Knockdown of GC binding factor 2 by RNA interference inhibits invasion and migration of vascular smooth muscle cells

Ma,Ying; Ren,Yongqiang; Guan,Jun

doi:10.3892/mmr.2019.10410

Knockdown of GC binding factor 2 by RNA interference inhibits invasion and migration of vascular smooth muscle cells

Authors:
- Ying Ma
- Yongqiang Ren
- Jun Guan
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Affiliations: Qingdao University, Qingdao, Shandong 266073, P.R. China, Department of Cardiology, Qingdao Municipal Hospital (Group), Qingdao, Shandong 266034, P.R. China
Published online on: June 21, 2019 https://doi.org/10.3892/mmr.2019.10410
Pages: 1781-1789
Copyright: © Ma et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

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.

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