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Detection of p53 Gene Mutations by Single Strand Conformation Polymorphisms Analysis in Various Human Liver Cancer Cell Lines Using a Non-radioactive Mini Gel Electrophoresis System

Khairul Bariah Ahmad Amin Noordin, and A. Rahman A. Jamal, and Wan Ngah W.Z., (2008) Detection of p53 Gene Mutations by Single Strand Conformation Polymorphisms Analysis in Various Human Liver Cancer Cell Lines Using a Non-radioactive Mini Gel Electrophoresis System. Malaysian Journal of Biochemistry and Molecular Biology, 16 (1). pp. 22-26. ISSN ISSN 1511-2616

Full text not available from this repository.

Official URL: http://ejum.fsktm.um.edu.my/ArticleInformation.aspx?ArticleID=600

Affiliations

Hospital Universiti Kebangsaan Malaysia. UKM Medical Molecular Biology Institute
Universiti Kebangsaan Malaysia. Faculty of Medicine, Dept. of Paediatrics
Universiti Kebangsaan Malaysia. Faculty of Medicine, Dept. of Biochemistry

Abstract

Previous studies have reported that human liver cancer cells derived from patients with hepatocellular carcinoma (HCC), for example, Alexander (PLC/PRF/5), have p53 gene mutations in different codons. Detection of these mutations by single strand conformation polymorphisms (SSCP) has been reported useful in p53 mutation studies in HCC. The aim of this study was to determine whether SSCP analysis using a mini gel electrophoresis system, followed by visualisation of electrophoretic band using silver staining was able to detect the p53 gene mutations. Exons 3, 5, 6, 7, 8, 9, 10 and 11 of the p53 gene of five human HCC cell lines (Alexander, HLE, HLF, HuH7 and HepG2) and two normal liver cell lines (Chang and WRL-68) were amplified using polymerase chain reaction (PCR), and the PCR products were later subjected to SSCP analysis. Our results showed that the migration patterns for exon 7 in the HCC cell lines, Alexander, HLE and HLF, were different from those of the normal cell lines, Chang and WRL-68. However, the cell lines HepG2 and HuH7 demonstrated the same migration patterns as those of the two normal cell lines. For exon 6, all of the cell lines except HuH7, showed the same mobility shifts with the presence of a band. For exons 3, 5, 8, 9, 10 and 11, the migration patterns for all HCC cell lines were as those of the normal cell lines. Further examination by direct sequencing confirmed the presence of mutations in the respective HCC cell lines in exons 6 and 7 of the p53 gene. In conclusion, p53 gene mutations could be detected and differentiated using our SSCP protocol.

Item Type:Journal
Keywords:p53 gene mutations, hepatoocellular carcinoma cell lines, SSCP
Subjects:Q Science, Computer Science
R Medicine, Dentistry, Pharmacy, Nursing
ID Code:2170

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