Qingdao University, Qingdao, Shandong 266073, P.R. China.
Department of Cardiology, Qingdao Municipal Hospital (Group), Qingdao, Shandong 266034, P.R. China.
Mol Med Rep. 2019 Aug;20(2):1781-1789. doi: 10.3892/mmr.2019.10410. Epub 2019 Jun 21.
GC binding factor 2 (GCF2) is a transcriptional repressor that inhibits the transcription of GC‑rich promoters, thereby regulating biological processes, including proliferation. However, the role of GCF2 in vascular smooth muscle cells (VSMCs) remains unclear. The level of α‑smooth muscle (α‑SM) actin was determined by immunofluorescence. Cell viability, migration and invasion were analyzed using Cell Counting Kit‑8, wound healing and Transwell assays, respectively. Apoptosis and cell cycle progression were determined using flow cytometry. The expressions of Bcl‑2, Bax, cleaved caspase‑3, cyclin E, CDK2 and the CDK inhibitor p21 were determined by reverse transcription‑quantitative (RT‑q)PCR and western blot analysis. RT‑qPCR was performed to analyze the levels of GCF2 and western blot analysis was conducted to determine the phosphorylation levels of PI3K and AKT. α‑SM actin was found to be expressed in VSMCs. Cell viability, migration and invasion were inhibited by small interfering (si)RNA targeting GCF2. Changes in the expression levels of Bcl‑2, Bax and cleaved caspase‑3 showed that the pro‑apoptotic capacity of the cells was increased by siGCF2. Cell cycle arrest in the G0/G1 phase was induced by siGCF2, which was accompanied by changes in the levels of cyclin E, CDK2 and p21. Furthermore, phosphorylation of PI3K and AKT was suppressed by siGCF2. However, the inhibitory effects of siGCF2 on cell viability, migration and invasion were increased by insulin‑like growth factor 1, which is a specific agonist of AKT. The anti‑proliferative activity of siGCF2 may be associated with the PI3K/AKT pathway in VSMCs.
GC 结合因子 2 (GCF2) 是一种转录抑制因子,可抑制富含 GC 的启动子的转录,从而调节包括增殖在内的生物学过程。然而,GCF2 在血管平滑肌细胞 (VSMC) 中的作用尚不清楚。通过免疫荧光法测定α-平滑肌 (α-SM) 肌动蛋白的水平。使用细胞计数试剂盒-8、划痕愈合和 Transwell 测定分别分析细胞活力、迁移和侵袭。通过流式细胞术测定细胞凋亡和细胞周期进程。通过逆转录-定量 (RT-q)PCR 和 Western blot 分析测定 Bcl-2、Bax、cleaved caspase-3、细胞周期蛋白 E、CDK2 和 CDK 抑制剂 p21 的表达。通过 RT-qPCR 分析 GCF2 的水平,通过 Western blot 分析测定 PI3K 和 AKT 的磷酸化水平。发现 α-SM 肌动蛋白在 VSMC 中表达。靶向 GCF2 的小干扰 (si)RNA 抑制细胞活力、迁移和侵袭。siGCF2 改变 Bcl-2、Bax 和 cleaved caspase-3 的表达水平表明细胞的促凋亡能力增强。siGCF2 诱导细胞周期停滞在 G0/G1 期,同时 cyclin E、CDK2 和 p21 的水平发生变化。此外,siGCF2 抑制 PI3K 和 AKT 的磷酸化。然而,胰岛素样生长因子 1(AKT 的特异性激动剂)增加了 siGCF2 对细胞活力、迁移和侵袭的抑制作用。siGCF2 的抗增殖活性可能与 VSMC 中的 PI3K/AKT 途径有关。