Mutation analysis of p53 in ovarian rumors by DHPLC

Up to now, ovarian carcinomas represent a major health problem among female cancers because they are the leading cause of death from gynecological mal ignancy. A high proportion of these tumors selects for mutations in the p53 gene. There is evidence that inactivation of the p53 protein could indicat e poor prognosis and chemoresistance of patients. To set up a fast and sens itive test for p53 defects in tumor tissues. we analyzed ovarian cancer cel ls by denaturing high-performance liquid chromatography (DHPLC). A primer s et spanning the whole coding region of p53 with seven fragments was designe d and appropriate heteroduplex detection in DHPLC analysis was elaborated. The analysis of 45 ovarian tumor specimens yielded 17 genetic alterations ( 38%) occurring exclusively in the malignant tissue of the patients. In addi tion, frequent polymorphisms present in normal compared to tumor tissue cou ld serve as a tool for the rapid identification of loss of heterozygosity ( LOH) in the tumor. We observed that LOH in intron 2 or 3 correlated well wi th a lack of one allele in mutated fragments. In conclusion, DHPLC screenin g appears to be a sensitive and effective test for genetic alterations in t umors with p53 involvement. Since p53 mutations point to a poor prognosis s tate in several cancers. a fast screening of turner material for genetic va riations may have implications for further individual treatment of patients . (C) 2001 Elsevier Science B.V. All rights reserved.