Genomic analysis of human hepatocellular carcinomas using Restriction Landmark Genomic Scanning

Restriction Landmark Genomic Scanning (RLGS) was used to examine the multiple alterations of genomic DNAs that occur in association with transformation and development of malignancy in primary hepatocellular carcinoma (HCC). Genomic DNAs from HCC and its normal counterpart were cleaved by the restriction enzyme NotI, radiolabeled at the cleavage sites, and then size-fractionated by two-dimensional electrophoresis using HinfI as the second cleavage enzyme. About 2000 spots were recognized, whose position and intensity reflect the locus and the copy number of the corresponding restriction sites. Using this system in combination with micromanipulation of HCC to eliminate possible carry-over of nonmalignant cells, we detected six spots that were decreased in intensity in common to three different HCCs, along with five that were intensified spots. In addition, several spots showed changes that were nonoverlapping among different tumors.