circNT5E靶向miR-377-3p对ox-LDL诱导的血管平滑肌细胞增殖、迁移侵袭的影响

Effect of circNT5E targeting miR-377-3p on the proliferation,migration and invasion of vascular smooth muscle cells induced by ox-LDL

目的探讨环状RNA NT5E(circNT5E)对氧化低密度脂蛋白(ox-LDL)诱导的血管平滑肌细胞增殖、迁移侵袭的影响,并探讨其机制是否与调控miR-377-3p表达有关。方法将人主动脉血管平滑肌细胞(HASMCs)分为对照(Con)组(正常培养的细胞)、ox-LDL组(50μg/ml ox-LDL)、ox-LDL+si-NC组(转染小干扰RNA阴性对照,50μg/ml ox-LDL)、ox-LDL+si-circNT5E组(转染sicircNT5E,50μg/ml ox-LDL)、ox-LDL+miR-NC组(转染miR-NC,50μg/ml ox-LDL)、ox-LDL+miR-377-3p组(转染miR-377-3p模拟物,50μg/ml ox-LDL)、ox-LDL+si-circNT5E+anti-miR-NC组(转染si-circNT5E和anti-miR-NC,50μg/ml ox-LDL)、ox-LDL+si-circNT5E+anti-miR-377-3p组(转染sicircNT5E和anti-miR-377-3p,50μg/ml ox-LDL)。实时定量PCR(RT-qPCR)检测circNT5E和miR-377-3p表达;细胞计数法、Transwell实验检测细胞活力、迁移和侵袭细胞数。双荧光素酶报告实验和RTqPCR确定circNT5E对miR-377-3p的靶向作用。结果ox-LDL处理的HASMCs中circNT5E表达量、细胞活力、迁移和侵袭细胞数显著升高(P<0.05),miR-377-3p表达量显著降低(P<0.05)。干扰circNT5E表达或过表达miR-377-3p可减弱ox-LDL对HASMCs增殖、迁移和侵袭的促进作用(P<0.05)。circNT5E直接结合miR-377-3p,干扰circNT5E促进miR-377-3p表达,而过表达circNT5E抑制miR-377-3p表达(P<0.05)。抑制miR-377-3p表达可逆转干扰circNT5E对ox-LDL诱导的HASMCs增殖、迁移和侵袭的抑制作用(P<0.05)。结论干扰circNT5E表达可抑制ox-LDL诱导的血管平滑肌细胞增殖、迁移侵袭,其机制与上调miR-374a-3p表达有关。

Objective To investigate the effect of circular RNA NT5E(circNT5E)on the proliferation,migration and invasion of vascular smooth muscle cells induced by ox-LDL,and to explore whether its mechanism is related to the regulation of miR-377-3p expression.Methods Human aortic vascular smooth muscle cells(HASMCs)were divided into control(Con)group(normally cultured cells),ox-LDL group(50μg/ml ox-LDL),and ox-LDL+si-NC group(transfection Small interfering RNA negative control,50μg/ml ox-LDL),ox-LDL+sicircNT5E group(transfection si-circNT5E,50μg/ml ox-LDL),ox-LDL+miR-NC group(transfection miRNC,50μg/ml ox-LDL),ox-LDL+miR-377-3p group(transfected miR-377-3p mimic,50μg/ml ox-LDL),ox-LDL+si-circNT5E+anti-miR-NC group(transfected with si-circNT5E and anti-miR-NC,50μg/ml ox-LDL),ox-LDL+si-circNT5E+anti-miR-377-3p group(transfected with si-circNT5E and anti-miR-377-3p,50μg/ml ox-LDL).Real-time quantitative PCR(RT-qPCR)was used to detect the expression of circNT5E and miR-377-3p;cell counting method and Transwell experiment were used to detect cell viability,migration and number of invasive cells.The dual luciferase reporter experiment and RT-qPCR confirmed the targeting effect of circNT5E on miR-377-3p.Results The expression of circNT5E,cell viability,migration and invasion cells in HASMCs treated with ox-LDL were significantly increased(P<0.05),and the expression of miR-377-3p was significantly decreased(P<0.05).Interference with circNT5E expression or overexpression of miR-377-3p can attenuate the promotion of ox-LDL on the proliferation,migration and invasion of HASMCs(P<0.05).The circNT5E directly binds to miR-377-3p and interferes with circNT5E to promote the expression of miR-377-3p,while overexpression of circNT5E inhibits the expression of miR-377-3p(P<0.05).Inhibition of miR-377-3p expression can reverse the inhibitory effect of interference with circNT5E on the proliferation,migration and invasion of HASMCs induced by ox-LDL(P<0.05).Conclusion:Interference with the expression of circNT5E can inhibit the proliferation,migration and invasion of vascular smooth muscle cells induced by ox-LDL,and the mechanism is related to the up-regulation of miR-374a-3p expression.