title: Analysis of point mutation in exon 2 of CYP2E1 gene in renal cell/urothelial cancer patients in comparison with control population.
abstract: OBJECTIVE
  Genetic polymorphisms of human cytochrome P450s have been implicated to be of importance for susceptibility to different cancers. Recently, a point mutation was found in the exon 2 of the CYP2E1 gene (CYP2E1*2) [Hu et al. 1997]. In order to evaluate a possible link between the point mutation in exon 2 of the CYP2E1 gene and the susceptibility to renal cell/urothelial cancer, we developed a screening method based on the polymerase chain reaction (PCR) and restriction fragment length polymorphism (RFLP).
  
  
  METHODS
  DNA of peripheral white blood cells was isolated from 158 renal cell/urothelial cancer patients as well as from 150 controls.
  
  
  METHODS
  Primers for PCR were designed by the Primer 3 release 0.1 program. The PCR yield a product of 215 base pairs (bp), which was digested with the restriction enzyme Hha I. The DNA fragments were separated on a 3% agarose gel stained with ethidium bromide. Restriction enzyme digestion of the PCR product obtained from the wild-type DNA resulted in the appearance of a 66 bp, a 43 bp, a 40 bp, a 39 bp and a 28 bp DNA fragment. In contrast to the wild-type, the digestion of the PCR product from DNA carrying the point mutation resulted in the loss of the 39 bp and 40 bp fragments and the appearance of an additional 79 bp fragment. Therefore, the loss of one Hha I restriction site caused by a single nucleotide exchange is suitable for the identification of the point mutation in exon 2 of CYP2E1 gene.
  
  
  RESULTS
  However, we could not detect any point mutation in any of the 158 renal cell/urothelial cancer patients or the 150 controls. The distribution of the point mutation in exon 2 of CYP2E1 gene did not show any difference in renal cell/urothelial cancer patients and controls.
  
  
  CONCLUSIONS
  This might indicate a lack of association between this CYP2E polymorphism (CYP2E1*2) and renal cell/urothelial cancer.
authors.affiliation: Institute of Clinical Pharmacology, Jena, Germany.
authors.forename: K
authors.initials: K
authors.lastname: Farker
authors.forename: M H
authors.initials: MH
authors.lastname: Lehmann
authors.forename: R
authors.initials: R
authors.lastname: Kästner
authors.forename: J
authors.initials: J
authors.lastname: Weber
authors.forename: V
authors.initials: V
authors.lastname: Janitzky
authors.forename: J
authors.initials: J
authors.lastname: Schubert
authors.forename: A
authors.initials: A
authors.lastname: Hoffmann
mesh_terms: D001483:Base Sequence; D002292:Carcinoma, Renal Cell / Q000097:blood / Q000201:enzymology / Q000235:genetics*; D019392:Cytochrome P-450 CYP2E1 / Q000235:genetics*; D005091:Exons; D020022:Genetic Predisposition to Disease; D005820:Genetic Testing / Q000379:methods; D006801:Humans; D007680:Kidney Neoplasms / Q000097:blood / Q000201:enzymology / Q000235:genetics*; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D012150:Polymorphism, Restriction Fragment Length; D014571:Urologic Neoplasms / Q000097:blood / Q000201:enzymology / Q000235:genetics*
chemical_list: D019392:Cytochrome P-450 CYP2E1