摘要
目的观察微小RNA-218（miR-218）对大鼠主动脉平滑肌细胞（VSMCs）表型转化、增殖及迁移的作用，探讨miR-218影响VSMCs生物学行为的机制。方法20％胎牛血清（FBS）培养大鼠VSMCs，将miR-218模拟物、miR-218抑制剂及阴性对照转染VSMCs。检测miR-218和表型蛋白α-肌动蛋白（α-actin）、调宁蛋白1（Calponin-1）的含量变化。10％FBS培养条件下，检测miR-218组和miR-nc组的细胞增殖和迁移能力变化，并与miR-218抑制剂组比较。结果20％FBS培养VSMCs3d和7d，miR-218表达量均下调超过90％（P〈0．01），α-actin和Calponin-1表达亦显著下调；与miR-nc组比较，miR-218组α-actin和Calponin-1的表达无明显下调。10％FBS培养VSMCs2-3d，miR-218组VSMCs的增殖力和迁移力均下调28％（P〈0．05）；而miR-218抑制剂组VSMCs的增殖力上调20％（P〈0．05）、迁移力上调53％（P〈0．01）。10％FBS培养VSMCs48～96h，miR-218组中miR-218潜在靶基因周期蛋白依赖性激酶6（CDK6）的表达明显下调。结论miR-218表达上调可抑制大鼠VSMCs的表型转化、增殖和迁移，其作用可能是通过靶向CDK6来实现的。
Objective To observe microRNA （miR）-218 regulation in phenotype transformation, proliferation and migration of rat aortic vascular smooth muscle cells （VSMCs） and explore the mechanism of miR-218 affecting the biological behaviors of VSMCs. Methods Rat aortic VSMCs were cultured in 20% fatal bovine serun（FBS）, and transfected with miR-218 mimics, miR-218 inhibitor and miR-nc for negative control. The expression of miR-218 and α-actin and Calponin-1 in VSMCs was detected. In VSMCs cultured in 10% FBS, VSMCs proliferation and migration in both miR-218 mimics and miR-nc groups were evaluated, and results were compared to miR-218 inhibitor group. Results In VSMCs cultured in 20% FBS for 3 days and 7 days, the expression levels of miR-218 were down-regulated more than 90% （P 〈 0. 01 ） , and those of α-actin and Calponin-1 were down-regulated significantly simultaneously. There was no significant down-regulation of α-actin and Calponin-1 in miR-218 mimics group as compared with miR-nc group. In VSMCs cultured in by 10% FBS for 2-3 days, cell proliferation and migration were significantly inhibited by 28% in miR-218 mimics group （P 〈 0. 05 ）, and they were significantly enhanced by 20% （P 〈 0. 05 ） and 53 % （P 〈 0. 01 ） in miR-218 inhibitor group respectively. In VSMCs cultured in 10% FBS 2-4 days, the expression of cyclin dependent kinase 6 （CDK6） , the potential target gene of miR-218, was significantly down-regulated in miR-218 mimics group. Conclusion Our results suggest that up-regulation of miR-218 inhibits phenotype transformation, proliferation and migration of rat VSMCs, which may be achieved partly through targeting the CDK6 expression.
出处
《中华实验外科杂志》
CAS
CSCD
北大核心
2013年第5期901-904,共4页
Chinese Journal of Experimental Surgery
