방광암에서 p53유전자의 돌연변이에 대한 연구(2): 동위원소를 사용하지 않는 PCR-SSCP 분석법의 비교분석
p53 Gene Mutations in Bladder Cancer(2): Mulation Analysis by Non-isotopic SSCP Method Sung-Ho Park and Wood-Chul Moon
박성호, 문우철,
소속 상세정보
박성호 ( )
중앙대학교 의과대학 비뇨기과학교실
문우철 ( )
중앙대학교
KMID : 0358319960370080847
Abstract
Badckground.
@EN p53 gene mutations are known to be an important prognostinc factor in bladder cancer. Single strand conformation polymorphism (SSCP) analysis has been suggested to be a promising method to detect p53 gene mutation. However, the technical
pitfalls
associated with conventional SSCP using radioisotope precluded its wide application in clinical practice. We herein have tried non-isotopic SSCP and analyzed the value of this, new method for detecting mutation of p53 gene in bladdeer cancer.
@ES Methods.
@EN In this study of 32 bladder transitional cell carcinoma, we comparatively analyzed polymerase chain reaction (PCR) of exons 5 to 8 of p53 gene, followed by 1) conventional, isotopic-SSCP analysis, 2) non-isotopic SSCP analysis, and 3) DNA
sequencing
analysis.
@ES Results.
@EN On isotopic SSCP analysis, 9 out of 32 cases (28.1%) showed mobility shifts. On non-isotopic SSCP analysis, 10 cases (31.0%) showed mobility shifts. On DNA sequencing analysis, 11 cases (34.3%) showed point mutations. The results of isotopic
SSCP
analysis and non-sotopic SSCP analysis were concordant with that of DNA sequencing in 87.5% and 96.9% of cases, respectively. The sensitivity and specificity of detecting p53 mutations by isotopic and non-isotopic SSCP analysis were estimated to
be
81.8% and 100%, and 91.0% and 100%, respectively. Non-isotopic SSCP significantly reduced the time and cost to analyze p53 mutation to 7.9% and 16.0% of those of isotopic SSCP, respectively (P<0.005).
@ES Conclusions.
@EN Non-isotopic SSCP is a highly sensitive and specific, time-saving, and costeffective method to detect p53 gene mutations in bladder cancer. This new method may be promising for the clinical application.
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