1. Hoxa9 is partly required for Twist1-induced migration, invasion, anoikis resistance, and soft agar tumorigenicity of Myc-CaP cells.
Hoxa9 is partly required for Twist1-induced migration, invasion, anoikis resistance, and soft agar tumorigenicity of Myc-CaP cells. A, Myc-CaP cell stably overexpressing Twist1 were transduced with shRNA constructs against Hoxa9 or scrambled control and relative expression determined by Hoxa9 qPCR (left, n ≥ 3) and Hoxa9 Western blotting (right). B, scratch wound-healing assay of Myc-CaP + Twist1 isogenic cell lines and representative images shown at 0hour and 48 hours. C, in relative wound closure calculations (as in Fig. 3; all n = 3, 3 fields per experiment). D, Transwell invasion assays with Matrigel were conducted with isogenic Myc-CaP + Twist1 cells (n = 6, represented by column mean ±SEM). E, cells were grown adherent or in suspension using ultra-low attachment dishes and anoikis quantified by AnnexinV- AlexaFluor 488 and propidium iodide staining (as in Fig. 4) plotted as bar graphs ±SEM. A total of 5 × 105 cells were embedded in soft agar and incubated for 2 weeks. Colonies containing above 50 cells were scored in at least 5 random fields. F, representative phase contrast images ×40 magnifications are shown. G, the percent clonogenicity in soft agar is calculated by normalizing the number of colonies to the total number of cells and represented as bar graphs ±SEM (n ≥ 4). All comparisons, *, P < 0.05; **, P < 0.01; and ***, P < by Mann–Whitney test.
Rajendra P. Gajula et al. Mol Cancer Res 2013;11:
©2013 by American Association for Cancer Research