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Relative quantification of Cytochrome P450 1B1 gene expression in peripheral leukocytes using lightcycler
Simone Helmig1, Bahar Hadzaad, Juliane Döhrel
1Institut für Arbeits- und Sozialmedizin der Justus- Liebig Universität, Aulweg 129/III, D-35385 Giessen, Germany. Simone.Helmig@arbmed.med.uni-giessen.de
Background:
The cytochrome P450 oxidase system is a multigene family of inducible enzymes that play a central role in the metabolic activation of various xenobiotics, including polycyclic hydrocarbons (PAH). To investigate the considerable variability of cytochrome P450 1B1 (Cyp1B1) expression due to the exogenous influence of tobacco smoke or the endogenous influence of genetic polymorphism, a sensitive quantitative determination of gene expression is necessary.
Materials And Methods:
A method is introduced for the analysis of Cyp 1B1 gene expression using real-time quantitative PCR and the comparative DeltaDeltaCT (threshold cycle) method. Blood samples from a smoker and non smoker were collected and total RNA was analysed in comparison to the housekeeping gene glyceraldehyde-3-phosphate dehydrogenase (GAPDH).
Results:
The expression of Cyp 1B1 was 16.43 times higher in the smoker than in the non-smoker.
Conclusion:
This approach provides a manageable method for examining large quantities of samples and could possibly be used to evaluate gene environmental interactions on the basics of gene expression analysis.
