GENOMIC ANALYSIS OF HUMAN HEPATOCELLULAR CARCINOMAS USING RESTRICTIONLANDMARK GENOMIC SCANNING

Restriction Landmark Genomic Scanning (RLGS) was used to examine the m ultiple alterations of genomic DNAs that occur in association with tra nsformation and development of malignancy in primary hepatocellular ca rcinoma (HCC). Genomic DNAs from HCC and its normal counterpart were c leaved by the restriction enzyme NotI, radiolabeled at the cleavage si tes, and then size-fractionated by two-dimensional electrophoresis usi ng HinfI as the second cleavage enzyme. About 2000 spots were recogniz ed, whose position and intensity reflect the locus and the copy number of the corresponding restriction sites. Using this system in combinat ion with micromanipulation of HCC to eliminate possible carry-over of nonmalignant cells, we detected six spots that were decreased in inten sity in common to three different HCCs, along with five that were inte nsified spots. In addition, several spots showed changes that were non overlapping among different tumors.