Multiplex PCR screening detects small p53 deletions and insertions in human ovarian cancer cell lines.
Mutations at the p53 tumor suppressor gene locus are a frequent genetic alteration associated with human ovarian carcinoma. Little information exists regarding whether mutational events occur other than point mutations and large deletions, causing loss of heterozygosity. Small intragenic deletions and insertions in the p53 gene have been observed in various human neoplasias. We developed a multiplex polymerase chain reaction (MPCR) screening assay to amplify the complete p53 coding region from genomic DNA in a single step. Deletions and/or insertions were found in six out of 11 newly established ovarian carcinoma cell lines. MPCR detected deletions as small as 2 bp, as confirmed by nucleotide sequence analysis. Most of the observed alterations (6/7) were homozygous or hemizygous. Structural aberrations of the p53 gene possibly leading to loss of p53 cell cycle control may be a consequence of a slipped-mispairing mechanism in rapid DNA replication during repetitious ovulation and wound repair of ovarian epithelial cells. MPCR may be a valuable tool for screening for possible p53 deletion and insertion mutations not only in ovarian cancer but also in other malignancies.
Runnebaum
I B
IB
Molekularbiologie-Labor, Universitäts-Frauenklinik, Ulm, Germany.
Tong
X W
XW
Moebus
V
V
Heilmann
V
V
Kieback
D G
DG
Kreienberg
R
R
D001483:Base Sequence; D004251:DNA Transposable Elements; D004273:DNA, Neoplasm / Q000235:genetics; D005260:Female; D017353:Gene Deletion; D016158:Genes, p53; D006801:Humans; D008969:Molecular Sequence Data; D009838:Oligodeoxyribonucleotides; D010051:Ovarian Neoplasms / Q000235:genetics*; D016133:Polymerase Chain Reaction; D014407:Tumor Cells, Cultured
D004251:DNA Transposable Elements; D004273:DNA, Neoplasm; D009838:Oligodeoxyribonucleotides