Loss of Heterozygosity of the Retinoblastoma and p53 Genes in Primary Cervical Carcinomas with Human Papillomavirus Infection

Abstract

Objective.Paired DNA samples from 55 primary uterine cervical carcinomas and normal bloods were studied for chromosomal allelic loss (loss of heterozygosity; LOH) of the retinoblastoma (Rb) and p53 gene loci by polymerase chain reaction (PCR) or PCR–restriction fragment length polymorphism analysis. All the study samples contained at least one of the oncogenic human papillomavirus (HPV) type 16 and/or 18 sequences. And the relationships between allelic losses of these genes and conventional clinicopathological parameters were evaluated.Methods.In order to detect LOH of the Rb gene in cervical cancers, we analyzed four polymorphic intronic sites (intron 1, 17, 20, and 25) of the Rb gene and one additional microsatellite near the Rb locus (D13S118). For detection of the LOH in p53, three intragenic polymorphisms (exon 1, exon 4, intron 6) and one microsatellite distal to the p53 gene (D17S5) were examined.Results.By analyzing this system, we could increase the heterozygosity of the Rb and p53 loci up to 0.91 and 1, respectively. The observed allelic loss rates of the Rb and p53 loci in informative cases were 14% (7/50) and 5.5% (3/55), respectively. The patients with LOH at the D13S118 locus also had the allelic loss of the Rb gene, whereas only one of the four patients with LOH at the D17S5 locus showed a concomittant allelic loss of the p53 gene. The frequency of cervical cancer with one LOH at the Rb or p53 loci was 20% (11/55). No shifted bands were observed in the PCR–single-strand conformation polymorphism analysis of the p53 gene. The LOH of the Rb or p53 gene was not significantly associated with other parameters including clinical stage, histological type, degree of differentiation, status of HPV infection, and p53 gene mutation.Conclusion.Concerning the results above, we conclude that the allelic imbalance of the Rb or p53 gene itself is not implicated as a major contributing factor in the malignant transformation or the tumor progression in HPV-positive uterine cervical cancers.