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2006 | 09 | 2 |
Tytuł artykułu

Real-Time PCR analysis of p53 mRNA levels in tissues of whitefish [Coregonus lavaretus] exposed to benzo[a]pyrene

Warianty tytułu
Języki publikacji
EN
Abstrakty
EN
We developed a real-time PCR assay for measuring relative quantities (RQ) of p53 tumor suppressor mRNA in the whitefish (Coregonus lavaretus, Salmonidae, Teleostei). Real-time PCR primers for the p53 gene were designed from a region that was found to be conserved among salmonid p53 genes. To test for the usefulness of the assay we performed a treatment study, using benzo[a]pyrene (B[a]P) a putative p53-inducer. Two groups of hatchery raised whitefish, with an average body mass of 15 g and total length of 12 cm were either given an intraperitoneal injection (10 mg • kg-1) of B[a]P in corn oil (2 mg B[a]P ml-1 corn oil) or corn oil alone (Control). After treatment (48 h, 7°C), two random fish from each group were anesthetized and the liver, head kidney and brain were collected for mRNA isolation and analysis. In the control fish, relative quantification analysis based on the p53 mRNA levels in liver (RQ=1.00) showed higher basal levels of p53 mRNA in the head kidney (RQ=1.69), and lower in the brain (RQ=0.41). In all three tissues sampled, p53 mRNA was affected by treatment with B[a]P. Liver tissue showed the greatest induction (RQ=1.53) from base levels (RQ=1.00), followed by brain (RQ=1.36), and head kidney (RQ=1.23). These results confirm that p53 mRNA is generally present at lower levels in differentiated tissues (liver and brain) than in those tissues with cell lines (head kidney), and demonstrate that p53 is moderately inducible by B[a]P in the whitefish. The approach presented here has the advantage of providing rapid and accurate measures of p53 induction in various tissues of fish responding to PAH contaminant exposure.
Wydawca
-
Rocznik
Tom
09
Numer
2
Opis fizyczny
p.139-143,fig.,ref.
Twórcy
autor
  • University of Warmia and Mazury, Sloneczna 45G, 10-723 Olsztyn, Poland
autor
autor
Bibliografia
  • Abbas T, White D, Hui L, Yoshida K, Foster DA, Bargonetti J 2004) Inhibition of Human p53 Basal Transcription by Down-regulation of Protein Kinase C8. J Biol Chem 279: 9970-9977.
  • Bailey GS, Williams DE, Hendricks JD (1996) Fish models for environmental carcinogenesis: the rainbow trout. En­viron Health Perspect 104: 5-21.
  • Bhaskaran A, May D, Rand-Weaver M, Tyler CR (1999) Fish p53 as a possible biomarker for genotoxins in the aquatic environment. Environ Mol Mutagen 33: 177-184.
  • Braithwaite AW, Royds JA, Jackson P (2005) The p53 story: layers of complexity. Carcinogenesis 26: 1161-1169.
  • Brzuzan P, Jurczyk Ł, Łuczyński MK, Góra M (2005) Rela­tive quantification of CYP1A gene expression in whitefish (Coregonus lavaretus) exposed to benzo[a]pyrene. Envi­ronmental Biotechnology 1: 11-15.
  • Burczynski ME, Penning TM (2000) Genotoxic polycyclic aromatic hydrocarbon orthoquinones generated by aldo-keto reductases induce CYP1A1 via nuclear translo­cation of the aryl hydrocarbon receptor. Cancer Res 60: 908-915.
  • Higuchi R, Fockler C, Dollinger G, Watson R (1993) Kin­etic PCR analysis: real-time monitoring of DNA amplifi­cation reactions. Biotechnology 11: 1026-1030.
  • Hofseth LJ, Hussain SP, Harris CC (2004) p53: 25 years after its discovery. Trends Pharmacol Sci 25: 177-181.
  • Krause MK, Rhodes LD, Van Beneden RJ (1997) Cloning of the p53 tumor suppressor gene from the Japanese medaka (Oryzias latipes) and evaluation of mutational hotspots in MNNG-exposed fish. Gene 189: 101-106.
  • Larkin P, Knoebl I, Denslow ND (2003) Differential gene expression analysis in fish exposed to endocrine disrupt­ing chemicals. Comp Biochem Physiol B136: 149-161.
  • Livak KJ, Schmittgen TD (2001) Analysis of relative gene expression data using Real-Time Quantitative PCR and the 2'aact method. Methods 25: 402-408.
  • Soussi T, de Fromentel C, May P (1990) Structural aspects of the p53 protein in relation to gene evolution. On­cogene 5: 945-952.
Typ dokumentu
Bibliografia
Identyfikatory
Identyfikator YADDA
bwmeta1.element.agro-article-8fe979f7-7c97-401c-af3a-d5470ba07053
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