男性miR-221/222基因多态性与心力衰竭的关系

Relationship between MiR-221/222 Gene Polymorphism and Heart Failure in Men

目的:探讨男性miR-221/222基因多态性对其表达的影响,并初步分析该基因与心力衰竭的关系.方法:分别收集男性心力衰竭患者及健康对照外周血,分离血细胞和血浆.提取外周血白细胞基因组DNA,经PCR扩增目的DNA片段、Sanger测序等分别统计相应SNP基因型在各组中的分布差异.分别提取各组血浆miRNA,采用TaqMan探针法检测miR-221/222的表达强度在心力衰竭及对照组中的差异.分别按照miR-221/222基因簇SNP位点进行分组,观察心力衰竭患者中这些变异位点对血浆miR-221/222表达水平的影响.结果:miR-221/222基因簇周围SNP位点rs2858059、rs2858060、rs2858061及rs34678647的各基因型在心衰及对照组中分布无显著差异.rs113054794位点在所有观察对象中仅有C/-基因型.miR-221/222靶基因p27及DDIT4的3'非翻译区miR-221/222结合区SNP位点rs182069485及rs72808106在所有观察对象中均分别为G/G基因型和A/A基因型,无显著多态性.血浆miR-221/222在心衰患者中均升高,且miR-222基因rs34678647位点T/-基因型患者miR-222表达水平较G/-基因型更高.结论:血浆miR-221/222在男性心衰患者中升高,miR-222基因rs34678647的SNP位点与血浆miR-222水平有关.

Objective:To investigate the regulation of miR-221/222 on the expression and the relation with heart failure.Methods:The peripheral blood samples from male heart failure patients and healthy volunteers were collected and separated for blood cells and plasma,respectively.The genome DNA was extracted,amplified and sequenced for analyzing the distribution of SNP genotypes in the heart failure patients and healthy controls.The plasma miRNA was extracted and the expression of miR-221/222 was detected with a TaqMan assay.The plasma levels of miR-221/222 were analyzed according to the SNP genotypes in heart failure.Results:The distribution of SNP genotypes of rs2858059、rs2858060、rs2858061 and rs34678647 were all comparable between heart failure patients and healthy controls.The genotype of rs113054794 was only C/-in all of the study subjects.The rs182069485 and rs72808106 in the 3'UTR of p27 and DDIT4 encoding genes showed no polymorphism in all of the subjects.The plasma levels of miR-221 and miR-222 were increased in heart failure patients.More importantly,we observed that the T/-genotype of rs34678647 in miR-222 was related with the increase of plasma miR-222 in heart failure patients.Conclusion:Plasma miR-221/222 levels are increased in male heart failure patients.The genotype of rs34678647 is related with the plasma miR-222 levels in heart failure patients.