MiR-17/RB通路调节血管平滑肌细胞增殖

MiR-17/RB pathway regulates vascular smooth muscle cells proliferation

目的探索miR-17在血管平滑肌细胞(VSMCs)中对视网膜母细胞瘤(RB)基因和蛋白水平的调控作用,以及对增殖效应的影响。方法培养HA-VSMCs并诱导其增殖,应用芯片技术筛选出差异表达水平最显著的微小RNA(miR)。在293T细胞应用荧光素酶报告基因验证生物信息学软件预测的miR-17与RB的靶向结合作用。分别向VSMC转染miR-17 mimic和miR-17 inhibitor并建立阴性对照,检测RB水平及细胞增殖相关指标PCNA表达水平。结果荧光素酶报告分析证实miR-17与RB mRNA-3’UTR具有靶向结合效应。转染miR-17 mimic促进VSMC增殖相关指标PCNA表达上调,并且可以下调RB蛋白及mRNA水平,转染miR-17 inhibitor后下调PCNA表达且可上调RB表达(P<0.05)。结论miR-17通过靶向结合RB mRNA-3’UTR调控RB表达从而调控VSMCs增殖。

Objective This study aimed to explore effects of miR-17 onretinoblastoma(RB) and cell proliferation in vascular smooth muscle cells(VSMCs). Methods Screen differential expressing micro RNA(miRNAs) in induced proliferating HA-VSMCs. Select the most differently expressing miRNA for furthur study. Use luciferase report gene to confirmtargeting effect between miR-17 and RB-3 'UTR. Transfect miR-17 mimic or miR-17 inhibitor to HA-VSMCs and RB level and PCNA level was detected. Results Luciferase report gene results showed miR-17 could target RB mRNA-3'UTR. After transfecting miR-17 mimic to HA-VSMCs, PCNA was upregulated while RB level was downregulated(P<0.05). PCNA was downregulated and RB level was upregulated after transfecting with miR-17 inhibitor(P<0.05). Conclusion miR-17 can regulate RB level and HA-VSMCs proliferation through targeting RB mRNA-3'UTR.