Long non-coding RNA LOC285194 (LOC285194) has reported to regulate vascular smooth muscle cells (VSMCs) proliferation and apoptosis in vitro and in vivo. 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.

中文翻译：

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长非编码 RNA LOC285194 通过靶向 miR-211/PUMA 和 TGF-β1/S100A4 信号抑制血管平滑肌细胞的增殖和迁移，但促进细胞凋亡。

据报道，长非编码 RNA LOC285194 (LOC285194) 可在体外和体内调节血管平滑肌细胞 (VSMC) 增殖和凋亡。本研究旨在探讨LOC285194在VSMC增殖、迁移和凋亡中的作用及其潜在机制。A7r5细胞用Lv-LOC285194或对照Lv-NC转染24-72小时，或靶向S100A4的小干扰RNA(S100A4 siRNA)转染24-48小时，或用Lv-LOC285194和PUMA siRNA共转染72小时，或用miR-211抑制剂处理或用Lv-LOC285194和miR-211模拟物共转染72小时。Lv-LOC285194 转染后，A7r5 细胞还用转化生长因子 - β(TGF-β) (5 ng/ml) 处理 24 小时。使用荧光素酶报告基因测定证实了 LOC285194 和 TGF-β 之间的关系。通过细胞计数试剂盒-8 (CCK-8) 测定、ELISA 和 TUNEL 染色分析细胞增殖和细胞凋亡。通过 qPCR 检测检测 LOC285194 和 miR-211 表达。Western blot法检测S100A4、促凋亡蛋白和抗凋亡蛋白。LOC285194 抑制细胞增殖、侵袭和迁移，促进细胞凋亡，并伴有 PUMA 上调和 miR-211 和 S100A4 下调。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上调并减少LOC285194诱导的细胞凋亡。TGF-β (5 ng/ml) 治疗可逆转 S100A4 siRNA 或 LOC285194 诱导的 S100A4 表达。荧光素酶报告基因检测显示 TGF-β 是 LOC285194 的靶标。LOC285194通过靶向miR-211/PUMA信号抑制血管平滑肌细胞增殖并促进细胞凋亡；此外，LOC285194 通过靶向 TGF-β1/S100A4 信号减少细胞侵袭和迁移。