Screening of human bladder tumors and urine sediments for the presence of H-ras mutations.
A series of 111 human bladder tumors were screened using oligonucleotide mutant specific probes, restriction enzyme analysis and single-stranded confirmation polymorphism (SSCP) for the presence of H-ras activation events. Thirty-three tumors were found to harbor H-ras mutations where a glycine to valine (G-->T) change in codon 12 was the most common point mutation recorded (26 tumors). Additional mutations involved glycine to cysteine at codon 13 (2 tumors) and glutamine to arginine/lysine/leucine at codon 61 (3/1/1 tumors, respectively). Ambiguous signals recorded with oligonucleotide probes were further analyzed using SSCP analysis revealing the presence of H-ras mutations in restricted regions of some tumors. The apparent sensitivity of SSCP enabled us to extend this study to DNA isolated from urine sediments where 4 of the 9 patients studied showed representation of mutant H-ras. Our study demonstrates a sensitive, non-invasive assay for the screening of urine-borne cells, with no requirement for prior knowledge of the mutational change at the H-ras locus.
Levesque
P
P
Department of Surgery, New England Deaconess Hospital, Harvard Medical School, Boston, MA 02215.
Ramchurren
N
N
Saini
K
K
Joyce
A
A
Libertino
J
J
Summerhayes
I C
IC
D016475:3T3 Cells; D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002460:Cell Line; D004273:DNA, Neoplasm / Q000032:analysis* / Q000652:urine; D004277:DNA, Single-Stranded; D011905:Genes, ras; D006801:Humans; D051379:Mice; D008969:Molecular Sequence Data; D009154:Mutation; D015345:Oligonucleotide Probes; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D014162:Transfection; D001749:Urinary Bladder Neoplasms / Q000235:genetics*
D004273:DNA, Neoplasm; D004277:DNA, Single-Stranded; D015345:Oligonucleotide Probes