Clonal origin of mammalian hepatitis B virus-related hepatocellular carcinoma

The clonal origins of 20 multifocal hepatocellular carcinomas (HCCs) in four woodchucks were analyzed by the Southern blot hybridization technique. The woodchucks were divided into two groups according to the morphological classification of multifocal tumors: 1) three woodchucks had multifocal tumors that were widely separated and similar in size, which suggests a multiclonal origin of the tumors; and 2) one woodchuck had ten small multifocal tumors surrounding two large main tumors, which indicated intrahepatic metastasis from an original tumor. Results from the first group demonstrated that the number of integrated woodchuck hepatitis virus (WHV) DNAs differed from tumor to tumor, and none of the bands were the same size. In the second group, eight of the ten small tumors surrounding the two large tumors showed the same pattern of WHV DNA integration. One demonstrated an additional band and also shared the same bands with the other tumors, and one small tumor had a different pattern of integration from the others. It was concluded that the clone dissimilarity demonstrated by hybridization patterns does not necessarily mean that HCCs originate independently from different clones, because genetic changes may occur after or at the time of metastasis.