A PCR-RFLP method for the detection of activated H-ras oncogene with a point mutation at codon 12 and 61.
To investigate the incidence of the H-ras gene activation in bladder tumor and the feasibility of using urinary washout samples for screening, a series of 33 human bladder tumors and their preoperatively collected urinary washout samples were screened using a mutant specific PCR-RFLP (polymerase chain-restriction fragment length polymorphism) to detect a point mutation of the H-ras gene. Five tumors were found to harbor H-ras mutations where two tumors had a glycine to valine (G-->T) change in codon 12 and three tumors had a glutamine to lysine (C-->A) change in codon 61, respectively. Moreover, we could also detect the same point mutations of the H-ras gene in corresponding urine washout samples. The incidence of H-ras mutation in Korean bladder cancer was estimated at approximately 15.2%. In conclusion, a mutant specific PCR-RFLP method for the detection of H-ras gene mutation is useful for screening or postoperative follow-up of bladder tumor due to its simplicity and high specificity even in urinary samples.
Hong
S J
SJ
Department of Urology, Yonsei University College of Medicine, Seoul, Korea.
Lee
T
T
Park
Y S
YS
Lee
K O
KO
Chung
B H
BH
Lee
S H
SH
D001483:Base Sequence; D002295:Carcinoma, Transitional Cell / Q000235:genetics; D003062:Codon; D005786:Gene Expression Regulation; D011905:Genes, ras; D006801:Humans; D008969:Molecular Sequence Data; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D012150:Polymorphism, Restriction Fragment Length; D015533:Transcriptional Activation; D001749:Urinary Bladder Neoplasms / Q000235:genetics
D003062:Codon