Mutation of the tumor suppressor gene p53 in human prostate and bladder cancers--investigation by temperature gradient gel electrophoresis (TGGE).
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Mutation of the tumor suppressor gene p53 in human prostate and bladder cancers--investigation by temperature gradient gel electrophoresis (TGGE). / Schlechte, H H; Schnorr, D; Löning, Thomas; Rudolph, B D; Pohrt, U M; Loening, S A.
in: J UROLOGY, Jahrgang 157, Nr. 3, 3, 1997, S. 1049-1053.Publikationen: SCORING: Beitrag in Fachzeitschrift/Zeitung › SCORING: Zeitschriftenaufsatz › Forschung › Begutachtung
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TY - JOUR
T1 - Mutation of the tumor suppressor gene p53 in human prostate and bladder cancers--investigation by temperature gradient gel electrophoresis (TGGE).
AU - Schlechte, H H
AU - Schnorr, D
AU - Löning, Thomas
AU - Rudolph, B D
AU - Pohrt, U M
AU - Loening, S A
PY - 1997
Y1 - 1997
N2 - PURPOSE: To test the value of a recently developed screening method for the detection of p53 mutations in prostate and bladder cancers. MATERIALS AND METHODS: Tumor tissue from 24 prostate cancers and 27 bladder cancers were evaluated. DNA of the critical p53 exons 5-8 were amplified and run on horizontal polyacrylamide gels under defined temperature conditions (TGGE) to yield specific gel shifts and sets of homo- and heteroduplexes in case of mutation. Sequencing with a laser-fluorescent electrophoresis unit was done directly from polymerase chain reaction (PCR) products and/or from reamplified mutant and wild type bands excised from the gels. RESULTS: The p53 genotype predicted from the TGGE analysis was always confirmed on the excised DNA fragments, in contrast to only 50% of cases tested by direct sequencing from mixed wild type and mutant DNA present in PCR products. With this screening protocol, 6 of 24 prostate cancers (25.0%) and 11 of 27 bladder cancers (40.7%) showed p53 mutations. At stage T1, none of prostate cancers and 41.2% of bladder cancers contained mutant p53. At higher stages (> or = T2), 30.0% of prostate cancer and 50.0% of bladder cancers were mutated. Histological tumor grading was > G1 in all but two prostate/bladder cancers with mutant p53. It appears that p53 mutations can occur early in bladder carcinogenesis. CONCLUSION: TGGE fulfills the clinical need of a rapid and specific screening method, and, at the molecular level, has the advantage of sorting out the wild type and mutant alleles for consecutive sequencing.
AB - PURPOSE: To test the value of a recently developed screening method for the detection of p53 mutations in prostate and bladder cancers. MATERIALS AND METHODS: Tumor tissue from 24 prostate cancers and 27 bladder cancers were evaluated. DNA of the critical p53 exons 5-8 were amplified and run on horizontal polyacrylamide gels under defined temperature conditions (TGGE) to yield specific gel shifts and sets of homo- and heteroduplexes in case of mutation. Sequencing with a laser-fluorescent electrophoresis unit was done directly from polymerase chain reaction (PCR) products and/or from reamplified mutant and wild type bands excised from the gels. RESULTS: The p53 genotype predicted from the TGGE analysis was always confirmed on the excised DNA fragments, in contrast to only 50% of cases tested by direct sequencing from mixed wild type and mutant DNA present in PCR products. With this screening protocol, 6 of 24 prostate cancers (25.0%) and 11 of 27 bladder cancers (40.7%) showed p53 mutations. At stage T1, none of prostate cancers and 41.2% of bladder cancers contained mutant p53. At higher stages (> or = T2), 30.0% of prostate cancer and 50.0% of bladder cancers were mutated. Histological tumor grading was > G1 in all but two prostate/bladder cancers with mutant p53. It appears that p53 mutations can occur early in bladder carcinogenesis. CONCLUSION: TGGE fulfills the clinical need of a rapid and specific screening method, and, at the molecular level, has the advantage of sorting out the wild type and mutant alleles for consecutive sequencing.
M3 - SCORING: Zeitschriftenaufsatz
VL - 157
SP - 1049
EP - 1053
JO - J UROLOGY
JF - J UROLOGY
SN - 0022-5347
IS - 3
M1 - 3
ER -