Detection of P53 Codon 72 Polymorphism in Lung Cancer Patients by Real Time Polymerase Chain Reaction - SimpleProbe Probe Aassay
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Date
2010-04-07
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Thai Cancer Journal
Abstract
The P53 tumor suppressor gene (P53) is a key gene involved in cancer control by producing P53 protein to inhibit proliferation and enhance apoptosis of cancer cells. It has been reported that the polymorphism at the codon 72 on the exon 4 of P53 resulted in 3 different genotypes of the P53: Arginine/Arginine (Arg/Arg), Arginine/Proline (Arg/Pro) and Proline/Proline (Pro/Pro) genotypes, and found that individual with Pro/Pro of P53 codon 72 polymorphism had a higher risk for lung, bladder and nasopharyngeal cancers than those with Arg/Arg. Polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay used to detect P53 codon 72 polymorphism is not suitable for a mass screening since it is complicated, time-consuming and at risk for carcinogen exposure. Recently, real-time polymerase chain reaction-SimpleProbe probe (R-PCR-SP) assay has been suggested to be suitable for a mass screening of various genetic polymorphisms with a faster and safer way. The aim of this study is to detect P53 codon 72 polymorphism in Thai patients with lung cancer by the R-PCR-SP assay using SimpleProbe probe and melting curve analysis and the PCR-RFLP assay. The findings from these two methods were then comparable. Seventy DNA samples from peripheral blood leukocyte of lung cancer cases were recruited in this study. The results of P53 codon 72 polymorphism detected by the R-PCR-SP and the PCR-RFLP assays were completely accordant (κ=1.0, 95%CI = 1.0-1.0). The findings demonstrated 12 individuals with Arg/Arg, 41 cases with Arg/Pro and 17 cases with Pro/Pro. In addition, we found that the R-PCR-SP assay was a faster and safer method for detection of P53 codon 72 polymorphism than the PCR-RFLP assay. Results of the present study suggest that the R-PCR-SP assay with SimpleProbe probe and melting curve analysis may be a useful screening tool for detection of P53 codon 72 polymorphism in the high risk group of lung cancer in Thais.
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Thai Cancer Journal; Vol.29 No.2 April-June 2009; 63-71