Journal: Cancer Biology & Therapy

Article Title: Fibroblast-associated TPM2 links cell-matrix remodeling to EMT-Notch signaling and gemcitabine resistance in intrahepatic cholangiocarcinoma

doi: 10.1080/15384047.2026.2725344

Figure Lengend Snippet: TPM2 depletion suppresses EMT/ECM-related programs and Notch signaling in intrahepatic cholangiocarcinoma organoids. (A) Representative immunofluorescence staining of CK19, TPM2, and Vimentin in control and TPM2-knockout ICCO2 organoids. CK19 marks cholangiocytic epithelial tumor cells, TPM2 denotes the knockout target, and Vimentin serves as an EMT-associated marker. Nuclei were counterstained with DAPI. Scale bar, 20 μm. (B, C) Quantification of TPM2-positive area (B) and Vimentin-positive area (C) in control and TPM2-knockout ICCO2 organoids. At least 20 fields of view per group were quantified in each independent biological experiment. Positive areas were quantified within organoid regions using ImageJ under identical imaging and thresholding settings. Data are presented as mean ± SEM. *** P < 0.001. (D) RT-qPCR analysis of Notch pathway genes, EMT-associated markers, and ECM-related genes in control and TPM2-knockout ICCO2 organoids. Relative mRNA expression was normalized to GAPDH and is shown relative to the control group. Experiments were performed using three independent biological replicates, with three technical replicates per biological replicate. (E) Representative western blot analysis showing reduced Notch1 and HES1 expression after TPM2 knockout in ICCO2 organoids. β -Tubulin was used as a loading control. Data are representative of three independent biological experiments. (F) Proposed model illustrating the convergence of fibroblast-associated and tumor-cell-intrinsic TPM2 programs on cell-matrix/cytoskeletal remodeling, Notch-EMT signaling, and gemcitabine tolerance. Solid arrows indicate relationships directly supported by the experimental data in the corresponding context, whereas dashed arrows indicate inferred, indirect, or biologically plausible relationships that require further validation.

Article Snippet: Primary antibodies included AFP (#14550-1-AP; Proteintech; 1:200), CK19 (#GB15198; Servicebio; 1:600), ALB (# GB152080 ; Servicebio; 1:50), and Vimentin (#A11952SP; Abclonal; 1:100).

Techniques: Organoids, Immunofluorescence, Staining, Control, Knock-Out, Marker, Imaging, Quantitative RT-PCR, Analysis, Expressing, Western Blot, Cytoskeletal, Biomarker Discovery

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: SFRP2 interacts with Snai1 protein to improve the stability of Snai1 protein. (A) GSEA analysis of SFRP2 co‐upregulated proteins. (B) Correlation analysis of SFRP2 and Snai1 in TCGA colon cancer database. (C) Western blot analysis of Snai1 expression in SFRP2‐overexpression and knockdown cell lines. (D) Endogenous immunoprecipitation of the Snai1 protein by an anti‐SFRP2 antibody in LoVo cells. (E) Immunoprecipitation of the Snai1 protein by an anti‐Flag antibody in HCT116 cells transfected with Flag‐SFRP2. Cells transfected with empty vector were used as a negative control. (F) HCT116 cells were transfected with Flag‐tagged SFRP2 expression plasmids and Myc‐tagged Snai1 expression plasmids then treated with CHX at the concentration of 200 μg/mL at 4, 8, 12 h. The protein degradation of Snai1 was assessed by western blot. (G) Western blot analysis of Snai1 expression in LoVo cells transfected with shSFRP2s with the treatment of MG132 at a concentration of 20 μM for 4 h. ** p < 0.01, *** 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, Expressing, Over Expression, Knockdown, Immunoprecipitation, Transfection, Plasmid Preparation, Negative Control, Concentration Assay

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 relieves Snai1 ubiquitination through deubiquitinase USP11. (A) Correlation analysis of SFRP2 and Snai1 in TCGA colon cancer database. (B) Immunoprecipitation of USP11 protein using anti‐HA antibody in HCT116 cells transfected with HA‐tagged Snai1 and Flag‐tagged USP11 plasmids. (C) Western blot analysis of SFRP2, Snai1 and USP11 after co‐immunoprecipitation in cells co‐transfected with HA‐tagged Snai1, Myc‐tagged SFRP2 and Flag‐tagged USP11. (D) Immunoprecipitation of USP11 using anti‐Snai1 antibody in LoVo cells transfected with shSFRP2s. (E) HCT116‐Vector/HCT116‐SFRP2 cells were transfected with vector, Flag‐USP11, Flag‐USP11 with treatment of USP11 inhibitor mitoxantrone (5 nmol/L). Western blot analysis of SFRP2, Snai1 and USP11 was measured in the above groups. (F) Western blot analysis of Snai1 expression in SFRP2‐knockdown LoVo cells transfected with vector or Flag‐USP11. GAPDH was used as a loading control.

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: Ubiquitin Proteomics, Immunoprecipitation, Transfection, Western Blot, Co-Immunoprecipitation Assay, Plasmid Preparation, Expressing, Knockdown, Control

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: Snai1 directly bound to the SFRP2 promoter and increased SFRP2 transcription. (A) The predicted binding site of Snail1 in the promoter of SFRP2. (B‐C) Western blot analysis of SFRP2 in Snai1‐expression HCT116 cells and Snai1‐knockdown LoVo cells. (D) Luciferase reporter analysis of SFRP2‐promoter in Snai1‐expression HCT116 cells. (E) Luciferase reporter analysis of SFRP2‐promoter in Snai1‐knockdown LoVo cells. (F) The schematic of luciferase reporter plasmids of SFRP2 wildtype promoter and SFRP2 mutant promoter. (G) Luciferase reporter analysis of SFRP2‐promoter and SFRP2‐mutant‐promoter in HCT116 cells. (H) Agarose electrophoresis for ChIP analysis of Snai1 binding to the SFRP2 promoter. (I) qRT–PCR for ChIP analysis of Snai1 binding to the SFRP2 promoter. *** 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: Binding Assay, Western Blot, Expressing, Knockdown, Luciferase, Mutagenesis, Electrophoresis, Quantitative RT-PCR

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

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 metastasis in vivo. (A) Representative bioluminescence images of NSG mice injected with SFRP2‐expression with or without Snai1‐knockdown stable HCT116 cells. (B) Western blot analysis of expression of PI3K, p‐PI3K, Akt, p‐Akt, mTOR, p‐mTOR, Snai1, SFRP2 of 3 groups of tumour‐bearing mice. (C) Representative bioluminescence images of NSG mice injected with SFRP2‐knockdown with or without Snai1 expression stable LoVo cells. (D) Western blot analysis of expression of PI3K, p‐PI3K, Akt, p‐Akt, mTOR, p‐mTOR, Snai1, SFRP2 of the above 3 groups of tumour‐bearing mice. (E) The schematic illustration of SFRP2/Snai1/USP11 positive feedback loop promoting colon cancer metastasis via the PI3K/Akt/mTOR pathway. *** 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: In Vivo, Injection, Expressing, Knockdown, Western Blot

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 drives tumor-educated M2-like polarization of macrophages and promotes tumor-cell aggressiveness. (A) Workflow for generating TC-hMDMs and TC-BMDMs, circSMAD4 knockdown, and downstream functional assays. (B) RT–qPCR analysis of M1-associated markers (MHC-II [HLA-DRA in TC-hMDMs; H2-Ab1 in TC-BMDMs], NOS2, and CD86) and M2-associated markers (CD163, CD206, and ARG1) in TC-hMDMs and TC-BMDMs. (C) Representative flow-cytometry histograms for HLA-DR, iNOS, CD86, CD163, CD206, and ARG1 in TC-hMDMs. Gating strategy and marker thresholds were defined based on FMO controls (see ). (D) Flow-cytometry quantification of marker-positive cells in TC-hMDMs and TC-BMDMs. (E) ELISA of IL-10, TGF-β, and iNOS in culture supernatants. (F) CCK-8 assays of A549 and LLC cells. (G) Colony-formation assays of A549 and LLC cells with quantification. (H) Bioluminescence-based growth readouts of patient-derived LUAD organoids (PDO #1 and PDO #2) after co-culture with TC-hMDMs. (I) Immunoblot analysis of EMT-related proteins (E-cadherin, N-cadherin, Vimentin) in A549 and LLC cells. (J) Transwell migration and invasion assays of A549 and LLC cells with quantification. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: Knockdown, Functional Assay, Quantitative RT-PCR, Flow Cytometry, Marker, Enzyme-linked Immunosorbent Assay, CCK-8 Assay, Derivative Assay, Co-Culture Assay, Western Blot, Migration

Journal: Non-coding RNA Research

Article Title: CircSMAD4 shapes matrix-remodeling TAMs in lung adenocarcinoma

doi: 10.1016/j.ncrna.2026.03.003

Figure Lengend Snippet: circSMAD4 depletion in macrophages restrains LUAD growth and metastasis in vivo. (A) Schematic of orthotopic lung implantation and experimental metastasis models using LLC cells mixed with BMDMs expressing shNC or sh-circSMAD4. (B) Representative images of orthotopic lung tumors. (C) Tumor weight of orthotopic implants. (D) Overall survival of mice bearing orthotopic tumors. (E) Immunofluorescence showing F4/80 and circSMAD4 signals in tumor tissues. Scale bar, 50 μm. (F, G) Representative Ki-67 IHC staining and quantification in orthotopic tumors. Scale bar, 50 μm. (H) Representative bioluminescence images of lung tumor burden in the metastasis model. (I) Tumor weight in the metastasis model. (J) Overall survival of mice in the metastasis model. (K–M) Representative IHC staining and quantification of E-cadherin and vimentin in tumors. Scale bar, 50 μm. ∗P < 0.05; ∗∗P < 0.01; ∗∗∗P < 0.001; ∗∗∗∗P < 0.0001; ns, not significant.

Article Snippet: Sections were incubated with primary antibodies against Ki-67 (Servicebio, Cat# GB111499 ), E-cadherin (Proteintech, Cat# 20874-1-AP), and Vimentin (Proteintech, Cat# 10366-1-AP).

Techniques: In Vivo, Expressing, Immunofluorescence, Immunohistochemistry