Detection of DNA abnormalities by arbitrarily primed PCR fingerprinting: Amplification of the MDM2 gene in a mediastinum fibrosarcoma

Arbitrarily primed PCR (AP-PCR) fingerprinting method is easy and useful fo r analysis of genetic alterations in anonymous chromosomal regions. We appl ied this technology to analysis of DNA from human primary cancers and found amplification of a DNA fragment in a mediastinum fibrosarcoma. PCR-based a nalysis of radiation hybrid panels following cloning and nucleotide sequenc e determination of the fragment revealed that it was derived from a region of chromosome 12q13-q15. In this region, the MDM2 and IFNG genes were noted as known genes that could be involved in human carcinogenesis. Southern bl ot analysis of genomic DNA of the tumor revealed the amplification of the M DM2 gene together with the fragment locus, but not the IFNG gene. Our resul ts demonstrated that detection of DNA alterations by AP-PCR fingerprinting without any previous knowledge of the genes and subsequent analysis of radi ation hybrid panels could lead to easy identification of candidates for gen es involved in carcinogenesis. (C) 1999 Academic Press.