Lab Med Online LMO Laboratory Medicine Online 2093-6338 The Korean Society for Laboratory Medicine 10.3343/lmo.2011.1.4.6 Original Article Diagnostic Genetics Analysis of TP53 Gene Mutations in the Korean Patients with Lung Cancer Jung Sang Bong MS 1 Chang Hee Kyung M.D. 1 Choi Seok Cheol Ph.D. 2 Shin Kyeong-Cheol M.D. 3 Lee Kwan-Ho M.D. 3 Jeon Chang Ho M.D. 4 Department of Pathology, Kosin University College of Medicine, Busan, Korea. Department of Clinical Laboratory Science, Catholic University, Busan, Korea. Department of Internal Medicine, College of Medicine, Yeungnam University, Daegu, Korea. Department of Laboratory Medicine, Daegu Catholic Medical Center, Daegu, Korea. Corresponding author: Chang Ho Jeon, M.D. Department of Laboratory Medicine, Daegu Catholic Medical Center, 3056-6 Daemyung 4-dong, Nam-gu, Daegu 705-718, Korea. Tel: +82-53-650-4144, Fax: +82-53-653-8672, chjeon@cu.ac.kr 10 2011 07 10 2011 1 4 202 208 05 03 2011 21 04 2011 30 05 2011 © 2011, Laboratory Medicine Online 2011

This is an Open Access article distributed under the terms of the Creative Commons Attribution Non-Commercial License (http://creativecommons.org/licenses/by-nc/3.0/) which permits unrestricted non-commercial use, distribution, and reproduction in any medium, provided the original work is properly cited.

 Background

Lung cancer is one of the leading causes of cancer-related deaths throughout the world. The gene tumor protein 53 (TP53) is frequently mutated in cases of lung cancer. This study was performed to investigate the frequencies and types of mutations in the TP53 gene in Korean patients with lung cancer.

 Methods

We obtained tissue samples from 80 lung cancer patients and synthesized TP53 cDNA by using RNA isolated from these tissues by performing reverse transcriptase polymerase chain reaction. Hybridization and denaturing high-performance liquid chromatography were performed to identify the TP53 gene mutations, and then, the mutations were validated by direct sequencing.

 Results

Forty mutations out of the 80 patients (50.0%) were noted in the TP53 gene. The frequencies of TP53 gene mutation for different cancer types, namely, squamous cell carcinoma, adenocarcinoma, and small cell carcinoma were 61.1%, 27.3%, and 26.7%, respectively. The mutation frequencies in the different regions of the gene were 10.0% for exon 4, 35.0% for exon 5, 12.5% for exon 6, 22.5% for exon 7, 17.5% for exon 8, and 2.5% for exon 9. The frequently mutated positions were codon 179 in exon 5, codons 202 and 220 in exon 6, and codons 266 and 273 in exon 8.

 Conclusions

Exon 5 was the most frequently mutated region in the TP53 gene. Compared to the patients with the other types of cancers, patients with squamous cell carcinoma showed a higher frequency of TP53 mutation. Codon 179 was the most frequently mutated codon in the TP53 gene.

 TP53 gene mutation denaturing high performance liquid chromatography exon codon

This article is available from http://www.labmedonline.org

 Minna JD Pass HI Mitchell JB Johnson DH Turrisi AT Molecular biology overview Lung cancer: principles and practice 1996 1st ed Philadelphia Lippincott-Raven 143 148 Mendelsohn J Targeting the epidermal growth factor receptor for cancer therapy J Clin Oncol 2002 20 18 Suppl 1S 13S Sozzi G Pastorino U Moiraghi L Tagliabue E Pezzella F Ghirelli C Loss of FHIT function in lung cancer and preinvasive bronchial lesions Cancer Res 1998 58 5032 5037 Mao L Recent advances in the molecular diagnosis of lung cancer Oncogene 2002 21 6960 6969 Olivier M Hollstein M Hainaut P TP53 mutations in human cancers: origins, consequences, and clinical use Cold Spring Harb Perspect Biol 2010 2 a001008 Hainaut P Hollstein M p53 and human cancer: the first ten thousand mutations Adv Cancer Res 2000 77 81 137 Vogelstein B Lane D Levine AJ Surfing the p53 network Nature 2000 408 307 310 Pfeifer GP Besaratinia A Mutational spectra of human cancer Hum Genet 2009 125 493 506 Jassem E Nikliński J Rosell R Niklińska W Jakóbkiewicz J Monzo M Types and localisation of p53 gene mutations: a report on 332 non-small cell lung cancer patients Lung Cancer 2001 34 Suppl 2 S47 S51 Feng Z Hu W Rom WN Costa M Tang MS Chromium(VI) exposure enhances polycyclic aromatic hydrocarbon-DNA binding at the p53 gene in human lung cells Carcinogenesis 2003 24 771 778 Pfeifer GP Denissenko MF Oliver M Tretyakova N Hecht SS Hainaut P Tobacco smoke carcinogens, DNA damage and p53 mutations in smoking-associated cancers Oncogene 2002 21 7435 7451 Hussain SP Amstad P Raja K Sawyer M Hofseth L Shields PG Mutability of p53 hotspot codons to benzo(a)pyrene diol epoxide (BPDE) and the frequency of p53 mutations in nontumorous human lung Cancer Res 2001 61 6350 6355 Lee EB Jin G Lee SY Park JY Kim MJ Choi JE TP53 mutations in Korean patients with non-small cell lung cancer J Korean Med Sci 2010 25 698 705 Walker DR Bond JP Tarone RE Harris CC Makalowski W Boguski MS Evolutionary conservation and somatic mutation hotspot maps of p53: correlation with p53 protein structural and functional features Oncogene 1999 18 211 218 Kato S Han SY Liu W Otsuka K Shibata H Kanamaru R Understanding the function-structure and function-mutation relationships of p53 tumor suppressor protein by high-resolution missense mutation analysis Proc Natl Acad Sci USA 2003 100 8424 8429 Takahashi T Nau MM Chiba I Birrer MJ Rosenberg RK Vinocour M p53: a frequent target for genetic abnormalities in lung cancer Science 1989 246 491 494 Hu Y McDermott MP Ahrendt SA The p53 codon 72 proline allele is associated with p53 gene mutations in non-small cell lung cancer Clin Cancer Res 2005 11 2502 2509 Sidransky D Hollstein M Clinical implications of the p53 gene Annu Rev Med 1996 47 285 301 Horio Y Takahashi T Kuroshi T Hibi K Suyama M Niimi T Prognostic significance of p53 mutations and 3p deletions in primary resected non-small cell lung cancer Cancer Res 1993 53 1 4 Hashimoto T Tokuchi Y Hayashi M Kobayashi Y Nishida K Hayashi S Different subtypes of human lung adenocarcinoma caused by different etiological factors. Evidence from p53 mutational spectra Am J Pathol 2000 157 2133 2141 International Agency for Research on Cancer. IARC tp53 database, R15 World Health Organization Updated on Nov 2010 http://www-p53.iarc.fr/index.html Hernandez-Boussard TM Hainaut P A specific spectrum of p53 mutations in lung cancer from smokers: review of mutations compiled in the IARC p53 database Environ Health Perspect 1998 106 385 391 Bennett WP Hussain SP Vahakangas KH Khan MA Shields PG Harris CC Molecular epidemiology of human cancer risk: gene-environment interactions and p53 mutation spectrum in human lung cancer J Pathol 1999 187 8 18 Pfeifer GP Denissenko MF Tang M p53 mutations, benzo[a]pyrene and lung cancer: a reply Mutagenesis 1998 13 537 538

Denaturing high-performance liquid chromatography (DHPLC) analysis of exons 4-5 and 6-9.

Chromatograms obtained by performing DHPLC for exons 4-5 (A) and 6-9 (B) of wild type (a) and mutated (b) TP53 gene. The chromatograms for both the regions of the wild type gene were similar to those for the corresponding regions of the controls, while those for both the regions of the mutant genes were different from those for the corresponding regions of the controls.

Characteristics of the lung cancer patients

Abbreviations: Sqcc, squamous cell carcinoma; Smcc, small cell carcinoma; Adeno, adenocarcinoma.

Primer sequences used for detecting mutations in exons 4 to 9 of the TP53 gene

Abbreviation: bp, base pairs.

TP53 mutation frequencies for the different types of lung carcinomas

*One patient had a compound mutation at exon 6 and 8.

Types of TP53 mutations in Korean patients with lung cancer

*Novel mutation.

Abbreviations: Adeno, adenocarcinoma; Smcc, small cell carcinoma; Sqcc, squamous cell carcinoma; FS, frameshift; M, male; F, female; c, codon; p, protein.

Comparison of our results with those of previous studies on lung cancer

Abbreviation: NT, not tested.