title: Association of rare SPINK1 gene mutation with another base substitution in chronic pancreatitis patients.
abstract: OBJECTIVE
  To verify and expand the known spectrum of serine protease inhibitor Kazal type 1 (SPINK1) gene mutations in chronic pancreatitis.
  
  
  METHODS
  DNA extracted from 172 chronic pancreatitis patients was assayed for SPINK1 gene mutations by PCR and DNA sequencing. A control cohort of 90 unrelated healthy individuals was analysed by the same methods for presence of common populational polymorphisms, and frequency of five-loci haplotypes was calculated. Linkages of gene aberrations in single SPINK1 gene copies were analysed by long-distance PCR followed by allele-specific PCR and DNA sequencing.
  
  
  RESULTS
  The most frequent SPINK1 gene mutation N34S was found at a frequency of 6%. Furthermore, we detected the heterozygous intervening sequence (IVS) 3 + 2 T > C mutated gene in 2 German patients and 1 Macedonian chronic pancreatitis patient. In all three SPINK1 gene copies an additional rare base substitution was found: 5'untranslated region (UTR)-215 G > A. Polymorphism analysis revealed that all three affected genes carried the same five-loci haplotype. DNA sequencing of another chronic pancreatitis-related gene PRSS1 (cationic trypsinogen) did not reveal any mutations in these 3 patients.
  
  
  CONCLUSIONS
  We found in 3 (2%) of 172 chronic pancreatitis patients an IVS3 + 2 T > C SPINK1 gene mutation and a base substitution 5'UTR-215 G > A in the same gene copy. Most probably the 5'UTR-215 G > A represents a rare polymorphism and not a mutation as previously concluded. Haplotype analysis suggests a common origin of the IVS3 + 2 T > C mutation in these patients.
authors.affiliation: Department of General, Visceral and Thoracic Surgery, University Medical Center Hamburg-Eppendorf, Martinistrasse 52, Hamburg 20246, Germany.
authors.forename: Viacheslav N
authors.initials: VN
authors.lastname: Kalinin
authors.forename: Jussuf T
authors.initials: JT
authors.lastname: Kaifi
authors.forename: Heidi
authors.initials: H
authors.lastname: Schwarzenbach
authors.forename: Anatoly S
authors.initials: AS
authors.lastname: Sergeyev
authors.forename: Bjoern C
authors.initials: BC
authors.lastname: Link
authors.forename: Dean
authors.initials: D
authors.lastname: Bogoevski
authors.forename: Yogesh
authors.initials: Y
authors.lastname: Vashist
authors.forename: Jakob R
authors.initials: JR
authors.lastname: Izbicki
authors.forename: Emre F
authors.initials: EF
authors.lastname: Yekebas
mesh_terms: D020121:5' Untranslated Regions; D001483:Base Sequence; D002352:Carrier Proteins / Q000235:genetics*; D015331:Cohort Studies; D004247:DNA / Q000378:metabolism; D004252:DNA Mutational Analysis; D005192:Family Health; D005786:Gene Expression Regulation; D020022:Genetic Predisposition to Disease; D006239:Haplotypes; D006801:Humans; D008969:Molecular Sequence Data; D009154:Mutation; D050500:Pancreatitis, Chronic / Q000175:diagnosis* / Q000235:genetics*; D014359:Trypsin Inhibitor, Kazal Pancreatic
chemical_list: D020121:5' Untranslated Regions; D002352:Carrier Proteins; C486642:SPINK1 protein, human; D014359:Trypsin Inhibitor, Kazal Pancreatic; D004247:DNA