Long non-coding RNA GASL1 promotes proliferation of vascular smooth muscle cells through Wnt/β-catenin signaling pathway

Abstract

Objective To investigate whether long non-coding (lnc) RNA GASL1 can promote the proliferation of vascular smooth muscle cells (VSMCs) by activating the Wnt/β-catenin signaling pathway. Methods The mRNAs co-expression network related to lncRNA GASL1 was constructed and analyzed with Pathway analysis to predict the signaling pathway of the highest correlation with lncRNA GasL1. After LncRNA GASL1 and lncRNA negative control (lncRNA NC) were transfected into VSMCs respectively, cell counting kit-8 (CCK-8) assay was performed to detect VSMCs viability. Immunofluorescence assay was performed to determine the expression level of proliferation-related protein Bcl-2, and Western blotting to detect the levels of the proteins of Wnt/β-catenin signaling pathway. Finally, Wnt/β-catenin pathway inhibitor ICG001 was added into VSMCs along with lncRNA GASL1 for co-culturing (lncRNA GASL1+ ICG001 group), and the alterations of VSMCs viability and bcl-2 protein level were detected and compared with those in the lncRNA GASL1+ control group. Results There were 374 mRNAs associated with lncRNA GASL1, of which the Wnt/β-catenin pathway had the highest correlation. CCK-8 assay showed that the VSMCs viability was higher in the lncRNA GASL1 group than in the lncRNA NC group (P < 0.05). The results of immunofluorescence assay and Western blotting revealed that both the Bcl-2 protein level and the levels of Wnt1 and β-catenin were increased in lncRNA GASL1 group (P < 0.05) when compared to the lncRNA-NC group. After co-culturing with lncRNA GASL1 and ICG001, the VSMCs viability was significantly decreased (P < 0.05), and the Bcl-2 protein level was decreased when compared with the the lncRNA GASL1+control group (P < 0.05). Conclusion LncRNA GASL1 promotes the proliferation of VSMCs through the Wnt/β-catenin signaling pathway. Blocking the signaling pathway can reduce the promotive effect of lncRNA GASL1 on VSMCs.

Keywords

• lncrna gasl1
• cell proliferation
• vascular smooth muscle cells
• wnt/β-catenin signaling pathway