WISP1 overexpression promotes proliferation and migration of human vascular smooth muscle cells via AKT signaling pathway.
Proliferation and migration of vascular smooth muscle cells (VSMCs) play crucial roles in the development of vascular restenosis. Our previous study showed that CCN4, namely Wnt1 inducible signaling pathway protein 1 (WISP1), significantly promotes proliferation and migration of rat VSMCs, but its mechanism remains unclear. This study aims to investigate whether and how WISP1 stimulates proliferation and migration of human VSMCs. Western blot analysis showed that FBS treatment increased WISP1 protein levels in human VSMCs in a dose-dependent manner. Overexpression of WISP1 using adenovirus encoding WISP1 (AD-WISP1) significantly increased proliferation rate of human VSMCs by 2.98-fold compared with empty virus (EV)-transfected cells, shown by EdU incorporation assay. Additionally, Scratch-induced wound healing assay revealed that adenovirus-mediated overexpression of WISP1 significantly increased cell migration compared with EV-transfected cells from 6h (4.56±1.14% vs. 11.23±2.25%, P<0.05) to 48h (25.25±5.51% vs. 97.54±13.12%, P<0.01) after injury. Transwell Migration Assay confirmed that WISP1 overexpression significantly promoted human VSMC migration by 2.25-fold compared with EV. Furthermore, WISP1 overexpression stimulated Akt signaling activation in human VSMCs. Blockage of Akt signaling by Akt inhibitor AZD5363 or PI3K inhibitor LY294002, led to an inhibitory effect of WISP1-induced proliferation and migration in human VSMCs. Moreover, we found that WISP1 overexpression stimulated GSK3α/β phosphorylation, and increased expression of cyclin D1 and MMP9 in human VSMCs, and this effect was abolished by AZD5363. Collectively, we demonstrated that Akt signaling pathway mediates WISP1-induced migration and proliferation of human VSMCs, suggesting that WISP1 may act as a novel potential therapeutic target for vascular restenosis.
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China.
Shun
S
Lu
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China.
Hao
H
Liu
Department of Pathophysiology, Zhongshan Medical School, Sun Yat-Sen University, China.
Lihe
L
Lu
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China.
Heng
H
Wan
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China.
Zhiqi
Z
Lin
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China.
Kai
K
Qian
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China.
Xingxing
X
Yao
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China.
Qing
Q
Chen
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China.
Wenjun
W
Liu
Department of Histology and Embryology, Key Laboratory of Tissue Construction and Detection of Guangdong Province, Southern Medical University, China. Electronic address: yanjy790@163.com.
Jianyun
J
Yan
Department of Vascular Surgery, Nanfang Hospital, Southern Medical University, China. Electronic address: zhengjunliugz@163.com.
Zhengjun
Z
Liu
D055511:CCN Intercellular Signaling Proteins / Q000235:genetics*; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation / Q000235:genetics; D019938:Cyclin D1 / Q000378:metabolism; D015870:Gene Expression; D000071679:Glycogen Synthase Kinase 3 beta / Q000378:metabolism; D006801:Humans; D020780:Matrix Metalloproteinase 9 / Q000378:metabolism; D009131:Muscle, Smooth, Vascular / Q000166:cytology*; D019869:Phosphatidylinositol 3-Kinases / Q000378:metabolism; D010766:Phosphorylation / Q000235:genetics; D011518:Proto-Oncogene Proteins / Q000235:genetics*; D051057:Proto-Oncogene Proteins c-akt / Q000378:metabolism*; D015398:Signal Transduction / Q000235:genetics; D015854:Up-Regulation
D055511:CCN Intercellular Signaling Proteins; C116396:CCN4 protein, human; D011518:Proto-Oncogene Proteins; D019938:Cyclin D1; D019869:Phosphatidylinositol 3-Kinases; D000071679:Glycogen Synthase Kinase 3 beta; D051057:Proto-Oncogene Proteins c-akt; D020780:Matrix Metalloproteinase 9
Migration; Proliferation; Restenosis; Vascular smooth muscle cells; WISP1