Paraffin wax-embedded material as a source of DNA for the detection of somatic genetic changes.
Loss of genetic material, corresponding to chromosomal deletions, has been detected in a wide range of tumours and may indicate the position of a tumour suppressor gene. In order to identify the position of such a gene more precisely, many tumour samples must be studied until a minimum consensus deletion is characterized. This process is particularly necessary for lung tumours in which the deletion in chromosome 3, seen with such high frequencies in all histological subtypes, is almost always large. We have recently described the use of the polymerase chain reaction (PCR) for restriction fragment length polymorphism (RFLP) analysis of DNA isolated from small bronchial biopsies of lung tumours. In this study we adapted this technique to allow genotyping of DNA isolated from paraffin wax-embedded material (PWEM) microdissected from glass slides. We have investigated 12 lung tumours at polymorphic loci on chromosome 3 and showed allelic loss in all samples. In adapting PCR-RFLP analysis for DNA isolated from PWEM, we have concentrated on those approaches which might be adaptable to routine clinical practice. Somatic genetic changes are now being identified in many tumour types, and this information is expected to be of diagnostic and prognostic significance.
Sundaresan
V
V
Medical Research Council, Clinical Oncology and Radiotherapeutics Unit, Cambridge, U.K.
Ganly
P
P
Hasleton
P
P
Bleehen
N M
NM
Rabbitts
P
P
D000483:Alleles; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics; D002872:Chromosome Deletion; D002893:Chromosomes, Human, Pair 3; D004273:DNA, Neoplasm / Q000032:analysis*; D005838:Genotype; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D016612:Paraffin Embedding; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length
D004273:DNA, Neoplasm