Abrogation of the p16-Rb pathway in Korean hepatocellular carcinomas.

BACKGROUND/AIMS: The tumor suppressor gene p16 on human chromosome 9p21 encodes a specific inhibitor of the cyclin D-CDK4 complex which inactivates the Rb protein by hyperphosphorylation. Many reports show that p16 is inactivated in a variety of human cancers. We investigated whether abnormalities involving p16 and Rb are associated with hepatocellular carcinomas in Korea.

METHODOLOGY

We performed loss of heterozygosity analysis on 9 primary hepatocellular carcinomas using 7 microsatellite markers spanning human chromosome 9p. Reverse transcriptase-PCR, ribonuclease protection assay, and immunoblotting were used to examine the expression of p16 and Rb in 8 hepatocellular carcinoma cell lines, including 5 from Korean patients. Exons 1 and 2 of the p16 gene were sequenced.

RESULTS

We found a 33% loss of heterozygosity at the D9S171 locus (9p21 region) in the primary hepatocellular carcinomas. The p16 protein was not found in 4 out of 5 (80%) of the Korean cell lines. Among them, 2 cell lines lacked p16 protein and had the following point mutations in p16 exon 2: Asp125 to Asn and Arg58 to Ter. Two of the Korean cell lines and the SK-Hep-1 cells contained deletions in the p16 gene. All cell lines examined, except Hep 3B, expressed Rb protein, which in all cases was dominantly hyperphosphorylated (inactivated).

CONCLUSIONS

Our results suggest that p16 and Rb abnormalities are associated with hepatocellular carcinomas in the Korean population.