标题：2型糖尿病患者KCNJ11启动子区甲基化与胰岛β细胞功能的相关性研究

      作者：李雯翀 1，朱咏瑶 1，刘抐岢 1，陈毅光 1，林志坚 2    (东莞市第三人民医院内分泌科 1、检验科 2，广东 东莞 523320)

      卷次： 2017年28卷20期

      【摘要】 目的 探讨 2型糖尿病(T2DM)患者 KCNJ11启动子区甲基化与基因转录表达、胰岛素抵抗指数(Homa-IR)、胰岛β细胞功能指数(Homa-β)等指标的关系。方法 选取2014年4月至2015年4月在东莞市第三人民医院内分泌科初治的T2DM患者 15例，根据糖化血红蛋白(HbA1c)水平，将HbA1c≤9%的患者纳入A组(n=5)，HbA1c>9%者纳入B组(n=10)；选择15例健康者纳入对照组。用亚硫酸盐法测定KCNJ11基因启动子5'端300 bp中的 46个CpG位点DNA甲基化水平，用半定量逆转录聚合酶链反应(RT-PCR)测定 KCNJ11基因转录表达，并对KCNJ11启动子区甲基化与Homa-IR、Homa-β的关系进行相关性分析。结果 对照组受检者的CpG+62、CpG+66、CpG+351甲基化比率分别为(58±24)%、(46±17)%、(96±8)%，A组分别为(60±19)%、(42±18)%、(99±7)%，B组分别为（60±18）%、(41±20)%、(98±4)%，三组比较差异无统计学意义(P>0.05)；T2DM组与对照组mRNA表达分别为（1.27±0.56）、(0.95±0.49)，差异无统计学意义(P>0.05)；B组患者的CpG+66与Homa-IR呈负相关(r=-0.696，P<0.05)，CpG+62与CpG+66甲基化总和则与Homa-β呈负相关(r=-0.664，P<0.05)。结论 2型糖尿病患者KCNJ11启动子区甲基化可能与胰岛β细胞分泌胰岛素的功能相关，并参与2型糖尿病的发病。
      【关键词】 2型糖尿病；DNA甲基化；聚合酶联反应；基因
      【中图分类号】 R587.1 【文献标识码】 A 【文章编号】 1003—6350（2017）20—3287—04

Relationship between the DNA methylation of KCNJ11's promoter and islet β-cell function in patients with type2 diabetes.
LI Wen-chong 1, ZHU Yong-yao 1, LIU Rui-ke 1, CHEN Yi-guang 1, LIN Zhi-jian 2. Department ofEndocrinology 1, Department of Clinical Laboratory 2, The Third People's Hospital of Dongguan, Dongguan 523320,Guangdong, CHINA
【Abstract】 Objective To investigate the relationship between KCNJ11 promoter methylation and gene tran-scription, Homa-IR and Homa-β in patients with type 2 diabetes mellitus (T2DM). Methods A total of 15 new casesof T2DM patients, who admitted Department of Endocrinology of the Third People's Hospital of Dongguan from April2014 to April 2015, were selected and divided into A group (5 cases of HbA1c≤9%) and B group (10 cases of HbA1c>9%) according to the level of HbA1c, and 15 healthy subjects were included into the control group. DNA methylationlevel of 5' end of 300 bp in the 46 CpG loci of KCNJ11 gene promoter was determined by sulfite method, The tran-scriptional expression of KCNJ11 gene was measured by semi-quantitative reverse transcription polymerase chain re-action (RT-PCR). The relationship between KCNJ11 promoter methylation and Homa-IR, Homa-β was analyzed. Re-sults The methylation ratio of CpG+62, CpG+66, CpG+351 in the control group were (58±24)%, (46±17)%, (96±8)%, respectively, (60±19)%, (42±18)%, (99±7)% in A group, and (60±18)%, (41±20)%, (98±4)% in B group, respective-ly, with no significant difference between the three groups (P>0.05). There was no significant difference in the expres-sion of mRNA between T2DM patients (1.27±0.56) and the control group (0.95±0.49) (P>0.05), but in the B group,CpG+66 was negatively correlated with Homa-IR (r=-0.696, P<0.05), and CpG+62 and CpG+66 methylation sum wasnegatively correlated with Homa-β (r=-0.664, P<0.05). Conclusion The methylation of KCNJ11 promoter region intype 2 diabetic patients may be related to the insulin secretion function of islet beta cells, and involved in the morbidityof type 2 diabetes mellitus.
      【Key words】 Type 2 diabetes mellitus (T2DM); DNA methylation; Polymerase chain reaction (PCR); Gene·论 著·doi:10.3969/j.issn.1003-6350.2017.20.007基金项目：东莞市科技计划项目（编号：201410515000819）