微小RNA-22调控人动脉平滑肌细胞增殖、迁移和凋亡功能及其机制  

The roles of microRNA -22 in regulating human artery smooth muscle cells function and mecha-nisms

作　　者：黄水传[1] 许托 黄显莹[1] 刘泽鑫 梁忠锃 张智 Huang Shuichuan;Xu Tuo;Huang Xianying;Liu Zexin;Liang Zhongzeng;Zhang Zhi(Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, Guangzhou 510515, China;Department of Vascular Surgery, Affiliated Hospital of Guang- dong Medical University, Zhanjiang 524001, China)

机构地区：[1]南方医科大学南方医院血管外科,广州510515 [2]广东医科大学附属医院血管外科,湛江524001

出　　处：《中华实验外科杂志》2018年第6期1034-1036,共3页Chinese Journal of Experimental Surgery

基　　金：广东省科技计划项目（2017A020215124）;湛江市科技计划项目（2017801120）

摘　　要：目的探讨微小RNA（miRNA，miR）-22是否参与调控人动脉平滑肌细胞（ASMCs）功能及其机制。方法实时荧光定量聚合酶链反应（FQ—-PCR）和荧光原位杂交观察miR-22在动脉壁中的表达特点；细胞计数试剂盒（CCK-8）法、迁移小室（Transwell）及流式细胞实验分别检测miR-22对ASMCs增殖、迁移及凋亡的影响；双荧光素酶基因报告实验确定miR-22的靶基因；通过慢病毒过表达miR-22后，观察其对大鼠颈动脉球囊损伤后新生内膜的影响。结果（1）miR-22在动脉硬化闭塞症（ASO）血管壁中表达（0．256±O．046）明显低于正常血管壁（0．946±0．081，P=0．000），且其主要定位于动脉中层；（2）与对照组比较，miR-22具有抑制ASMCs增殖（1．146±0．050比1．677±0．082，P=0．000）和迁移（16．50±1．38比37．83±2．54，P=0．000），促进凋亡的功能[（7．33±0．98）％比（3．95±0．89）％，P=0．000]。（3）双荧光素酶基因报告实验证实真核生物延伸因子2激酶（eEF2K）是miR-22的靶基因，miR-22可在转录后水平负性调控eEF2K表达；（4）慢病毒（LV）-miR-22组（0．832±0．126）与对照组（1．579±0．215，P=0．000）比较，明显减少大鼠颈动脉新生内膜面积。结论miR-22通过靶向结合eEF2K，抑制ASMCs增殖及迁移、促进凋亡，减少新生内膜增生。Objective To explore the roles of microRNA （miRNA, miR） -22 in regulating human artery smooth muscle ceils （ASMCs） function and the mechanisms, Methods Real - time fluorescent quantitative polymerase chain reaction （ FQ - PCR） and fluorescence in situ hybridization were performed to detect miR -22 expression in human arteries. Cell counting kit -8 （CCK -8）, transwell and flow cytometry assays were used to assess the effect of miR - 22 on proliferation, migration and apoptosis of ASMCs, respectively. The luciferase reporter assays were used to verify the target genes of miR - 22. A rat carotid artery balloon - injury model was used to detect the role of miR - 22 in neointima formation. Results The miR -22 expression was significantly downregulated in arteriosclerosis obliterans （ASO） arteries as compared with normal arteries （0. 256±0. 046 vs. 0. 946±0. 081, P =0. 000）. The miR -22 mimic had anti - proliferative （ 1. 146 ±0. 050 vs. 1. 677±0. 082, P = 0. 000）, anti - migratory （16.50±1.38 vs. 37. 83 ±2.54, P=0.000） and pro - apoptotic effects [（7.33±0.98）% vs. （3.95 ±0. 89） %, P =0. 000 ] in ASMCs. The dual - luciferase assay showed that eukaryotic elongation factor 2 kinase （eEF2K） was a direct target of miR -22 in ASMCs and that eEF2K expression was signifi- cantiy downregulated by miR - 22 at a post transcriptional level. The lentiviral - miR -22 - mediated miR - 22 upregulation significantly inhibited neointimal formation （ 0. 832±0. 126 vs. 1. 579±0. 215, P = 0. 000）. Conclusion miR - 22 is an important molecule in regulating ASMCs proliferation, migration and aooDtosis bv tarzeting eEF2K.

关 键 词：血管平滑肌细胞 再狭窄 微小RNA-22 真核生物延伸因子2激酶 

分 类 号：R543.5[医药卫生—心血管疾病]