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12 March 2002

Loss of heterozygosity in primary lung cancer using laser capture microdissection and WAVE DNA fragment analysis techniques.

Helen Zou, Amy Beck, Dave Chervinsky, Weigang Zhu, Debbie Malik, John Brooks, Dongfeng Tan

Med Sci Monit 2002; 8(3): BR95-99 :: ID: 420892

Abstract

BACKGROUND:A number of molecular changes observed by varied conventionalmethods, including loss of heterozygosity (LOH) on chromosome 3, have been associated with primary lungcancer. To further define the locus of chromosome 3p allele loss in lung cancer, we performed LOH studyby using innovative laser capture microdissection and WAVE DNA Fragment Analysis. MATERIAL/METHODS: Thirty-eightpaired specimens from patients with adenocarcinoma of the lung were used for this study. Formalin-fixed,paraffin-embedded tissue from normal stromal cells or lymphocytes and adenocarcinoma were collected usinglaser capture microdissection. DNA was extracted and amplified by PCR using six polymorphic DNA markersfor chromosome 3. PCR products were analyzed by both gel electrophoresis and WAVE DNA Fragment Analysis.RESULTS: LOH at 3p22-24 was found in tumor cells from twelve out of thirty-eight patients (32%) whenanalyzed by WAVE DNA Fragment Analysis and LOH was found in tumor cells from nine out of thirty-eightpatients (23%) when analyzed by gel electrophoresis. LOH was found in normal control from one out ofthirty-eight patients. CONCLUSIONS: 1. Our results suggest putative tumor suppressor gene(s) is presentin a region at 3p22-24, which may play a role in carcinogenesis of lung cancer. 2. Laser capture microdissectionis essential tool for defined LOH studies. 3. WAVE DNA Fragment Analysis is an accurate, sensitive andautomated tool for analysis of DNA fragments.

Keywords: Adenocarcinoma, Aged, 80 and over, Alleles, Chromosomes, Human, Pair 3, DNA Fragmentation, Lasers, Loss of Heterozygosity, Lung Neoplasms, Polymerase Chain Reaction, Polymorphism, Genetic, Research Support, Non-U.S. Gov

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Medical Science Monitor eISSN: 1643-3750
Medical Science Monitor eISSN: 1643-3750