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FREQUENCY OF H-RAS MUTATIONS IN HUMAN BLADDER-CANCER DETECTED BY DIRECT SEQUENCING

Frequency of H-ras mutations in human bladder cancer detected by direct sequencing

Lookup NU author(s): Professor David Neal, Professor John LunecORCiD

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Abstract

Objective To determine the frequency of mutations of the H-ras gene in transitional cell carcinomas of the human urinary bladder using direct DNA sequencing based on the polymerase chain reaction (PCR) method and to compare the results with those of other methods. In addition, the relationship of the mutation frequency to tumour stage and grade was examined. Patients and methods Bladder tumour samples, taken by cystoscopic resection from 50 patients with newly diagnosed transitional cell carcinoma of the urinary bladder, were analysed by PCR-based direct DNA sequencing for point mutations in the H-ras gene at codon 12. Results Point mutations were found in 9 of 50 tumours examined (18%). The most frequent mutation (8/9) was a G to T transversion converting GGC to GTC, which would result in a glycine to valine substitution. The remaining mutation was a G to A transition altering GGC to GAC, producing a glycine to aspartic acid substitution, which has not previously been reported in bladder cancer. In all tumour samples examined the wild-type allele (GGC) was also evident. Variation in the proportion of wild-type to mutated sequence was found within tumour samples. No relationship between mutations and tumour grade and stage was apparent. Conclusion The frequency of H-ras mutations detected in this first large scale study using the highly sensitive and rapid PCR-based sequencing method was comparable to that reported by earlier studies with the nude mouse tumorigenesis variation of the 3T3 transfection assay. H-ras mutations can be early events in the development and progression of a significant proportion of human bladder cancer cases.

Publication metadata

Author(s): Burchill, S.A., Neal, D.E., Lunec, J.

Publication type: Article

Publication status: Unknown

Journal: British Journal of Urology

Year: 1994

Volume: 73

Issue: 5

Pages: 516-521

Print publication date: 01/05/1994

ISSN (print): 1464-4096

ISSN (electronic): 1464-410X

URL: http://dx.doi.org/10.1111/j.1464-410X.1994.tb07636.x

DOI: 10.1111/j.1464-410X.1994.tb07636.x

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