title: Transcriptional regulation of the lactase-phlorizin hydrolase gene by polymorphisms associated with adult-type hypolactasia.
abstract: OBJECTIVE
  The mechanism of the developmental downregulation of the lactase-phlorizin hydrolase (LPH) gene underlying adult-type hypolactasia is unknown. We have determined the functional significance of the recently identified two single nucleotide polymorphisms (SNPs), C/T(-13910) and G/A(-22018), associated with adult-type hypolactasia by studying LPH mRNA levels in intestinal biopsy samples with different genotypes.
  
  
  METHODS
  Intestinal biopsy samples were taken from 52 patients with abdominal complaints. Hypolactasia was diagnosed by determining lactase and sucrase activities and calculating their ratio (L/S ratio). The functional effect of the C/T(-13910) and G/A(-22018) genotype on expression of LPH mRNA was demonstrated in patients heterozygous for the C/T(-13910) and G/A(-22018) polymorphism and an informative expressed SNP located in the coding region of the LPH mRNA. Reverse transcription-polymerase chain reaction followed by solid phase minisequencing was used for accessing the relative expression levels of the LPH alleles using informative SNPs located in exons 1, 2, 6, 10, 13, or 17 as markers.
  
  
  RESULTS
  Statistically significant differences between the three different genotypes CC(-13910) GG(-22018), CT(-13910) GA(-22018), and TT(-13910) AA(-22018) and their respective L/S ratios were observed. Relative quantitation of the expressed LPH alleles showed that the persistent allele represented 92 (6)% (mean (SEM), range 78-99%; n=14) of the expressed LPH mRNA. The patient with the homozygous persistent TT(-13910) AA(-22018), as well as hypolactasic patients with CC(-13910) GG(-22018), showed equal expression of both alleles (47 (1)%; n=7).
  
  
  CONCLUSIONS
  Expression of LPH mRNA in the intestinal mucosa in individuals with T(-13910) A(-22018) alleles is several times higher than that found in individuals with C(-13910), G(-22018) alleles. These findings suggest that the two SNPs, C/T(-13910) and G/A(-22018), associated with adult-type hypolactasia, are associated with the transcriptional regulation of the LPH gene. The presence of the T(-13910) A(-22018) allele also shows significant elevation of the L/S ratio.
authors.affiliation: Department of Medical Genetics, University of Helsinki, Finland.
authors.forename: M
authors.initials: M
authors.lastname: Kuokkanen
authors.forename: N S
authors.initials: NS
authors.lastname: Enattah
authors.forename: A
authors.initials: A
authors.lastname: Oksanen
authors.forename: E
authors.initials: E
authors.lastname: Savilahti
authors.forename: A
authors.initials: A
authors.lastname: Orpana
authors.forename: I
authors.initials: I
authors.lastname: Järvelä
mesh_terms: D000328:Adult; D000483:Alleles; D015971:Gene Expression Regulation, Enzymologic / Q000235:genetics; D005838:Genotype; D006801:Humans; D007413:Intestinal Mucosa; D043322:Lactase; D010694:Lactase-Phlorizin Hydrolase / Q000097:blood / Q000235:genetics*; D020641:Polymorphism, Single Nucleotide / Q000235:genetics*; D012333:RNA, Messenger / Q000032:analysis; D020133:Reverse Transcriptase Polymerase Chain Reaction / Q000379:methods; D013393:Sucrase / Q000097:blood; D014158:Transcription, Genetic / Q000235:genetics; D001616:beta-Galactosidase / Q000097:blood*
chemical_list: D012333:RNA, Messenger; D043322:Lactase; D001616:beta-Galactosidase; D013393:Sucrase; D010694:Lactase-Phlorizin Hydrolase