Fig 2: HDAC7 deacetylated β-catenin at Lys49 and promoted its translocation into the nucleus to activate FGF18 expression. a HDAC7 and β-catenin interacted in NSCLC cells. b Two-color immunofluorescence analysis showing significant colocalization of HDAC7 with β-catenin in A549 and SK-LU-1 cells. DAPI (blue), HDAC7 (green), and β-catenin (red). c Representative western blotting results showing HDAC7 deacetylated β-catenin at Lys49 and phosphorylated β-catenin at Ser45. d Representative western blotting showing upregulated HDAC7 facilitated β-catenin nuclear translocation. e Representative western blotting of HDAC7, FGF18, β-catenin, and TCF4 in NSCLC cells after LF3 treatment. β-actin was used as an internal control to normalize the total protein amount. GAPDH and Lamin B1 were used as the loading control to cytosolic protein and nuclear protein, respectively. DAPI fluorescence marking the cell nucleus. LV, lentivirus. IP, immunoprecipitation. IB, immunoblot

Fig 3: HDAC7 overexpression facilitated NSCLC metastasis in vitro and vivo. a Representative scratch wound assay images and results. Migration ability is expressed as the mean scratch area at each time point. The initial scratched area (0 h) was set as 100%. b Representative transwell migration assay images and results. Representative western blotting of HDAC7, E-cadherin, and Snail in HDAC7 overexpression and knockdown cells (c) and xenograft tumor tissues (d). Representative fluorescence images (e) and fluorescence signals analysis (f) of pulmonary metastases 4 weeks after tail vein injection. g Representative HE staining images of lung samples from indicated groups. Scale bar, 1 mm and 100 μm (inset), respectively. h The number of metastatic nodules per lung in HE staining images. i Representative general morphology of surface lung metastases. The black triangular arrows indicate the metastatic nodules. β-actin was used as internal control. All the data are expressed as mean ± SD. *P < 0.05. LV, lentivirus

Fig 4: TMP269 attenuates WNT5A-mediated ESCC cell migration and invasion.A Representative western blots showing the levels of FLAG-WNT5A, HDAC7, SNAIL, E-cadherin, and vimentin in WNT5A-overexpressing EC109 and KYSE450 cells after treatment with TMP269 for 48 h. β-actin was used as an internal control. B, C Representative images and statistical analysis of wound healing assays and Transwell migration/invasion assays using WNT5A-overexpressing ESCC cells after treatment with TMP269 (20 μM) for 48 h. The migratory ability in wound healing assays was expressed as the mean scratch area. The initial scratch area (0 h) was set to 100%. *P < 0.05 compared with the LV-Control group, #P < 0.05 compared with the LV-Control + TMP269 group, and &P < 0.05 compared with the LV-WNT5A group. LV lentivirus. D Schematic diagram of the mechanism by which WNT5A promotes tumor metastasis by activating the HDAC7/SNAIL signaling pathway in ESCC.

Fig 5: The correlation of HDAC7 and FGF18 expression in NSCLC. a Kaplan–Meier survival analysis of FGF18 expression in 1925 NSCLC cases from the Kaplan–Meier plotter database. b Western blot analysis of FGF18 expression in tumor (T) and paired adjacent nontumor (N) tissues from 12 NSCLC patients. (c) Representative FGF18 IHC images of NSCLC and adjacent non-tumor tissues. Scale bar, 200 μm and 20 μm (inset), respectively. d The IHC analysis of FGF18 expression in 319 NSCLC patients. e Kaplan–Meier survival analysis of FGF18 expression in 319 NSCLC patients. f Correlation analysis of HDAC7 and FGF18 expression in NSCLC tissues. g The comprehensive Kaplan–Meier survival analysis of HDAC7 and FGF18 in 319 NSCLC patients based on the microarray tissue IHC results. h The schematic of a simplified regulatory network involving USP10 and FGF18 in HDAC7-meditated NSCLC progression. USP10 interacts with and stabilizes HDAC7 through deubiquitination, preventing its proteasomal degradation. Elevated HDAC7 expression facilitates β-catenin redistribution to the nucleus by deacetylating β-catenin and inhibiting its phosphorylation. As a result, the nuclear accumulation of β-catenin binds to TCF/LEF to activate the transcription of FGF18 and thus promotes NSCLC proliferation and metastasis