SPINT2 is involved in the proliferation, migration and phenotypic switching of aortic smooth muscle cells: Implications for the pathogenesis of thoracic aortic dissection

Li,Jun; Yu,Changjun; Yu,Kangmin; Chen,Zhiyong; Xing,Dan; Zha,Binshan; Xie,Wentao; Ouyang,Huan

doi:10.3892/etm.2023.12245

SPINT2 is involved in the proliferation, migration and phenotypic switching of aortic smooth muscle cells: Implications for the pathogenesis of thoracic aortic dissection

Authors:
- Jun Li
- Changjun Yu
- Kangmin Yu
- Zhiyong Chen
- Dan Xing
- Binshan Zha
- Wentao Xie
- Huan Ouyang
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Affiliations: Department of Vascular Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China, Department of Gastrointestinal Surgery, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China, Department of Medical Record Management, The First Affiliated Hospital of Anhui Medical University, Hefei, Anhui 230022, P.R. China
Published online on: October 9, 2023 https://doi.org/10.3892/etm.2023.12245
Article Number: 546
Copyright: © Li et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Thoracic aortic dissection (TAD) is a severe and extremely dangerous cardiovascular disease. Proliferation, migration and phenotypic switching of vascular smooth muscle cells (SMCs) are major pathogenetic mechanisms involved in the development of TAD. The present study was designed to investigate the expression and potential function of serine peptidase inhibitor Kunitz type 2 (SPINT2) in TAD. The gene expression profile data for ascending aorta from patients with TAD were downloaded from the GEO database with the accession number GSE52093. Bioinformatics analysis using GEO2R indicated that the differentially expressed SPINT2 was prominently decreased in TAD. The expression levels of SPINT2 mRNA and protein in aortic dissection specimens and normal aorta tissues were measured using reverse transcription‑quantitative PCR and western blotting. SPINT2 expression was downregulated in clinical samples from aortic dissection specimens of patients with TAD compared with the corresponding expression noted in tissues derived from patients without TAD. In vitro, platelet‑derived growth factor BB (PDGF‑BB) was applied to induce the isolated primary mouse aortic SMC phenotypic modulation (a significant upregulation in the expression levels of synthetic markers), and the SMCs were infected with the adenoviral vector, Ad‑SPINT2, to construct SPINT2‑overexpressed cell lines. SMC viability was detected by an MTT assay and SMC proliferation was detected via the presence of Ki‑67‑positive cells (immunofluorescence staining). To explore the effects of SPINT2 on SMC migration, a wound healing assay was conducted. ELISA and western blotting assays were used to measure the content and expression levels of MMP‑2 and MMP‑9. The expression levels of vimentin, collagen I, α‑SMA and SM22α were measured using western blotting. The PDGF‑BB‑induced proliferation and migration of SMCs were recovered by SPINT2 overexpression. The increase in the expression levels of SPINT2 reduced the expression levels of active matrix metalloproteinases (MMPs), MMP‑2 and MMP‑9. Overexpression of SPINT2 suppressed SMC switching from a contractile to a synthetic type, as evidenced by decreased vimentin and collagen I expression levels along with increased α‑smooth muscle actin and smooth muscle protein 22‑α expression levels. Furthermore, activation of ERK was inhibited in SPINT2‑overexpressing SMCs. A specific ERK agonist, 12‑O‑tetradecanoylphorbol‑13‑acetate, reversed the SPINT2‑mediated inhibition of SMC migration and the phenotypic switching. Collectively, the data indicated that SPINT2 was implicated in the proliferation, migration and phenotypic switching of aortic SMCs, suggesting that it may be involved in TAD progression.

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