title: Quantitative assessment of the complex dynamics of G1, S, and G2-M checkpoint activities.
abstract: Although studies of cell cycle perturbation and growth inhibition are common practice, they are unable to properly measure the activity of cell cycle checkpoints and frequently convey misinterpretation or incomplete pictures of the response to anticancer treatment. A measure of the strength of the treatment response of all checkpoints, with their time and dose dependence, provides a new way to evaluate the antiproliferative activity of the drugs, fully accounting for variation of the cell fates within a cancer cell line. This is achieved with an interdisciplinary approach, joining information from independent experimental platforms and interpreting all data univocally with a simple mathematical model of cell cycle proliferation. The model connects the dynamics of checkpoint activities at the molecular level with population-based flow cytometric and growth inhibition time course measures. With this method, the response to five drugs, characterized by different molecular mechanisms of action, was studied in a synoptic way, producing a publicly available database of time course measures with different techniques in a range of drug concentrations, from sublethal to frankly cytotoxic. Using the computer simulation program, we were able to closely reproduce all the measures in the experimental database by building for each drug a scenario of the time and dose dependence of G(1), S, and G(2)-M checkpoint activities. We showed that the response to each drug could be described as a combination of a few types of activities, each with its own strength and concentration threshold. The results gained from this method provide a means for exploring new concepts regarding the drug-cell cycle interaction.
authors.affiliation: Biophysics Unit, Laboratory of Anticancer Pharmacology, Department of Oncology, Istituto di Ricerche Farmacologiche Mario Negri, Milan, Italy. ubezio@marionegri.it
authors.forename: Paolo
authors.initials: P
authors.lastname: Ubezio
authors.forename: Monica
authors.initials: M
authors.lastname: Lupi
authors.forename: Davide
authors.initials: D
authors.lastname: Branduardi
authors.forename: Paolo
authors.initials: P
authors.lastname: Cappella
authors.forename: Edoardo
authors.initials: E
authors.lastname: Cavallini
authors.forename: Valentina
authors.initials: V
authors.lastname: Colombo
authors.forename: Giada
authors.initials: G
authors.lastname: Matera
authors.forename: Claudia
authors.initials: C
authors.lastname: Natoli
authors.forename: Daniela
authors.initials: D
authors.lastname: Tomasoni
authors.forename: Maurizio
authors.initials: M
authors.lastname: D'Incalci
mesh_terms: D002455:Cell Division; D002460:Cell Line; D003198:Computer Simulation; D005434:Flow Cytometry; D016193:G1 Phase; D016195:G2 Phase; D006801:Humans; D009369:Neoplasms / Q000473:pathology; D016196:S Phase