Journal: Journal of Cellular and Molecular Medicine

Article Title: SFRP2 Potentiates Metastasis of Colon Cancer by Enhancing Snai1 Protein Stability via USP11

doi: 10.1111/jcmm.71318

Figure Lengend Snippet: SFRP2 is increased in metastatic colon adenocarcinoma and TGF‐beta induced colon cancer cells. (A) Volcano plot was with differentially expressed genes between metastatic colon cancer and primary cancer in GSE40367 . (B) Heatmap showing expression levels of eight up‐regulated and eight down‐regulated different expressed genes in metastatic colon cancer compared to primary colon cancer. (C) Expression of SFRP2 was increased in colon cancer compared to normal tissues in TCGA. (D‐F) Kaplan–Meier plotter of SFRP2 in overall survival (OS), recurrence‐free survival (RFS) and post‐progression survival (PPS) for colon cancer. (G) Representative images of immunohistochemical analysis for SFRP2 in colon cancer samples. IHC score of SFRP2 in normal, primary and metastasis colon cancer tissues. (H) Western blot analysis of the expression of SFRP2 in normal, primary and metastasis colon cancer tissues. (I and J) Western blot analysis of expression of SFRP2, E‐cadherin, N‐cadherin and Vimentin in HCT116 and LoVo cells with different concentrations of TGF‐β1 (0, 10, 20, 50 ng/mL) treatment. *** p < 0.001.

Article Snippet: The membranes were incubated with primary antibodies against E‐cadherin (1:1000, ABclonal, A3044), N‐cadherin (1:1000, ABclonal, A0433), Vimentin (1:1000, ABclonal, A19607), SFRP2 (1:1000, ABclonal, A5383), PI3K (1:1000, CST, #13,666), p‐PI3K (1:1000, CST, #17,366), Akt (1:1000, CST, #4685), p‐Akt (1:1000, CST, #4060), mTOR (1:1000, CST, #2983), p‐mTOR (1:1000, CST, #2971), GAPDH (1:2000, ABclonal, A19056) overnight at 4°C, followed by the α‐mouse or α‐rabbit‐HRP secondary antibodies.

Techniques: Expressing, Immunohistochemical staining, Western Blot

Journal: Journal of Cellular and Molecular Medicine

Article Title: SFRP2 Potentiates Metastasis of Colon Cancer by Enhancing Snai1 Protein Stability via USP11

doi: 10.1111/jcmm.71318

Figure Lengend Snippet: SFRP2 promotes migration and invasion but not proliferation in colon cancer cells. (A) Western blot analysis of SFRP2 in HCT116 cells transfected with SFRP2‐overexpression plasmid and LoVo cells transfected with shSFRP2s. (B) Effects of SFRP2 on proliferation of HCT116 and LoVo cells by CCK‐8 assays. (C) Effects of SFRP2 on migration of HCT116 and LoVo cells by Transwell assays. (D) Effects of SFRP2 on invasion of HCT116 and LoVo cells by Transwell‐Matrigel assays. (E) Epithelial‐mesenchymal transition related proteins such as E‐cadherin, N‐cadherin, Vimentin and SFRP2 were assessed in SFRP2‐overexpression and knockdown cell lines. *** p < 0.001.

Article Snippet: The membranes were incubated with primary antibodies against E‐cadherin (1:1000, ABclonal, A3044), N‐cadherin (1:1000, ABclonal, A0433), Vimentin (1:1000, ABclonal, A19607), SFRP2 (1:1000, ABclonal, A5383), PI3K (1:1000, CST, #13,666), p‐PI3K (1:1000, CST, #17,366), Akt (1:1000, CST, #4685), p‐Akt (1:1000, CST, #4060), mTOR (1:1000, CST, #2983), p‐mTOR (1:1000, CST, #2971), GAPDH (1:2000, ABclonal, A19056) overnight at 4°C, followed by the α‐mouse or α‐rabbit‐HRP secondary antibodies.

Techniques: Migration, Western Blot, Transfection, Over Expression, Plasmid Preparation, CCK-8 Assay, Knockdown

Journal: Journal of Cellular and Molecular Medicine

Article Title: SFRP2 Potentiates Metastasis of Colon Cancer by Enhancing Snai1 Protein Stability via USP11

doi: 10.1111/jcmm.71318

Figure Lengend Snippet: The SFRP2‐Snai1 axis promotes PI3K/AKT/mTOR pathway in colon cancer cells. (A) Gene set enrichment analysis of SFRP2 in PI3K/Akt signalling pathway. (B) Western blot analysis of PI3K, p‐PI3K, Akt, p‐Akt, mTOR, p‐mTOR in HCT116‐vector and HCT116‐SFRP2 treated with shSnai1 or shNC or LoVo‐shNC and LoVo‐shSFRP2 treated with transient transfection of Snai1 overexpression plasmids. (C and D) Effects of SFRP2‐Snai1 on migration of HCT116 and LoVo cells by Transwell assays. (E and F) Effects of SFRP2‐Snai1 on invasion of HCT116 and LoVo cells by Transwell‐matrigel assays. (G and H) Effects of SFRP2‐Snai1 on EMT‐related proteins such as E‐cadherin, N‐cadherin, Vimentin were assessed by western blot. *** p < 0.001.

Article Snippet: The membranes were incubated with primary antibodies against E‐cadherin (1:1000, ABclonal, A3044), N‐cadherin (1:1000, ABclonal, A0433), Vimentin (1:1000, ABclonal, A19607), SFRP2 (1:1000, ABclonal, A5383), PI3K (1:1000, CST, #13,666), p‐PI3K (1:1000, CST, #17,366), Akt (1:1000, CST, #4685), p‐Akt (1:1000, CST, #4060), mTOR (1:1000, CST, #2983), p‐mTOR (1:1000, CST, #2971), GAPDH (1:2000, ABclonal, A19056) overnight at 4°C, followed by the α‐mouse or α‐rabbit‐HRP secondary antibodies.

Techniques: Western Blot, Plasmid Preparation, Transfection, Over Expression, Migration