and functional interaction between a growth factor and an anti-oncoprotein, specifically demonstrating the association between insulin and RB in living cells and finding that this complex formation is relevant for cell division. Our immunofluorescence microscopy data suggest that insulin colocalizes with RB in the cell nuclei of HepG2 human hepatoma cells and that contacts the B-region of the RB pocket. Furthermore, these events were found to correlate with an enhancement of cell proliferation. These results are in line with the initial structure-based predictions and, moreover, provide a suitable starting point for the further understanding as well as the pharmacological modulation of nucleocrine interactions between growth factors and tumor suppressors, in physiology and disease.
Walther-Straub-Institut für Pharmakologie und Toxikologie, Ludwig-Maximilians-Universität, Nussbaumstrasse 26, 80336 Münchem, Germany.
R T
RT
Radulescu
E D
ED
Doklea
K
K
Kehe
H
H
Mückter
D000911:Antibodies, Monoclonal / Q000494:pharmacology; D001667:Binding, Competitive; D006528:Carcinoma, Hepatocellular / Q000378:metabolism*; D002455:Cell Division / Q000187:drug effects; D002467:Cell Nucleus / Q000378:metabolism*; D004396:Coloring Agents; D006801:Humans; D007328:Insulin / Q000378:metabolism* / Q000494:pharmacology; D008113:Liver Neoplasms / Q000378:metabolism*; D008856:Microscopy, Fluorescence; D016160:Retinoblastoma Protein / Q000276:immunology / Q000378:metabolism*; D013329:Structure-Activity Relationship; D013997:Time Factors; D014407:Tumor Cells, Cultured
D000911:Antibodies, Monoclonal; D004396:Coloring Agents; D007328:Insulin; D016160:Retinoblastoma Protein