jlc Journal of Lung Cancer J Lung Cancer 1598-7809 Korean Association for the Study of Lung Cancer 10.6058/jlc.2006.5.2.92 jlc-5-92 Original Article Mutational Analysis of PUMA Gene in Non-small Cell Lung Cancers YooNam JinM.D. LeeJong WooM.S. LeeSung HakM.D. LeeSug HyungM.D. Dartment of Patholoqv. Colleae of Medicine. The Catholic University of Korea, Seoul, Korea Address for correspondence Sug Hyuna Lee, MD. DeDartment of Patholoqv. Colleae of Medicine. The Catholic University of Korea. 505. Banoo-dong, Seochogu, Seoul 137-701. Korea Tel: 82-2-590-1188 Fax; 82-2-537-6586 E-mail; suhulee@catholio.aakr 122006 18122006 5 2 92 95 10112006 01122006 Copyright © 2006 Korean Association for the Study of Lung Cancer 2006 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. Abstract Purpose:

It has become clear that, together with proliferation, deregulation of apoptosis plays a pivotal role in tumorigenesis, and the somatic mutations of apoptosis-related genes have been reported in human cancers. PUMA, a proapoptotic member of Bcl-2 family, mediates p53-deependent and -independent apoptosis. The aim of this study was to explore whether alteration of PUMA protein expression is a characteristic of human lung cancers.

 Materials and Methods:

To explore the possibility that the genetic alterations of PUMA might be involved in the development of human cancers, we analyzed the entire coding region and all splice sites of human PUMA gene in 100 human non-small cell lung cancers (NSCLCs) by polymerase chain reaction (PCR)-based single-strand conformation polymorphism (SSCP),

 Results:

The PCR-SSCP analysis detected no mutation in the entire coding regions and all splice sites of human PUMA gene in the 100 NSCLCs.

 Conclusion:

The data presented here suggested that PUMA gene mutation may not contribute to the pathogenesis of human NSCLCs. (J Lung Cancer 2006;5{2):92-95)

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Representative SSCP of PUMA gene in the non-small cell lung cancers. The exon 2 of the PUMA oene were amplified by PCR using a specific primer set. The PCR products from the representative cases of non-small cell lung cancers were visualized on SSCP. SSCP of DNA from the non-small cell lung cancers (T) shows no aberrant bands as compared to SSCPs from the normal tissues (N),

Sequences of PCR Primer Sets for PUMA Gene Amplification

Pimers Exon Sequence Annealing Tm(°C) Size (bp)
2A 1 F 5’-CTGGTACTTCCCTGCTGACC-3’ 58 172
    R 5’-AGAGGCCGCAGGACACTG-3’    
2B 1 F 5’-CTGCCCGCTGCCTACCTCT-3’ 66 195
    R 5’-CTTCCCGGCTCCTATCACCC-3’    
3 2 F 5’-GGCGGAGCAGCACCTGGAGTCG-3’ 56 187
    R 5’-TCCCACCTGCCGTCTACCCCACC-3’    
4 2 F 5’-CTCAGTCCCCTGCTAACTGC-3’ 56 187
    R 5’-CGAGTCCCTGACGTCCAC-3’