Ovarian BDNF promotes survival migration and attachment of tumor precursors originated from p53 mutant fallopian tube epithelial cells

摘要

Partial sequences of wild-type p53 and shRNA-resistant p53 mutants. The graph was based on the plasmid DNA sequencing result. Representative images of western blot. The whole-cell lysate of FT240 cells overexpressing green fluorescent protein (GFP), p53R273H, p53R175H, or p53R248W were analyzed. , Band density quantification of p53 and TrkB western blot. Band density was normalized to the data of β-actin control and compared to the control FT240 expressing GFP.  = 3. *  p #  e Flow cytometry of TrkB in human fallopian tube cell lines FT240, FT246, and FT340. 2D and 3D indicate the regular 2D adhesion culture and cell suspension 3D culture, respectively. *  t test. BDNF suppressed Caspase3/7 activity of FT240 cells in 3D culture. Data were normalized to Caspase3/7 activity of untreated cells in 2D culture (NT 2D).  = 3. **  p #  g BDNF (50 ng/ml) promoted the recovery of FT240 cells (FTEs) from anoikis-inducing condition. Cell viability was quantified after FTEs reattached to collagen I-coated matrix for 48 h. DAPI staining was used to visualize the nuclei (as white dots) of the recovered cells in the representative images. Cell viability was determined using CellTiter-Glo 2D Cell Viability Assay.  = 8. **  t test. BDNF (50 ng/ml) accelerated the migration of FTEs from hydrogel. Migrated cells were visualized by crystal violet staining. After hydrogel pieces were removed, the migrated cells were quantified with CellTiter-Glo 2D Cell Viability Assay.  = 3. **  t test. BDNF (50 ng/ml) enhanced the attachment of FTEs to Collagen I-coated beads. Representative images indicate the attachment of red fluorescent FT240 to beads after 24-h incubation as expected. The percentage of attached cells is incubation-time-dependent. BDNF treatment accelerated the attachment. **  p