The discovery of agents which disrupt cancer cell division by specifically targeting key components of the cell-cycle machinery represents a major focus of recent drug discovery efforts in Oncology. The drug discovery process can be greatly enhanced by multiparametric cellular analysis which can assist in confirmation, often in a few multiplexed assays, of the mechanism of action (MOA) of compounds identified through biochemical screening or similar in vitro methods. High-Content Analysis (HCA) is a technique based on automated microscopy which enables multiparametric analysis of fluorescent indicators to define cellular responses to compound treatment. Several distinct fluorescence channels can be acquired and analyzed within a single measure in the same cell population. Here we present a multiparametric HCA approach to characterize potential cell-cycle inhibitors in osteosarcoma U-2 OS adherent cell cultures. This approach allows monitoring of compound-induced cell-cycle perturbations by analyzing specific cellular markers such as nuclear morphology, DNA content or histone H3 phosphorylation. Moreover, the induction of DNA damage response or apoptosis can also be readily evaluated. By considering the profile of the investigated cellular markers at different compound concentrations, a fingerprint defines the cellular and molecular phenotype associated with each compound.
Access this chapter
Tax calculation will be finalised at checkout
Purchases are for personal use only
Preview
Unable to display preview. Download preview PDF.
Author information
Authors and Affiliations
Editor information
Editors and Affiliations
Rights and permissions
Copyright information
© 2007 Springer
About this paper
Cite this paper
Gasparri, F., Ciavolella, A., Galvani, A. (2007). Cell-Cycle Inhibitor Profiling by High-Content Analysis. In: Fagagna, F.d.d., Chiocca, S., McBlane, F., Cavallaro, U. (eds) Advances in Molecular Oncology. Advances In Experimental Medicine And Biology, vol 604. Springer, Boston, MA. https://doi.org/10.1007/978-0-387-69116-9_13
Download citation
DOI: https://doi.org/10.1007/978-0-387-69116-9_13
Publisher Name: Springer, Boston, MA
Print ISBN: 978-0-387-69114-5
Online ISBN: 978-0-387-69116-9
eBook Packages: Biomedical and Life SciencesBiomedical and Life Sciences (R0)