Fluorescent peptide biosensor for monitoring CDK4/cyclin D kinase activity in melanoma cell extracts, mouse xenografts and skin biopsies

Publication date: 15 November 2016 Source:Biosensors and Bioelectronics, Volume 85 Author(s): Camille Prével, Morgan Pellerano, Juan A. González-Vera, Pauline Henri, Laurent Meunier, Julien Vollaire, Véronique Josserand, May C. Morris Melanoma constitutes the most aggressive form of skin cancer, which further metastasizes into a deadly form of cancer. The p16INK4a-Cyclin D-CDK4/6-pRb pathway is dysregulated in 90% of melanomas. CDK4/Cyclin D kinase hyperactivation, associated with mutation of CDK4, amplification of Cyclin D or loss of p16INK4a leads to increased risk of developing melanoma. This kinase therefore constitutes a key biomarker in melanoma and an emerging pharmacological target, however there are no tools enabling direct detection or quantification of its activity. Here we report on the design and application of a fluorescent peptide biosensor to quantify CDK4 activity in melanoma cell extracts, skin biopsies and melanoma xenografts. This biosensor provides sensitive means of comparing CDK4 activity between different melanoma cell lines and further responds to CDK4 downregulation by siRNA or small-molecule inhibitors. By affording means of monitoring CDK4 hyperactivity consequent to cancer-associated molecular alterations in upstream signaling pathways that converge upon this kinase, this biosensor offers an alternative to immunological identification of melanoma-specific biomarkers, thereby constituting an attractive tool for diagnostic pur...
Source: Biosensors and Bioelectronics - Category: Biotechnology Source Type: research