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FOXQ1 gene mediates TGF-β1 signaling pathway to regulate in vitro angiopoiesis of pancreatic cancer PANC-1 cells
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Abstract:
Objective: To explore the effect of forkhead box Q1（FOXQ1）gene silencing on in vitro angiopoiesis of pancreatic carcinoma cells, and to investigate its role in transforming growth factor-β1(TGF-β1) signaling pathway. Methods: FOXQ1-shRNA recombinant lentiviral vector and negative control lentiviral vector (NC-shRNA) were transfected into PANC-1 cells. Flow cytometry was used to detect the transfection efficiency. The gene silencing efficiency was measured by qPCR and Western blotting. FOXQ1-shRNA group,NC-shRNA group and blank group were set up. Human umbilical vein endothelial cells (HUVECs）were used for the in vitro angiopoiesis assay, which was observed under fluorescence microscopy. Meanwhile, qPCR and Western blotting were performed to examine mRNA and protein expressions of VEGF-A and MMP-2, respectively. After induction by TGF-β1 (final concentration of 5 ng/ml), the changes in angiopoiesis ability as well as the expression changes in FOXQ1，VEGF-A，MMP-2 in each group were detected. Results:The transfection efficiency of lentivirus was about 90%. Compared with NC-shRNA group, the angiopoiesis ability in FOXQ1-shRNA group was remarkably decreased（9.33±2.08 vs 28.67±2.52，P<0.05); Meanwhile, the expressions of VEGF-A and MMP-2 were all declined significantly (P<0.05). TGF-β1 improved the mRNA and protein expressions of FOXQ1，VEGF-A and MMP-2, and increased the in vitro angiopoiesis ability (P<0.05). Conclusion: FOXQ1 gene could mediate the in vitro angiopoiesis of PANC-1 cells; its mechanism may be related to the down-egulation of VEGF-A and MMP-2 and possibly be regulated by TGF-β1 pathway.
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Project Supported:
Project supported by Scientific Research Foundation of the Education Department of Sichuan Province（No.12ZB218）
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