Allelic loss determination in chronic lymphocytic leukemia by immunomagnetic bead sorting and microsatellite marker analysis

Abstract

Chronic lymphocytic leukemia (CLL) is the most common adult leukemia in the western world. Details of the molecular mechanisms involved in the pathogenesis of this leukemia are unclear at present. In other malignancies tumorigenesis has been shown to proceed via a multistep progression model with a causal relation between the accumulation of genetic abnormalities and more aggressive clinical behavior. The loss of chromosomal segments in malignant cells has proven useful in mapping regions of DNA that contain candidate tumor suppressor genes. The detection of loss of heterozygosity (LOH) has been greatly facilitated by the analysis of highly polymorphic microsatellite markers enabling the identification of submicroscopic losses. Utilizing immunomagnetic bead sorting to separate leukemic from normal cells we identified at least one allelic losses in 8/29 (28%) of analysed CLL cases with a PCR-based assay. On chromosomal arm 3p we have identified homozygous deletions in 3/29 cases at locus D3S1284 residing in the vicinity of the newly described FHIT gene. Several cases of CLL manifested LOH at a locus telomeric to the p15 and p16 tumor suppressor genes, all being early to intermediate stage disease. Our findings suggest that losses at these loci may contribute to the development and/or progression of CLL in at least a subset of cases.