Figure 2
From: GATA3 interacts with and stabilizes HIF-1α to enhance cancer cell invasiveness
GATA3 promotes tumour invasiveness in vitro and in vivo. (a) Western blot analysis of GATA3 expression in various cell lines as indicated. GAPDH was an internal control. (b) Knockdown and overexpression of GATA3 were confirmed by western blot analysis. For knockdown, OEC-M1 cells were transfected with non-targeting siRNA (siCtr) or two independent siRNAs against GATA3 (siGATA3-1 and siGATA3-2). For overexpression, FaDu and A375 cells were transfected with empty pcDNA3.1 (Mock) or GATA3/pcDNA3.1 (GATA3). (c) Effects of GATA3 on cell migration and invasion analysed by transwell migration and Matrigel invasion assays, respectively. Cells were seeded after transfection for 48 h. Normoxia (N, O2=21%) or hypoxia (H, O2=1%) is as indicated. Results are represented as mean±s.d. from three independent experiments. *P<0.05, **P<0.01. (d) Effects of GATA3 on experimental metastasis in vivo. Upper panel, representative images of lungs and metastatic tumour nodules indicated by arrowheads. Lower panel, statistical analysis of tumour nodules. Mock or GATA3 overexpressing OEC-M1 and A375 stable transfectants (1 × 106 cells / per injection) were injected through tail veins into NOD-SCID mice (n=6 and 5, respectively). Results are represented as mean±s.d. Data are analysed by Student’s t-test. *P<0.05. (e) Expression of GATA3 and HIF-1α in subcutaneous tumour xenografts. Left, hematoxylin and eosin staining. The hollow arrow indicates the intact capsule separating the tumour from surrounding tissues. The black arrow indicates the invasion of tumour into skeletal muscles. M, skeletal muscle. T, tumour. Middle and right, immunohistochemical staining of GATA3 and HIF-1α. Scale bars, 50 μm.
