Nodal Enhances Perineural Invasion in Pancreatic Cancer by Promoting Tumor-Nerve Convergence

Figure 2

Nodal increases the expression of neurotrophins and enhances pancreatic cancer cell invasion. (a) Panc-1 and BxPC-3 cells were incubated with different concentrations of rhNodal (0, 50, and 100 μM) for 24 h and western blot was used to evaluate NGF, BDNF, GDNF, and MMP-9 protein expression. (b) Panc-1 and BxPC-3 cells were pretreated with rhNodal (100 μM) for 24 h and immunofluorescence analysis was conducted to assess the expression of NGF in Panc-1 and BxPC-3 cells. Magnification, ×400. (c) Panc-1 was treated with conditioned medium with different concentrations of rhNodal (0, 50, and 100 μM) for 24 h; then, the supernatant of Panc-1 was collected and an ELISA was performed to evaluate the NGF level. (d) The effect of rhNodal on the invasive ability of Panc-1 and BxPC-3 cells was evaluated by the Matrigel invasion assay. Invasive cells were counted and plotted. Magnification, ×100. .

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