[Analysis on loss of heterozygosity of chromosome 17p13.3 in hepatocellular carcinoma and construction of genomic contig in the deleted region].
OBJECTIVE
To detect the status of loss of heterozygosity (LOH) on chromosome 17p13.3 in hepatocellular carcinoma (HCC) and determine a minimum region of LOH as well as construct genomic contig in LOH region.


METHODS
Variable number tandem repeat (VNTR) and RFLP markers examined by Southern hybridization were used to detect LOH of HCC. Microsatellite markers amplified by PCR were detected for LOH by denatured PAGE. Using microsatellite primers, 3 rounds of PCR amplification were carried out to screen positive genomic clones. The contig was constructed by PCR detecting the reaction between microsatellite markers and individual genomic clone.


RESULTS
Fifty-four paired HCC samples were analysed with 16 polymorphic markers on chromosome 17p13.3. The data revealed that the region between D17S5 and D17S34 had a high rate of LOH (> 63%), but three markers (proximal to D17S5) toward centromere had low or no LOH. TP53 marker on chromosome 17p13.1 had a LOH rate of 31% which was lower than that on D17S5-D17S34 region. No LOH was found at D17S34, D17S1866 (proximal to telomere) and D17S5, D17S1574 (distal to telomere) in 2 cases of HCC. However, there was LOH between D17S849 and D17S1574 in some cases. In LOH region, genomic clones relative to 18 of markers were obtained. A contig covering 9 markers was constructed.


CONCLUSIONS
Determination of the minimum LOH region on chromosome 17p13.3 in HCC and the construction of genomic clone contig provide basis for further identification of putative new tumor suppressor gene(s) in HCC.
Zhao
X
X
National Laboratory for Oncogenes and Related Genes, Shanghai Cancer Institute, Shanghai 200032, China.
Wan
D
D
Jiang
H
H
D006528:Carcinoma, Hepatocellular / Q000235:genetics*; D002874:Chromosome Mapping; D002886:Chromosomes, Human, Pair 17; D006801:Humans; D008113:Liver Neoplasms / Q000235:genetics*; D019656:Loss of Heterozygosity; D018598:Minisatellite Repeats; D012150:Polymorphism, Restriction Fragment Length