Polymorphisms and functions of the aldose reductase gene 5′ regulatory region in Chinese patients with type 2 diabetes mellitus

Qingjie Li, Ping Xie, Jianjun Huang, Yapeng Gu, Weimin Zeng, Huiping Song

Research output: Contribution to journalArticle

37 Citations (Scopus)

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. Conclusion. 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.

Original languageEnglish (US)
Pages (from-to)209-213
Number of pages5
JournalChinese Medical Journal
Volume115
Issue number2
StatePublished - 2002
Externally publishedYes

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Aldehyde Reductase
Nucleic Acid Regulatory Sequences
Type 2 Diabetes Mellitus
Genes
Chloramphenicol
Transferases
DNA
Single-Stranded Conformational Polymorphism
Trans-Activators
Electrophoretic Mobility Shift Assay
Diabetic Retinopathy
Nuclear Proteins
DNA Sequence Analysis
HeLa Cells
Proteins
Plasmids
Gels
Binding Sites
Genotype
Polymerase Chain Reaction

Keywords

  • Aldose reductase
  • CAT reporter assay
  • Gene
  • Polymorphism
  • Retinopathy
  • Type 2 diabetes mellitus

ASJC Scopus subject areas

  • Medicine(all)

Cite this

Polymorphisms and functions of the aldose reductase gene 5′ regulatory region in Chinese patients with type 2 diabetes mellitus. / Li, Qingjie; Xie, Ping; Huang, Jianjun; Gu, Yapeng; Zeng, Weimin; Song, Huiping.

In: Chinese Medical Journal, Vol. 115, No. 2, 2002, p. 209-213.

Research output: Contribution to journalArticle

Li, Qingjie ; Xie, Ping ; Huang, Jianjun ; Gu, Yapeng ; Zeng, Weimin ; Song, Huiping. / Polymorphisms and functions of the aldose reductase gene 5′ regulatory region in Chinese patients with type 2 diabetes mellitus. In: Chinese Medical Journal. 2002 ; Vol. 115, No. 2. pp. 209-213.
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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. Conclusion. 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.",
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TY - JOUR

T1 - Polymorphisms and functions of the aldose reductase gene 5′ regulatory region in Chinese patients with type 2 diabetes mellitus

AU - Li, Qingjie

AU - Xie, Ping

AU - Huang, Jianjun

AU - Gu, Yapeng

AU - Zeng, Weimin

AU - Song, Huiping

PY - 2002

Y1 - 2002

N2 - 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. Conclusion. 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.

AB - 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. Conclusion. 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.

KW - Aldose reductase

KW - CAT reporter assay

KW - Gene

KW - Polymorphism

KW - Retinopathy

KW - Type 2 diabetes mellitus

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M3 - Article

VL - 115

SP - 209

EP - 213

JO - Chinese Medical Journal

JF - Chinese Medical Journal

SN - 0366-6999

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