RLN2 Is a Positive Regulator of AKT-2-Induced Gene Expression Required for Osteosarcoma Cells Invasion and Chemoresistance

Figure 7

Effect of RLN2 on invasion and angiogenesis in OS cells. MG-63 cells were transfected with RLN2 siRNA2 for 48 hs and then transfected with myr-AKT (10 μM) for 24 hs or treated with 20 ng/mL of TNF-α for 6 hs; the invasive ability of MG-63 cells was detected by Matrigel invasion assay (a). Quantitative analysis of the tube formation by HUVECs induced by conditioned medium (b). The reduced invasion and tube formation induced by the conditioned medium of RLN2-siRNA2 transfected MG-63 cells were rescued by adding myr-AKT or TNF-α. U-2OS cells were treated with NF-κB inhibitor BAY 11-7082 (10 μM) for 24 hs, or 50 μM LY294002 for 3 hs, or IκBαM (100 nM) for 24 hs; then, the cells were treated with recombinant relaxin for 24 hs; the invasive ability of U-2OS cells was detected by Matrigel invasion assay (c). Quantitative analysis of the tube formation by HUVECs induced by conditioned medium (d). The increased invasion and tube formation induced by the conditioned medium of RLN2 treated U-2OS cells were rescued by adding BAY 11-7082 or LY294002 or IκBαM. .

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