2型糖尿病线粒体ND1基因3个新突变位点的分析  

Analysis of three novel mutations of mitochondrial DNA ND1 gene in patients with type 2 diabetes mellitus

作　　者：刘松梅[1] 周新[1] 李霞[1] 秦汉[2] 鲁敏翔[1] 李东[1] 古今刚[1] 

机构地区：[1]武汉大学中南医院检验科 [2]黄石理工学院医学部

出　　处：《中华检验医学杂志》2005年第8期821-824,共4页Chinese Journal of Laboratory Medicine

基　　金：武汉市攻关课题资助项目(20016009107);湖北省自然科学基金资助项目(2004ABA163)

摘　　要：目的探讨线粒体DNAtRNALeu(UUR)基因和ND1基因点突变(np3243、3316、3394和3593)与湖北人群2型糖尿病的相关性。方法采用聚合酶链反应限制性片段长度多态性(PCRRFLP),克隆和测序的分析方法,对184例2型糖尿病患者和210名健康对照进行筛选,并用DNAMAN、Primer、mfold和Antherprot软件对检出的突变位点进行分析和二级结构预测。结果实验组检出3316(G→A)突变6例(3.3%),3394(T→C)突变5例(2.7%),3593(T→C)突变1例(0.5%),并发现3个尚未见报道的新突变位点:3606(A→G)、3618(T→C)和3688(G→C),未检出3243(A→G)突变;对照组只检出3316(G→A)突变1例(0.5%)。两组间仅3394(T→C)突变率差异有统计学意义(P<0.05)。3606(A→G)和3618(T→C)突变分别为亮氨酸、苯丙氨酸的无意义突变,其二级结构均与野生型ND1相同;3316(G→A)突变(丙氨酸→苏氨酸),3394(T→C)突变(酪氨酸→组氨酸),3593(T→C)突变(缬氨酸→丙氨酸),3688(G→C)突变(丙氨酸→脯氨酸),均引起ND1基因DNA和蛋白质二级结构的改变,其中3394(T→C)突变型变化最显著。结论线粒体DNAND1基因3394(T→C)突变以及3688(G→C)突变伴随3316(G→A)突变,可能与2型糖尿病的发生发展有关。Objective To explore the prevalence of various mitochondrial gene mutations （np 3 243,3 316,3 394, 3 593） in patients with type 2 diabetes mellitus（T2DM） in Hubei. Methods 184 cases of T2DM and 210 healthy controls were determined by PCR-restriction fragment length polymorphism and DNA sequencing. All mutations were analyzed by DNAMAN, Primer , mfold and Antherprot softwares. Results In diabetic group, there were 6 carriers （3.3%） of 3 316 G→A （Ala→Thr） mutation,5 （2.7%） of 3 394 T→C （Tyr→His） mutation , 1 （0.5%） of 3 593 T→C（Val→Ala） and none carrier of 3 243 A→G （Glu→Gly） mutation were observed. Meanwhile, three novel mutations of 3 606 A→G（ Leu→Leu） ,3 618 T→C（Phe→Phe） and 3 688 G→C （Ala→Pro）were found. In controls, only 3 316 G→A mutation was found in 1 subject （0. 5% ）. There was significant difference between two groups for np3 394 mutation frequencies （ P 〈 0. 05 ）. The secondary structure prediction revealed that there were differences between np3 316,3 394,3 593,3 688 mutant and wild-type ND1 protein and DNA. Conclusion mtDNA ND1 gene mutations at np 3 394 , 3 316 companied with np 3 688 might implicate in the pathogenesis of T2DM.

关 键 词：DNA 线粒体 突变 糖尿病 非胰岛素依赖型 2型糖尿病患者 线粒体DNA 新突变位点 ND1基因 聚合酶链反应-限制性片段长度多态性 基因3 蛋白质二级结构 

分 类 号：R587.1[医药卫生—内分泌] R743.4[医药卫生—内科学]