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Journal of Zhejiang University SCIENCE B 2007 Vol.8 No.2 P.81~87

http://doi.org/10.1631/jzus.2007.B0081


Screen p53 mutations in hepatocellular carcinoma by FASAY: A novel splicing mutation


Author(s):  WU Xiao-mo, FU Jing-geng, GE Wang-zhong, ZHU Jiang-yan, WANG Jun-yong, ZHANG Wei, QIAN Wei, HUO Ke-ke

Affiliation(s):  Institute of Genetics, School of Life Sciences, Fudan University, Shanghai 200433, China

Corresponding email(s):   kkhuo@fudan.edu.cn

Key Words:  Hepatocellular carcinoma (HCC), p53 mutation, FASAY, Splicing mutation


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WU Xiao-mo, FU Jing-geng, GE Wang-zhong, ZHU Jiang-yan, WANG Jun-yong, ZHANG Wei, QIAN Wei, HUO Ke-ke. Screen p53 mutations in hepatocellular carcinoma by FASAY: A novel splicing mutation[J]. Journal of Zhejiang University Science B, 2007, 8(2): 81~87.

@article{title="Screen p53 mutations in hepatocellular carcinoma by FASAY: A novel splicing mutation",
author="WU Xiao-mo, FU Jing-geng, GE Wang-zhong, ZHU Jiang-yan, WANG Jun-yong, ZHANG Wei, QIAN Wei, HUO Ke-ke",
journal="Journal of Zhejiang University Science B",
volume="8",
number="2",
pages="81~87",
year="2007",
publisher="Zhejiang University Press & Springer",
doi="10.1631/jzus.2007.B0081"
}

%0 Journal Article
%T Screen p53 mutations in hepatocellular carcinoma by FASAY: A novel splicing mutation
%A WU Xiao-mo
%A FU Jing-geng
%A GE Wang-zhong
%A ZHU Jiang-yan
%A WANG Jun-yong
%A ZHANG Wei
%A QIAN Wei
%A HUO Ke-ke
%J Journal of Zhejiang University SCIENCE B
%V 8
%N 2
%P 81~87
%@ 1673-1581
%D 2007
%I Zhejiang University Press & Springer
%DOI 10.1631/jzus.2007.B0081

TY - JOUR
T1 - Screen p53 mutations in hepatocellular carcinoma by FASAY: A novel splicing mutation
A1 - WU Xiao-mo
A1 - FU Jing-geng
A1 - GE Wang-zhong
A1 - ZHU Jiang-yan
A1 - WANG Jun-yong
A1 - ZHANG Wei
A1 - QIAN Wei
A1 - HUO Ke-ke
J0 - Journal of Zhejiang University Science B
VL - 8
IS - 2
SP - 81
EP - 87
%@ 1673-1581
Y1 - 2007
PB - Zhejiang University Press & Springer
ER -
DOI - 10.1631/jzus.2007.B0081


Abstract: 
Objective: To establish a routine procedure for the detection of p53 mutations in hepatocellular carcinoma (HCC) surgical resections using the FASAY (functional analysis of separated alleles of p53 on yeast) procedure. Methods: p53 status was analyzed by FASAY and cDNA sequencing in 50 cases of HCC. After the extraction of RNA from the frozen tumor and corresponding normal tissues, reverse transcription RT-PCR was carried out using these samples. The assay can detect mutations of p53 mRNA between codons 67 and 347 by the DNA-binding activity of the protein and reveal them as red colonies. Results: Of the 50 specimens, 29 (58%) were positive (mutant) by FASAY. Sequencing analysis confirmed that all 29 FASAY positive tumors harbored mutations, and that no mutations were detectable in any FASAY negative tumors. In 29 p53 mutations, 22 mutations were point missense mutation, 5 were deletions and 2 were splicing mutations. A novel splice mutation on splice donor of intron 6 was reported, which could produce two different mRNAs, respectively using the nearest upstream and downstream recessive splice donor sites. Conclusion: FASAY is a sensitive method for detecting the various types of p53 mutations in HCC, suggesting that the yeast functional assay for the detection of p53 mutations may be essential for elucidating their clinical significance.

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