title: Adaptation of cancer cells from different entities to the MDM2 inhibitor nutlin-3 results in the emergence of p53-mutated multi-drug-resistant cancer cells.
abstract: Six p53 wild-type cancer cell lines from infrequently p53-mutated entities (neuroblastoma, rhabdomyosarcoma, and melanoma) were continuously exposed to increasing concentrations of the murine double minute 2 inhibitor nutlin-3, resulting in the emergence of nutlin-3-resistant, p53-mutated sublines displaying a multi-drug resistance phenotype. Only 2 out of 28 sublines adapted to various cytotoxic drugs harboured p53 mutations. Nutlin-3-adapted UKF-NB-3 cells (UKF-NB-3(r)Nutlin(10 μM), harbouring a G245C mutation) were also radiation resistant. Analysis of UKF-NB-3 and UKF-NB-3(r)Nutlin(10 μM) cells by RNA interference experiments and lentiviral transduction of wild-type p53 into p53-mutated UKF-NB-3(r)Nutlin(10 μM) cells revealed that the loss of p53 function contributes to the multi-drug resistance of UKF-NB-3(r)Nutlin(10 μM) cells. Bioinformatics PANTHER pathway analysis based on microarray measurements of mRNA abundance indicated a substantial overlap in the signalling pathways differentially regulated between UKF-NB-3(r)Nutlin(10 μM) and UKF-NB-3 and between UKF-NB-3 and its cisplatin-, doxorubicin-, or vincristine-resistant sublines. Repeated nutlin-3 adaptation of neuroblastoma cells resulted in sublines harbouring various p53 mutations with high frequency. A p53 wild-type single cell-derived UKF-NB-3 clone was adapted to nutlin-3 in independent experiments. Eight out of ten resulting sublines were p53-mutated harbouring six different p53 mutations. This indicates that nutlin-3 induces de novo p53 mutations not initially present in the original cell population. Therefore, nutlin-3-treated cancer patients should be carefully monitored for the emergence of p53-mutated, multi-drug-resistant cells.
authors.affiliation: Institut für Medizinische Virologie, Klinikum der Goethe-Universität, Frankfurt am Main, Germany.
authors.forename: M
authors.initials: M
authors.lastname: Michaelis
authors.forename: F
authors.initials: F
authors.lastname: Rothweiler
authors.forename: S
authors.initials: S
authors.lastname: Barth
authors.forename: J
authors.initials: J
authors.lastname: Cinatl
authors.forename: M
authors.initials: M
authors.lastname: van Rikxoort
authors.forename: N
authors.initials: N
authors.lastname: Löschmann
authors.forename: Y
authors.initials: Y
authors.lastname: Voges
authors.forename: R
authors.initials: R
authors.lastname: Breitling
authors.forename: A
authors.initials: A
authors.lastname: von Deimling
authors.forename: F
authors.initials: F
authors.lastname: Rödel
authors.forename: K
authors.initials: K
authors.lastname: Weber
authors.forename: B
authors.initials: B
authors.lastname: Fehse
authors.forename: E
authors.initials: E
authors.lastname: Mack
authors.forename: T
authors.initials: T
authors.lastname: Stiewe
authors.forename: H W
authors.initials: HW
authors.lastname: Doerr
authors.forename: D
authors.initials: D
authors.lastname: Speidel
authors.forename: J
authors.initials: J
authors.lastname: Cinatl
mesh_terms: D000220:Adaptation, Biological / Q000187:drug effects; D000970:Antineoplastic Agents / Q000494:pharmacology*; D053148:Caspase 3 / Q000378:metabolism; D053179:Caspase 7 / Q000378:metabolism; D019008:Drug Resistance, Neoplasm / Q000235:genetics*; D006801:Humans; D007093:Imidazoles / Q000494:pharmacology*; D009154:Mutation; D010879:Piperazines / Q000494:pharmacology*; D051736:Proto-Oncogene Proteins c-mdm2 / Q000037:antagonists & inhibitors* / Q000378:metabolism; D034622:RNA Interference; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
chemical_list: D000970:Antineoplastic Agents; D007093:Imidazoles; D010879:Piperazines; D016159:Tumor Suppressor Protein p53; C482205:nutlin 3; C497206:MDM2 protein, human; D051736:Proto-Oncogene Proteins c-mdm2; D053148:Caspase 3; D053179:Caspase 7