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A novel radiation-induced p53 mutation is not implicated in radiation resistance via a dominant-negative effect. PLoS One 2014;9(2):e87492

Date

02/22/2014

Pubmed ID

24558369

Pubmed Central ID

PMC3928108

DOI

10.1371/journal.pone.0087492

Abstract

Understanding the mutations that confer radiation resistance is crucial to developing mechanisms to subvert this resistance. Here we describe the creation of a radiation resistant cell line and characterization of a novel p53 mutation. Treatment with 20 Gy radiation was used to induce mutations in the H460 lung cancer cell line; radiation resistance was confirmed by clonogenic assay. Limited sequencing was performed on the resistant cells created and compared to the parent cell line, leading to the identification of a novel mutation (del) at the end of the DNA binding domain of p53. Levels of p53, phospho-p53, p21, total caspase 3 and cleaved caspase 3 in radiation resistant cells and the radiation susceptible (parent) line were compared, all of which were found to be similar. These patterns held true after analysis of p53 overexpression in H460 cells; however, H1299 cells transfected with mutant p53 did not express p21, whereas those given WT p53 produced a significant amount, as expected. A luciferase assay demonstrated the inability of mutant p53 to bind its consensus elements. An MTS assay using H460 and H1299 cells transfected with WT or mutant p53 showed that the novel mutation did not improve cell survival. In summary, functional characterization of a radiation-induced p53 mutation in the H460 lung cancer cell line does not implicate it in the development of radiation resistance.

Author List

Sun Y, Myers CJ, Dicker AP, Lu B

Author

Yunguang Sun MD, PhD Assistant Professor in the Pathology department at Medical College of Wisconsin




MESH terms used to index this publication - Major topics in bold

Cell Line, Tumor
Cell Survival
Gene Deletion
Gene Expression Regulation, Neoplastic
Genes, Dominant
Humans
Lung Neoplasms
Mutation
Protein Binding
Radiation Tolerance
Tumor Suppressor Protein p53
jenkins-FCD Prod-482 91ad8a360b6da540234915ea01ff80e38bfdb40a