Department of Emergency Medicine, The Affiliated Hospital of Qingdao University , Qingdao, Shandong, China.
Ultrasound, The Affiliated Hospital of Qingdao University , Qingdao, Shandong, China.
Bioengineered. 2020 Dec;11(1):718-728. doi: 10.1080/21655979.2020.1788354.
Long non-coding RNA LOC285194 (LOC285194) has reported to regulate vascular smooth muscle cells (VSMCs) proliferation and apoptosis and . Here we aimed to determine the role of LOC285194 in the proliferation, migration and apoptosis of VSMCs and its underlying mechanisms. A7r5 cells were transfected with Lv-LOC285194 or control Lv-NC for 24-72 h, or small interfering RNA targeting S100A4 (S100A4 siRNA) for 24-48 h, or co-transfected with Lv-LOC285194 and PUMA siRNA for 72 h, or treated with miR-211 inhibitor or co-transfected with Lv-LOC285194 and miR-211 mimics for 72 h. A7r5 cells were also treated with transforming growth factor - β(TGF-β) (5 ng/ml) after Lv-LOC285194 transfection for 24 h. The relationship between LOC285194 and TGF-β was confirmed using luciferase reporter assay. Cell proliferation and cell apoptosis were analyzed by Cell Counting Kit-8 (CCK-8) assay, ELISA and TUNEL staining. LOC285194 and miR-211 expression were detected by qPCR assay. S100A4, pro-apoptotic and anti-apoptotic protein were detected by Western blot assay. LOC285194 inhibited cell proliferation, invasion and migration and promoted cell apoptosis accompanied by upregulation of PUMA and downregulation of miR-211 and S100A4. Targeting PUMA reversed the effect of LOC285194 on cell apoptosis and proliferation. miR-211 mimic inhibited LOC285194-induced PUMA upregulation and decreased LOC285194-induced cell apoptosis. TGF-β (5 ng/ml) treatment reversed S100A4 siRNA or LOC285194-induced S100A4 expression. Luciferase reporter assay showed that TGF-β was the target of LOC285194. LOC285194 inhibits proliferation and promoted apoptosis in vascular smooth muscle cells via targeting miR-211/PUMA signal; In addition, LOC285194 decreased cell invasion and migration by targeting TGF-β1/S100A4 signal.
长链非编码 RNA LOC285194（LOC285194）已被报道可调节血管平滑肌细胞（VSMCs）的增殖和凋亡。本研究旨在确定 LOC285194 在 VSMCs 增殖、迁移和凋亡中的作用及其潜在机制。用 Lv-LOC285194 或对照 Lv-NC 转染 A7r5 细胞 24-72 小时，或用靶向 S100A4 的小干扰 RNA（S100A4 siRNA）转染 24-48 小时，或共转染 Lv-LOC285194 和 PUMA siRNA 72 小时，或用 miR-211 抑制剂处理或共转染 Lv-LOC285194 和 miR-211 模拟物 72 小时。转染 Lv-LOC285194 24 小时后，A7r5 细胞也用转化生长因子-β（TGF-β）（5ng/ml）处理。用荧光素酶报告基因测定法证实 LOC285194 与 TGF-β 的关系。用细胞计数试剂盒-8（CCK-8）测定、ELISA 和 TUNEL 染色分析细胞增殖和细胞凋亡。用 qPCR 测定 LOC285194 和 miR-211 的表达。用 Western blot 测定 S100A4、促凋亡和抗凋亡蛋白的表达。LOC285194 抑制细胞增殖、侵袭和迁移，促进细胞凋亡，同时上调 PUMA，下调 miR-211 和 S100A4。靶向 PUMA 逆转了 LOC285194 对细胞凋亡和增殖的影响。miR-211 模拟物抑制 LOC285194 诱导的 PUMA 上调，并降低 LOC285194 诱导的细胞凋亡。TGF-β（5ng/ml）处理逆转了 S100A4 siRNA 或 LOC285194 诱导的 S100A4 表达。荧光素酶报告基因测定显示，TGF-β 是 LOC285194 的靶点。LOC285194 通过靶向 miR-211/PUMA 信号抑制血管平滑肌细胞增殖并促进凋亡；此外，LOC285194 通过靶向 TGF-β1/S100A4 信号减少细胞侵袭和迁移。
