Antiproliferative MCR peptides block physical interaction of insulin with retinoblastoma protein (RB) in human lung cancer cells

Radulescu, Razvan Tudor
Kehe, Kai
Journal Title
Journal ISSN
Volume Title
Fifteen years ago, a structural analysis of the hormone insulin and the retinoblastoma tumor suppressor protein (RB) revealed that they may physically interact with one another. Subsequently, an RB peptide corresponding to the proposed RB binding site for insulin was found to recognize full-length insulin in vitro. As part of efforts aimed at developing this RB peptide into an anti-cancer drug, this molecule was chemically coupled to a cellular internalization signal and termed "MCR peptide". Meanwhile, several such MCR peptide variants have been demonstrated to restrain the proliferation of different human cancer cells in vitro and in vivo. Moreover, one of the MCR peptides coined MCR-10 was shown to be capable of interfering with the complex formation between insulin and RB in HepG2 human hepatoma cells, as monitored by immunofluorescence. This latter result indicating an in vivo association between insulin and RB was confirmed by a follow-up study combining the methods of co-immunoprecipitation and immunoblotting. Here, we provide evidence for the existence of the insulin-RB complex in A549 human non-small cell lung cancer cells. Specifically, we demonstrate this heterodimer by means of a magnetic beads-based immunoprecipitation approach and equally show that this dimer can be disrupted by MCR-4 or MCR-10 each of which is known to possess antiproliferative properties, yet to a much lesser extent by a control peptide. Thus, this investigation has yielded another important proof for the occurrence of the insulin-RB dimer and, furthermore, its validity as a target for antineoplastic MCR peptides.
Comment: 12 pages, 3 figures
Quantitative Biology - Subcellular Processes, Quantitative Biology - Biomolecules