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Identification of two genetic alterations in oral premalignancies and tumors using a novel fingerprinting assay Ishkanian, Adrian Shea

Abstract

The identification of novel markers causal in oral tumorigenesis is critical for clinical diagnosis and intervention. The study of oral premalignancies is necessary in order to identify these early stage genetic events. As such tissues are typically available as archival formalin-fixed paraffin-embedded biopsies, we have modified the standard RAPD (Randomly Amplified Polymorphic DNA) fingerprinting technique to utilize this material. This modified assay SMAL (Scanning Minute Archival Lesions) enables reproducible high-density fingerprinting of specific cell populations dissected either manually or through LCM (Laser Capture Microdissection). 235 paraffin-embedded formalin-fixed oral tumor and premalignant biopsies were tested using the SMAL assay. Two novel genetic changes in oral cancer progression stages were identified. These changes were mapped to chromosomes 2p23 and 7q22. Genetic instability at these two loci was verified by LOH (Loss Of Heterozygosity) analysis. 72 oral specimens (38 tumors and 34 dysplasias) were used to define a minimal boundary of allelic imbalance at these loci by LOH. Candidate genes Anaplastic Lymphoma Kinase, Dynein, Phosphatidylinositol 3-Kinase and Fos-Like Antigen 2 have been identified.

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