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

作     者：李清解 谢平 黄建军 谷亚鹏 曾卫民 宋惠萍 LI Qingjie;XIE Ping;Huang Jianjun;GU Yapeng;ZENG Weimin;SONG Huiping

作者机构：中南大学湘雅医学院生物化学教研室，长沙，410078 

出 版 物：《Chinese Medical Journal》 (中华医学杂志（英文版）)

年 卷 期：2002年第115卷第2期

页      面：209-213页

核心收录：

学科分类：1002[医学-临床医学] 100201[医学-内科学(含：心血管病、血液病、呼吸系病、消化系病、内分泌与代谢病、肾病、风湿病、传染病)] 10[医学] 

基　　金：国家自然科学基金 

主　　题：5' Flanking Region Adult Aldehyde Reductase Binding Sites China Chloramphenicol O-Acetyltransferase DNA DNA Footprinting Diabetes Mellitus, Type 2 Electrophoretic Mobility Shift Assay Female Hela Cells Humans Male Middle Aged Mutation Polymerase Chain Reaction Polymorphism, Genetic Polymorphism, Single-Stranded Conformational Recombinant Fusion Proteins Regulatory Sequences, Nucleic Acid Research Support, Non-U.S. Gov't Sequence Analysis, DNA Transcription, Genetic 

摘      要：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. 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