Overexpression of the growth arrest-specific homeobox gene Gax inhibits proliferation, migration, cell cycle progression, and apoptosis in serum-induced vascular smooth muscle cells

Abstract

ABSTRACT. The Gax gene has been implicated in a variety of celldevelopmental and biological processes, and aberrant Gax expression is linked to many diseases. In this study, to provide important insights for Gax-based gene therapy in vein graft restenosis and its anti-restenotic mechanism, we used rabbit vascular smooth muscle cells (VSMCs) to investigate the effects of Gax overexpression on proliferation, migration, cell cycle, and apoptosis in a serum-stimulated culture. Rabbit VSMC lines that stably overexpressed Gax were established by transfection with recombinant adenoviral vector Ad5-Gax. The effect of Gax overexpression on in vitro serum-induced VSMCs proliferation, migration, cell cycle, and apoptosis was assessed by MTT, wound healing, and flow cytometry assays, respectively. To investigate the effect of Gax overexpression on PCNA and MMP-2 in serum-induced VSMCs, immunocytochemistry, RT-PCR, and gelatin zymography were performed. The results clearly showed that Gax overexpression decreases PCNA expression in serum-induced VSMCs. Gax overexpression also significantly inhibited cell proliferation by blocking entry into the S-phase of the cell cycle, promoted cell apoptosis, and reduced cell migration activity by downregulating MMP-2 release and activity. These findings indicate that Gax would be an optimal target gene for gene therapy to treat vein graft restenosis.

HHEX: A Crosstalker between HCMV Infection and Proliferation of VSMCs

“…HOX gene, as a transcription factor, can combine with target DNA, regulating downstream gene expression involved in angiogenesis and repair, which closely related to proliferative vascular disease pathogenesis (Zheng et al, 2014). So, whether vessel proliferative disease induced by HCMV is related with HOX genes?…”

“…Research has shown that HOX genes participate in a variety of cell proliferation and migration by direct or indirect way, i.e., over expression of growth arresting-specific homeobox (Gax) inhibits the proliferation and migration VSMCs induced by serum (Zheng et al, 2014). And the expression of HOXC11 can induce cell proliferation in renal cell carcinoma (Liu et al, 2015).…”

Epothilones Suppress Neointimal Thickening in the Rat Carotid Balloon-Injury Model by Inducing Vascular Smooth Muscle Cell Apoptosis through p53-Dependent Signaling Pathway

“…Uncontrolled neointimal tissue accumulation of VSMCs shows some similarity with the tumor cell growth and benign tissue proliferation [ 35 ]. Thus, the regulation of apoptosis has attracted considerable attention as an effective way to eliminate hyper-proliferative VSMCs in neointimal formation [ 11 – 13 , 36 – 38 ]. While investigating the pro-apoptotic effect of EPOs, we initially observed that EPO-B and EPO-D potently inhibited PDGF-BB-induced VSMC proliferation in a dose- and time-dependent manner, consistent with previous studies [ 26 , 27 ].…”

“…Mitotic catastrophe of VSMCs was determined by DAPI staining as previously described [ 11 ]. In brief, cells were cultured on an eight chambered glass culture slide (BD Biosciences) at a density of 1 × 10 4 cells/well and cultured for 24 h, and then cells were serum starved for 12 h prior to the treatment with EPO-B or EPO-D (1 to 100 nM), PTX (100 nM), or DMSO (final concentration of 0.1%) as a vehicle control in the presence of PDGF-BB (50 ng/mL).…”

“…Abnormal tissue growth depends on the delicate balance between cell proliferation and apoptosis. It has been demonstrated that increasing VSMCs apoptosis could decrease neointimal hyperplasia [ 11 , 12 ]. Further, prior efforts to reduce the extent of restenosis have focused on means of reducing the proliferation and migration of VSMCs or of increasing their apoptosis [ 13 ].…”