p53 expression in four human medulloblastoma-derived cell lines.
p53 is a tumor suppressor gene found on the short arm of chromosome 17. Loss of one p53 allele and alteration of the other is found in a variety of tumors, including highgrade glial tumors. Point mutations in the remaining allele occur in a highly conserved region of the gene encompassing exons 5-8. Although 40-50% of medulloblastomas lose sequences on the short arm of chromosome 17, alteration of p53 in these tumors is infrequent. To further characterize genetic alteration of p53 in medulloblastoma, we performed a mutational analysis of four medulloblastoma-derived cell lines established by our laboratory. Using two variable-number tandem repeat markers which map distally to p53, we found evidence indicating loss of sequences on the distal end of chromosome 17p in all four lines. However, no gross alterations of the p53 gene were detected. Northern analysis revealed expression of equivalent amounts of full-length p53 messenger RNA in each cell line. Using the polymerase chain reaction to amplify exons 5-8 of the p53 gene, we directly sequenced the amplified fragments and detected no mutations in any of the cell lines. Our results demonstrate that loss of p53 function through gene deletion and/or recessive mutation is not required for growth in our cell lines.
Srinivasan
J
J
Department of Neurological Surgery, University of Washington Medical Center, Seattle 98195, USA.
Berger
M S
MS
Silber
J R
JR
D000483:Alleles; D002886:Chromosomes, Human, Pair 17; D004249:DNA Damage; D005784:Gene Amplification; D017353:Gene Deletion; D015870:Gene Expression; D016158:Genes, p53 / Q000235:genetics*; D006579:Heterozygote; D006801:Humans; D008527:Medulloblastoma / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger; D014407:Tumor Cells, Cultured
D012333:RNA, Messenger