Zheng Lu, Zhang Jun, Mu Qinfeng, Zhang Xiaoying, Luo Guanghua
Comprehensive Laboratory, First People's Hospital of Changzhou, Changzhou 213003, China.
Comprehensive Laboratory, First People's Hospital of Changzhou, Changzhou 213003, China. Email:
Zhonghua Xin Xue Guan Bing Za Zhi. 2014 Apr;42(4):284-9.
To investigate the association between genetic polymorphisms of proximal promoter region of apolipoprotein M (apoM) gene and susceptibility of coronary artery diseases (CAD) in Han Chinese population.
Two pairs of primers were designed according to the sequence (GenBank accession nos. EU030444.1) and the PCR products of apoM proximal promoter region were directly sequenced. Two hundred and six patients [165 males, mean age (61.9 ± 9.2) years old] diagnosed with CAD according to the results of angiography (a lesion was classed as being significant when stenosis was more than 50%) were enrolled in the present study, 209 age- and gender-matched patients[157 males, mean age (60.4 ± 9.1) years old] without CAD according to the results of angiography were selected as the control group. The allelic frequencies and genotype distributions of polymorphism in CAD and non-CAD patients were analyzed. Furthermore the wide-type and mutant promoter region of apoM were cloned into the luciferase expression vector pGL3, respectively. Luciferase reporter assay was used to detect the activity of apoM promoter.
A new deletion mutation -724delC in apoM promoter was found. The frequency of Del C allele was 8.0% in CAD patients and only 4.1% in the non-CAD controls (OR = 2.054, 95%CI 1.125-3.749, P = 0.017). The mean TC level was lower in groups with wide-type homozygotes compared to the mutant allele carriers [ (6.04 ± 0.90) mmol/L vs. (4.95 ± 1.00)mmol/L, P < 0.01]. -724delC mutant showed obvious decreased luciferase activities (1.13 ± 0.25 vs. 2.11 ± 0.15, P = 0.009).
It is reasonable to speculate that -724delC could affect the activity of the apoM promoter and downregulate apoM expressions, therefore, influence the susceptibility of CAD in this patient cohort.
探讨载脂蛋白M（apoM）基因启动子近端区域的基因多态性与中国汉族人群冠状动脉疾病（CAD）易感性之间的关联。
根据序列（GenBank登录号：EU030444.1）设计两对引物，对apoM启动子近端区域的PCR产物进行直接测序。本研究纳入了206例根据血管造影结果诊断为CAD的患者[165例男性，平均年龄（61.9±9.2）岁]（当狭窄超过50%时，病变被归类为显著病变），根据血管造影结果选择209例年龄和性别匹配的无CAD患者[157例男性，平均年龄（60.4±9.1）岁]作为对照组。分析CAD患者和非CAD患者中多态性的等位基因频率和基因型分布。此外，将apoM的野生型和突变型启动子区域分别克隆到荧光素酶表达载体pGL3中。采用荧光素酶报告基因检测法检测apoM启动子的活性。
在apoM启动子中发现了一个新的缺失突变-724delC。Del C等位基因在CAD患者中的频率为8.0%，在非CAD对照组中仅为4.1%（OR = 2.054，95%CI 1.125 - 3.749，P = 0.017）。与突变等位基因携带者相比，野生型纯合子组的平均总胆固醇（TC）水平较低[（6.04±0.90）mmol/L对（4.95±1.00）mmol/L，P < 0.01]。-724delC突变体的荧光素酶活性明显降低（1.13±0.25对2.11±0.15，P = 0.009）。
有理由推测-724delC可能影响apoM启动子的活性并下调apoM的表达，因此，影响该患者队列中CAD的易感性。
