ANALYSIS OF CLONALITY BY POLYMERASE CHAIN-REACTION FOR PHOSPHOGLYCERATE KINASE-1 - HETERODUPLEX GENERATOR

Polymerase chain reaction (PCR) amplification has been used to determi ne the clonal composition of tissues based on analysis of the pattern of X-chromosome inactivation, but its use has been limited by technica l difficulties. This report presents an expedited method to use PCR in the analysis of clonality. The method uses gel electrophoresis of het eroduplexes formed with an artificial heteroduplex generator (HG) and PCR products from the phosphoglycerate kinase-1 (PGK-1) gene from the tissue sections. Amplification was successful in 36 of 37 cases origin ally diagnosed as endometrial adenocarcinoma. HG analysis of 36 cases confirmed heterozygosity in 12 cases (33.3%). PGK-1 PCR amplification product was obtained from both control and lesional tissue in 10 of th e 12 heterozygous cases. Of these 10 cases, seven were shown to consis t of clonal cell populations by HG analysis. Two of three cases diagno sed as well-differentiated endometrioid adenocarcinoma were found to b e comprised of polyclonal populations of cells. One case produced an a nomalous pattern with HG analysis and was shown to be aneuploid by flu orescence in situ hybridization (FISH) with a chromosome X alpha-satel lite probe. It is concluded that HG is a useful alternative to restric tion fragment length polymorphism (RFLP) analysis of X-chromosome inac tivation as a marker of tissue clonality in cases in women.