Stromal fibroblast activation protein alpha promotes gastric cancer progression via epithelial-mesenchymal transition through Wnt/ β-catenin pathway

Figures

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  Fig. 1. Correlations between FAP and OS of GC patients. FAP was expressed both in GC cells (a) and stromal CAFs (b, shown by red arrowheads). c FAP was not expressed in normal tissues. d The median OS of GC cases with high expression of FAP (30.2 months) was shorter than that with low expression of FAP (37.8 months), the difference was statically different (P < 0.01)

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  Fig. 2. Stromal FAP promotes the proliferation, migration, and invasion abilities of SGC7901. In advance, SGC-7901 cells had been cocultured with HELF (SH group), HELFNC cells (SN group) and HELFFAP cells (SF group) for 72 h, respectively. a Exogenous FAP promotes SGC7901 proliferation in dose-dependent manner. b Exogenous FAP promotes SGC7901 migration in dose-dependent manner. c Exogenous FAP promotes SGC7901 invasion in dose-dependent manner. d Cell viability was determined by CCK8 assay. SGC7901 cells in SF group were promoted to proliferate. e Clone formation assay of SGC7901 cells in SF and SN groups. f Western blot assay indicated that the expression of PCNA and MMP9 in SF group was highest. g Wound healing assay indicated that the width of injury was lower in SF group in 24 h. h Cell migration and invasion abilities were determined by Transwell assay. The number of migrated and invasive SGC7901 cells in SF group was much higher than that in SN group

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  Fig. 3. FAP inhibits the apoptosis of SGC7901 cells. a Apoptotic SGC7901 cells in NC group was higher than that in FAP group, whereas the result turned to the opposite in GES1 cells. b The cellular circle of SGC7901 was also detected by FCM. No differences could be observed in FAP and NC groups

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  Fig. 4. FAP promotes EMT of GC cells through Wnt/β-catenin pathway. Exogenous FAP promotes SGC EMT in dose-dependent manner. The expression of E-cadherin and ZO-1 were reduced, while that of N-cadherin and Vimentin were increased by qRT-PCR assay (a), and Western blotting assay (b). In addition, the DKK1 and LEF-1 protein, which could be participated in Wnt/β-catenin pathway, were also increased with more exogenous FAP. The result also accompanied in SGC cells co-cultured in SN and SF groups. c The morphology of SGC cells in SF group tended to be fibroblast-like, long fusiform, which was indicated by red arrows. d The expression of E-cadherin and ZO-1 were reduced, while that of N-cadherin and Vimentin were increased in SGC cells of SF group by Western blotting assay. Similarly, the DKK1 and LEF-1 protein were also increased. e The expression of E-cadherin was reduced, while that of α-SMA was increased both in GES1 cells (gastric normal cells) and SGC cells of SF group by the immunofluorescence staining

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  Fig. 5. FAP promotes GC progression in a xenograft gastric cancer nude mouse model. a The combination of SGC7901 and HELFFAP was much more effective in elevating tumor burden. The tumor volume (b) and weight (c) in the NC group were significantly lower than FAP group. d Representative immunohistochemical analysis of CD31, Ki67 (200× magnifications, Scale bar 50 μm)