title: Polymorphisms and functions of the aldose reductase gene 5' regulatory region in Chinese patients with type 2 diabetes mellitus.
abstract: OBJECTIVE
  To screen the 5' regulatory region of the aldose reductase (AR) gene for genetic variabilities causing changes in protein expression and affecting the promoter function.
  
  
  METHODS
  The screenings were carried out by polymerase chain reaction-single strand conformation polymorphism (PCR-SSCP). All SSCP variants were submitted for DNA sequencing and inserted into the plasmid chloromycetin acetyl transferase (CAT) enhancer vector. The constructs were used to transfect Hela cells, and CAT assays were performed to assess promoter activity. Gel mobility shift and footprinting assays were also performed to determine the interaction between the DNA and nuclear proteins.
  
  
  RESULTS
  Two polymorphisms, C (-106) T and C (-12) G, were identified in the regulatory region in 123 Chinese control subjects and 145 patients with type 2 diabetes mellitus. The frequencies of genotypes WT/WT, WT/C (-12) G and WT/C (-106) T were not significantly different between the subjects and patients. In the patients with and without retinopathy, frequencies of WT/C (-106) T were 31.5% and 17.5% (P < 0.05) respectively, and the frequencies of WT/C (-12) G were 10.5% and 2.5% (P > 0.05) respectively. The total frequency of WT/C (-12) G and WT/C (-106) T in patients with retinopathy was 41.8%, significantly higher than that (20.0%) in patients without retinopathy (P < 0.025). The relative transcription activities of the wild-type, the C (-12) G and the C (-106) T were 15.7%, 31.0% and 32.2%, respectively. The results of DNA-protein interaction assays showed that these variations did not change the binding site of DNA with trans-acting factors.
  
  
  CONCLUSIONS
  The polymorphisms C (-12) G and C (-106) T strongly associated with diabetic retinopathy in the Chinese population have been identified in the regulatory region of the aldose reductase gene.
authors.affiliation: Department of Biochemistry, Xiangya School of Medicine of Central South University, Changsha 410078, China.
authors.forename: Qingjie
authors.initials: Q
authors.lastname: Li
authors.forename: Ping
authors.initials: P
authors.lastname: Xie
authors.forename: Jianjun
authors.initials: J
authors.lastname: Huang
authors.forename: Yapeng
authors.initials: Y
authors.lastname: Gu
authors.forename: Weimin
authors.initials: W
authors.lastname: Zeng
authors.forename: Huiping
authors.initials: H
authors.lastname: Song
mesh_terms: D024506:5' Flanking Region / Q000235:genetics*; D000328:Adult; D000449:Aldehyde Reductase / Q000235:genetics* / Q000378:metabolism; D001665:Binding Sites / Q000235:genetics; D002681:China; D015500:Chloramphenicol O-Acetyltransferase / Q000235:genetics / Q000378:metabolism; D004247:DNA / Q000737:chemistry / Q000235:genetics; D018983:DNA Footprinting; D003924:Diabetes Mellitus, Type 2 / Q000201:enzymology / Q000235:genetics*; D024202:Electrophoretic Mobility Shift Assay; D005260:Female; D006367:HeLa Cells; D006801:Humans; D008297:Male; D008875:Middle Aged; D009154:Mutation; D016133:Polymerase Chain Reaction; D011110:Polymorphism, Genetic; D018807:Polymorphism, Single-Stranded Conformational; D011993:Recombinant Fusion Proteins / Q000235:genetics / Q000378:metabolism; D012045:Regulatory Sequences, Nucleic Acid / Q000235:genetics*; D017422:Sequence Analysis, DNA; D014158:Transcription, Genetic
chemical_list: D011993:Recombinant Fusion Proteins; D004247:DNA; D000449:Aldehyde Reductase; D015500:Chloramphenicol O-Acetyltransferase