[Frontiers In Bioscience, Elite, 10, 375-383, March 1, 2018]

Five- hit hypothesis in ATM gene: An individualized model in a breast cancer patient

Parvin Mehdipour1, Asaad Azarnezhad1

1Department of Medical genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran


1. Abstract
2. Introduction
3. Materials and Methods
3.1. Design of study
3.2. Proband information and clinical data
3.3. DNA extraction, Polymerase chain reaction (PCR), and sequencing
3.4. Molecular cloning
3.5. Immunofluorescence (IF) assay
3.6. In silico analysis
4. Results
4.1. Sequencing findings
4.2. Allelic location of hits
4.3. Immunofluorescence (IF)
4.4. In silico predictions
5. Discussion
6. Acknowledgement
8. References


The aim of this study was to trace D1853N in a proband affected with primary BC to explore the molecular, cellular and structural aspects of ATM. Exon 37 and splicing regions were PCR-sequenced. Allelic location of the alterations was determined by molecular cloning. Possible impact of alterations was investigated through the bioinformatics and protein expression assays. Five genetic variants including IVS 36-91 AA>TT, IVS 36-8 T>C, D1853N, IVS 37+47 A>G, IVS 37+60 Del T were found in the target regions of ATM and all the alterations were occurred heterezygously. IVS 36-8 T>C and D1853N were observed in blood and tumor tissue, whilst splicing variants were only occurred in tumor tissue. Missense D1853N alteration seems to be effective on 2D and 3D structure of ATM protein and the probability of splicing found to be decreased by intronic variants. Protein expression of ATM also confirmed the occurrence and functional impact of alterations. Results reflect a five-hit hypothesis in a proband with BC that influence ATM, as a guard of genomic stability, at molecular, cellular, and structural levels.


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Abbreviations: BC: Breast cancer; ATM: Ataxia telangiectasia-mutated; Family History (FH); Human Splice Finder (HSF); 2D: Secondary structure; 3D: Tertiary structure; IF: Immunofluorescence; PE: Protein expression;

Key Words: Ataxia Telangiectasia Mutated, Polymorphism, Breast cancer, hit- hypothesis, Evolution

Send correspondence to: Parvin Mehdipour, Department of Medical Genetics, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran, Tel: 0098218830100, Fax: 0098218830100, E-mail: mehdipor@tums.ac.ir