Fig. 3 | Signal Transduction and Targeted Therapy

Fig. 3

From: AKT2S128/CCTαS315/319/323-positive cancer-associated fibroblasts (CAFs) mediate focal adhesion kinase (FAK) inhibitors resistance via secreting phosphatidylcholines (PCs)

Fig. 3

CAFs-released PCs induce the resistance of defactinib. a KYSE410 and KYSE510 cells were treated with 10 μM PC (16:0/20:4) or glycerophospholipid with defactinib (0–10 μM) for 4 days, then the growth of indicated ESCC cells was evaluated using MTS assay. IC50 value of defactinib in KYSE410 and KYSE510 cells was shown. b Transwell apparatus with 0.4 μm pore size was used to evaluate the CAFs-derived CCTα or CCTβ-mediated growth of tumor cells in the presence of defactinib. The control siRNA, CCTα siRNA1/2, or CCTβ siRNA1/2 CAFs #1 were plated in the upper chamber of transwell plates. The KYSE410 or KYSE510 cells were respectively plated in the lower chamber of transwell plates, and cocultured with indicated CAFs #1 with defactinib (0–10 μM) for 4 days, and then growth of indicated ESCC cells was measured using MTS assay. IC50 value of defactinib in KYSE410 and KYSE510 cells was shown. c CAFs #1 were plated in the upper chamber of transwell plates with 0.4 μm pore size. The KYSE410 or KYSE510 cells were respectively plated in the lower chamber of transwell plates, and cocultured with CAFs #1 with miltefosine (25 μM) and defactinib (0–10 μM) for 4 days, and then growth of indicated ESCC cells was measured using MTS assay. IC50 value of defactinib in KYSE410 and KYSE510 cells was shown. d KYSE410 and KYSE510 cells were treated with 10 μM PC (16:0/20:4) or glycerophospholipid in the presence of defactinib (10 μM) for 24 h, and the invasion of indicated ESCC cells was evaluated using Transwell invasion assay. e Indicated CAFs #1 were cultured in Transwell apparatus with 8 μm pore size, KYSE410 (left panel) or KYSE510 (right panel) cells were cultured in the upper chamber of transwell plates and treated with 10 μM defactinib for 24 h. The invasion of indicated ESCC cells was evaluated using Transwell invasion assay. f CAFs #1 was cultured in Transwell apparatus with 8 μm pore size, KYSE410 or KYSE510 cells were cultured in the upper chamber of transwell plates and treated with 10 μM defactinib with/without miltefosine (25 μM) for 24 h. The invasion of indicated ESCC cells was evaluated using Transwell invasion assay. The invasive ratio of defactinib/respective control was listed. n.s. no significant difference; ***P < 0.001. Error bars, mean ± SD of three to five independent experiments

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