title: In Vitro Methods for Studying the Mechanisms of Resistance to DNA-Damaging Therapeutic Drugs.
abstract: Most commonly used anticancer drugs exert their effects mainly by causing DNA damage. The enhancement in DNA damage response (DDR) is considered a key mechanism that enables cancer cells to survive through eliminating the damaged DNA lesions and thereby developing resistance to DNA-damaging agents. This chapter describes the four experimental approaches for studying DDR and genotoxic drug resistance, including the use of γ-H2AX and comet assays to monitor DNA damage and repair capacity as well as the use of clonogenic and β-galactosidase staining assays to assess long-term cell fate after DNA-damaging treatment. Finally, we also present examples of these methods currently used in our laboratory for studying the role of FOXM1 in DNA damage-induced senescence and epirubicin resistance.
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK.
authors.forename: Pasarat
authors.initials: P
authors.lastname: Khongkow
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK.
authors.forename: Anna W
authors.initials: AW
authors.lastname: Middleton
authors.forename: Anna K
authors.initials: AK
authors.lastname: Middleton
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK.
authors.forename: Jocelyn P-M
authors.initials: JP
authors.lastname: Wong
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK.
authors.forename: Navrohit K
authors.initials: NK
authors.lastname: Kandola
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK.
authors.forename: Mesayamas
authors.initials: M
authors.lastname: Kongsema
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK.
authors.forename: Gabriela Nestal
authors.initials: GN
authors.lastname: de Moraes
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK.
authors.forename: Ana R
authors.initials: AR
authors.lastname: Gomes
authors.affiliation: Department of Surgery and Cancer, Imperial College London, Hammersmith Hospital Campus, Du Cane Road, London, W12 0NN, UK. eric.lam@imperial.ac.uk.
authors.forename: Eric W-F
authors.initials: EW
authors.lastname: Lam
mesh_terms: D000970:Antineoplastic Agents / Q000494:pharmacology*; D002470:Cell Survival / Q000187:drug effects; D016922:Cellular Senescence / Q000187:drug effects; D020552:Comet Assay / Q000379:methods*; D004249:DNA Damage; D019008:Drug Resistance, Neoplasm; D004586:Electrophoresis; D005455:Fluorescent Antibody Technique / Q000379:methods*; D006657:Histones / Q000378:metabolism; D006801:Humans; D061986:MCF-7 Cells; D013194:Staining and Labeling; D001616:beta-Galactosidase / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; C468783:H2AX protein, human; D006657:Histones; D001616:beta-Galactosidase
keywords: Clonogenic assay; Comet assay; DNA damage; Resistance; β-Galactosidase staining; γ-H2AX