Lymphoblastoid cell lines differing in p53 status show clear differences in basal gene expression with minor changes after irradiation

被引:18
作者
Zschenker, Oliver
Borgmann, Kerstin
Streichert, Thomas
Meier, Ingo
Wrona, Agnieszka
Dikomey, Ekkehard
机构
[1] Univ Hamburg, Hosp Eppendorf, Clin Radiotherapy & Radiooncol, Lab Radiobiol & Expt Radiooncol, D-20246 Hamburg, Germany
[2] Univ Hamburg, Hosp Eppendorf, Inst Clin Chem, Microarray Facil, D-20246 Hamburg, Germany
[3] Univ Hamburg, Hosp Eppendorf, Ctr Mol Neurobiol, D-20246 Hamburg, Germany
关键词
gene expression profile; industrial radiosensitivity; prediction; gene array; affymetrix; DOUBLE-STRAND BREAKS; IONIZING-RADIATION; CLONOGENIC SURVIVAL; MICROARRAY ANALYSIS; HUMAN FIBROBLASTS; TUMOR-SUPPRESSOR; GROWTH-FACTOR; DNA-REPAIR; WILD-TYPE; APOPTOSIS;
D O I
10.1016/j.radonc.2006.07.019
中图分类号
R73 [肿瘤学];
学科分类号
100214 ;
摘要
Background and purpose: The genetic profile as determined by microarray is considered to be an ideal marker of the individual radiosensitivity. However, it is still an open question, whether this profile has to be determined prior to or only after irradiation, since the expression of some genes is affected by irradiation. These changes are induced mainly due to a p53-dependent transactivation. Materials and methods: In this study gene expression profiles were measured for 3 lymphoblastoid cell lines differing in p53 status (p53 wt: TK6; p53null: TK6E6, p53mut: WTK1) measured either prior to or 3 h after exposure to 2 Gy. The gene expression profile was determined using the Affymetrix Human HG U133A GeneChip and for selective genes, variation in gene expression was validated by qRT-PCR. In addition, different assays were used to characterize the radioresponse of these three strains. Results: The three strains were found to be different in all aspects of radiosensitivity studied. Cells with p53wt showed more apoptosis, slightly stronger arrest in G1, but less lethal aberrations and a lower viability when compared to cells with mutated p53, whereas cells absent in p53 are characterized by an intermediate response. The gene expression profile measured prior to irradiation already revealed huge differences. Significance analysis of microarrays (SAM) identified 141 genes that changed expression twofold or more with a false discovery rate (FDR) of 5.4%. When compared to p53null cell line with p53wt showed a twofold difference in up- or down-regulation in 28 genes. A much higher variation was even found when p53mut cells were compared with p53null cells with a twofold difference in even 123 genes. The respective genes were found to be involved mainly in apoptosis, cell cycle regulation, metabolisms and signalling but with only one gene relevant for DNA repair. Radiation was found to affect this profile solely for cells with p53wt with a twofold significant up-regulation in only five genes. For selective genes (BCL2, CASP1, CCND2, DDB2, XPC, RAD51C, SESN1, FUCA1, CDKN1A, MDM2, XPC) array data were confirmed by qRT-PCR. Conclusion: The result, that the gene expression profile of lymphoblastoid cells differing in p53 status already displayed clear differences when measured prior to irradiation with only few changes after irradiation, which are solely seen for p53wt cells, suggests, that the differences in radiosensitivity observed for these cells are primarily determined by the variation in expression profile present already prior to irradiation. (c) 2006 Elsevier Ireland Ltd. All rights reserved.
引用
收藏
页码:236 / 249
页数:14
相关论文
共 67 条
[1]   Alterations in gene expression profiles and the DNA-damage response in ionizing radiation-exposed TK6 cells [J].
Akerman, GS ;
Rosenzweig, BA ;
Domon, OE ;
Tsai, CA ;
Bishop, ME ;
McGarrity, LJ ;
MacGregor, JT ;
Sistare, FD ;
Chen, JJ ;
Morris, SM .
ENVIRONMENTAL AND MOLECULAR MUTAGENESIS, 2005, 45 (2-3) :188-205
[2]  
Amundson SA, 2000, RADIAT RES, V154, P342, DOI 10.1667/0033-7587(2000)154[0342:IOPMBI]2.0.CO
[3]  
2
[4]   Stress-specific signatures: expression profiling of p53 wild-type and -null human cells [J].
Amundson, SA ;
Do, KT ;
Vinikoor, L ;
Koch-Paiz, CA ;
Bittner, ML ;
Trent, JM ;
Meltzer, P ;
Fornace, AJ .
ONCOGENE, 2005, 24 (28) :4572-4579
[5]   Fluorescent cDNA microarray hybridization reveals complexity and heterogeneity of cellular genotoxic stress responses [J].
Amundson, SA ;
Bittner, M ;
Chen, YD ;
Trent, J ;
Meltzer, P ;
Fornace, AJ .
ONCOGENE, 1999, 18 (24) :3666-3672
[6]  
Amundson SA, 2001, RADIAT RES, V156, P657, DOI 10.1667/0033-7587(2001)156[0657:IOGEAA]2.0.CO
[7]  
2
[8]   Gene Ontology: tool for the unification of biology [J].
Ashburner, M ;
Ball, CA ;
Blake, JA ;
Botstein, D ;
Butler, H ;
Cherry, JM ;
Davis, AP ;
Dolinski, K ;
Dwight, SS ;
Eppig, JT ;
Harris, MA ;
Hill, DP ;
Issel-Tarver, L ;
Kasarskis, A ;
Lewis, S ;
Matese, JC ;
Richardson, JE ;
Ringwald, M ;
Rubin, GM ;
Sherlock, G .
NATURE GENETICS, 2000, 25 (01) :25-29
[9]   RETRACTED: FOXO1a acts as a selective tumor suppressor in alveolar rhabdomyosarcoma (Retracted Article. See vol 177, pg 563, 2007) [J].
Bois, PRJ ;
Izeradjene, K ;
Houghton, PJ ;
Cleveland, JL ;
Houghton, JA ;
Grosveld, GC .
JOURNAL OF CELL BIOLOGY, 2005, 170 (06) :903-912
[10]   Gene expression signatures identify novel regulatory pathways during murine lung development: implications for lung tumorigenesis [J].
Bonner, AE ;
Lemon, WJ ;
You, M .
JOURNAL OF MEDICAL GENETICS, 2003, 40 (06) :408-417