LINC00707靶向miR-30c-5p对ox-LDL诱导的人血管平滑肌细胞增殖、迁移及炎症因子的影响

Effect of LINC00707 on proliferation,migration and inflammatory factors of human vascular smooth muscle cells induced by ox-LDL by targeting miR-30c-5p

目的探讨LINC00707对氧化型低密度脂蛋白(oixidized low-density lipoprotein,ox-LDL)诱导的人血管平滑肌细胞功能障碍的影响及其作用机制。方法采用ox-LDL诱导人血管平滑肌细胞HVSMCs建立动脉粥样硬化细胞模型,si-NC、si-LINC00707、miR-NC、miR-30c-5p mimic分别转染至HVSMCs后,加入100 mg·L-1 ox-LDL处理细胞,si-LINC00707和anti-miR-NC、si-LINC00707和miR-30c-5p Inhibitor分别共转染至HVSMCs后,加入100 mg·L-1 ox-LDL处理细胞;qRT-PCR法检测LINC00707、miR-30c-5p的表达量;CCK-8法、Transwell实验分别检测细胞增殖及迁移;ELISA法检测IL-6、TNF-α、IL-10的水平;双荧光素酶报告实验检测LINC00707与miR-30c-5p的靶向关系;Western blot检测E-cadherin、N-cadherin蛋白表达量。结果ox-LDL诱导的HVSMCs中LINC00707的表达量升高(P<0.05),miR-30c-5p的表达量降低(P<0.05);转染si-LINC00707或miR-30c-5p mimic后,细胞存活率、N-cadherin蛋白水平和IL-6、TNF-α的水平降低(P<0.05),迁移细胞数减少(P<0.05),E-cadherin蛋白水平和IL-10的水平升高(P<0.05);LINC00707可靶向结合miR-30c-5p;共转染si-LINC00707和miR-30c-5p Inhibitor后,细胞存活率、N-cadherin蛋白水平和IL-6、TNF-α的水平升高(P<0.05),迁移细胞数增多(P<0.05),E-cadherin蛋白水平和IL-10的水平降低(P<0.05)。结论下调LINC00707导致miR-30c-5p表达升高从而抑制ox-LDL诱导的HVSMCs增殖、迁移及炎症反应。

Aim To explore the effect of LINC00707 on the proliferation,migration and inflammatory factors of human vascular smooth muscle cells induced by oixidized low-density lipoprotein(ox-LDL)and its possible mechanism.Methods ox-LDL was used to induce human vascular smooth muscle cells(HVSMCs)to establish an atherosclerotic cell model.si-NC,si-LINC00707,miR-NC,miR-30c-5p mimic were transfected into HVSMCs,and then 100 mg·L-1 ox-LDL was added to the cells.si-LINC00707 and anti-miR-NC,or si-LINC00707 and miR-30c-5p Inhibitor were co-transfected into HVSMCs and then treated with 100 mg·L-1 ox-LDL.qRT-PCR was used to detect the expression of LINC00707 and miR-30c-5p.CCK-8 and Transwell test were used to detect cell proliferation and migration.ELISA was used to detect the levels of IL-6,TNF-α,and IL-10.The dual-luciferase reporter experiment was used to detect the targeting relationship between LINC00707 and miR-30c-5p.Western blot was used to detect the protein expression of E-cadherin and N-cadherin.Results The expression of LINC00707 in HVSMCs induced by ox-LDL increased(P<0.05),while the expression of miR-30c-5p decreased(P<0.05).After transfection with si-LINC00707 or miR-30c-5p mimic,cell viability,the protein level of N-cadherin,the levels of IL-6 and TNF-αdecreased(P<0.05),and the number of migrating cells decreased(P<0.05),while the protein level of E-cadherin and the level of IL-10 increased(P<0.05).LINC00707 could target miR-30c-5p.After co-transfection with si-LINC00707 and miR-30c-5p inhibitor,cell survival rate,the protein level of N-cadherin,the levels of IL-6 and TNF-αincreased(P<0.05),and the number of migrating cells increased(P<0.05),while the protein level of E-cadherin and the level of IL-10 decreased(P<0.05).Conclusion Down-regulation of the expression of LINC00707 could inhibit the proliferation,migration and inflammation of human vascular smooth muscle cells induced by ox-LDL by promoting the expression of miR-30c-5p.