This Article Full Text Full Text (PDF) Alert me when this article is cited Alert me if a correction is posted Services Similar articles in this journal Similar articles in PubMed Alert me to new issues of the journal Download to citation manager Citing Articles Citing Articles via HighWire Citing Articles via Google Scholar Google Scholar Articles by Qian, G.-S. Articles by Jackson, P. E. Search for Related Content PubMed PubMed Citation Articles by Qian, G.-S. Articles by Jackson, P. E.

Sensitivity of Electrospray Ionization Mass Spectrometry Detection of Codon 249 Mutations in the p53 Gene Compared with RFLP¹

Department of Environmental Health Sciences, Johns Hopkins Bloomberg School of Public Health, Baltimore, Maryland 21205 [S-Y. K., X. H., J. D. G., P. E. J.], and Shanghai Cancer Institute, Shanghai 200032, People’s Republic of China [G-S. Q.]

Hepatocellular carcinoma (HCC) has several major etiological risk factors, including infection with hepatitis viruses and exposure to aflatoxin B₁. A specific missense mutation resulting from a guanine to thymine transversion at the third position of codon 249 in the p53 tumor suppressor gene has been reported in 10–70% of HCCs from areas of high dietary exposure to aflatoxin B_1. This mutation has not only been detected in tumor samples but has also been measured in DNA isolated from the blood of patients with HCC in two separate studies by two independent methods: RFLP and short oligonucleotide mass analysis (SOMA), an electrospray ionization mass spectrometry technique. To compare the relative sensitivities of these methodologies, a set of serially diluted samples was analyzed by both techniques. The detection limits of RFLP and SOMA were 6% and 2.4% mutant alleles in the presence of wild-type alleles, respectively. When the DNA samples were predigested with HaeIII before SOMA, the detection limit was improved to 0.4% mutant allele in the presence of wild-type alleles. We have therefore found that SOMA is about 2.5–15-fold more sensitive than RFLP for detection of specific p53 mutations. A set of 26 DNA samples from HCC and normal liver was analyzed by RFLP and SOMA, and 5 samples were positive for the p53 mutation. An additional 4 samples were found to be positive for the mutation when SOMA was repeated after HaeIII predigestion.

This article has been cited by other articles:

				R. Singh and P. B. Farmer Liquid chromatography-electrospray ionization-mass spectrometry: the future of DNA adduct detection Carcinogenesis, February 1, 2006; 27(2): 178 - 196. [Abstract] [Full Text] [PDF]

				M. E. Lleonart, G. D. Kirk, S. Villar, O. A. Lesi, A. Dasgupta, J. J. Goedert, M. Mendy, M. C. Hollstein, R. Montesano, J. D. Groopman, et al. Quantitative Analysis of Plasma TP53 249Ser-Mutated DNA by Electrospray Ionization Mass Spectrometry Cancer Epidemiol. Biomarkers Prev., December 1, 2005; 14(12): 2956 - 2962. [Abstract] [Full Text] [PDF]

				M. E. Lleonart, S. Ramon y Cajal, J. D. Groopman, and M. D. Friesen Sensitive and specific detection of K-ras mutations in colon tumors by short oligonucleotide mass analysis Nucleic Acids Res., March 22, 2004; 32(5): e53 - e53. [Abstract] [Full Text] [PDF]

				P. E. Jackson, S.-Y. Kuang, J.-B. Wang, P. T. Strickland, A. Munoz, T. W. Kensler, G.-S. Qian, and J. D. Groopman Prospective detection of codon 249 mutations in plasma of hepatocellular carcinoma patients Carcinogenesis, October 1, 2003; 24(10): 1657 - 1663. [Abstract] [Full Text] [PDF]