Journal: Journal of translational medicine

Article Title: Disheveled3 enhanced EMT and cancer stem-like cells properties via Wnt/β-catenin/c-Myc/SOX2 pathway in colorectal cancer.

doi: 10.1186/s12967-023-04120-8

Figure Lengend Snippet: Fig. 1 DVL3 overexpression was positively associated with poor prognosis in CRC patients. A–D Data from the UALCAN database showed that DVL3 was elevated in CRC tissues compared to normal controls. E The result of western blotting revealed that DVL3 expression was stronger in HT-29, HCT-8, SW480, SW620 and HCT116 cells than that in FHC cells. F DVL3 was upregulated in N1 and N2 stage of CRC nodal metastasis compared to N0 stage in data from the UALCAN database. G CRC patients with high DVL3 expression presented shorter overall survival (OS), H, I disease free Survival (DFS), J disease-specific survival (DSS) than patients with Low DVL3 expression, based on data from PrognoScan database. *P ≤ 0.05

Article Snippet: Primary antibodies against DVL3 and β-catenin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Over Expression, Western Blot, Expressing

Journal: Journal of translational medicine

Article Title: Disheveled3 enhanced EMT and cancer stem-like cells properties via Wnt/β-catenin/c-Myc/SOX2 pathway in colorectal cancer.

doi: 10.1186/s12967-023-04120-8

Figure Lengend Snippet: Fig. 2 DVL3 enhanced metastatic potential of CRC cells. Transwell assays were used to determine the potential migration and invasion of HCT-8 (A) and SW620 (B) cells transfected with pcDNA3.1, pcDNA3.1-DVL3, shNC or shDVL3 for 72 h. C, D The transfection efficiency were confirmed by western blotting. The expressions of E-cadherin and ZO-1, N-cadherin and Vimentin were detected by western blotting in HCT-8 (E) and SW620 (F) cells transfected with pcDNA3.1 or pcDNA3.1-DVL3 for 72 h, as well as in HCT-8 (G) and SW620 (H) cells transfected with shNC or shDVL3 for 72 h. *P ≤ 0.05

Article Snippet: Primary antibodies against DVL3 and β-catenin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Migration, Transfection, Western Blot

Journal: Journal of translational medicine

Article Title: Disheveled3 enhanced EMT and cancer stem-like cells properties via Wnt/β-catenin/c-Myc/SOX2 pathway in colorectal cancer.

doi: 10.1186/s12967-023-04120-8

Figure Lengend Snippet: Fig. 3 DVL3 was involved in TGF-β-induced EMT. Transwell assay showed the potential migration and invasion of HCT-8 (A) and SW620 (B) cells treated with TGF-β1 for 72 h, or transfected with shNC plus TGF-β1, or shDVL3 plus TGF-β1 for 72 h, as indicated. And then, western blotting was used to analyze the expressions of Snail, E-cadherin, ZO-1, N-cadherin, Vimentin and DVL3 in HCT-8 (C) and SW620 (D) cells transfected with shNC, shNC plus TGF-β1, or shDVL3 plus TGF-β for 72 h, as indicated. *P ≤ 0.05

Article Snippet: Primary antibodies against DVL3 and β-catenin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Transwell Assay, Migration, Transfection, Western Blot

Journal: Journal of translational medicine

Article Title: Disheveled3 enhanced EMT and cancer stem-like cells properties via Wnt/β-catenin/c-Myc/SOX2 pathway in colorectal cancer.

doi: 10.1186/s12967-023-04120-8

Figure Lengend Snippet: Fig. 4 DVL3 regulated CSLCs characteristics and multidrug resistance in CRC cells. A, B The expressions of DVL3, CD44, CD133, SOX2 and c-Myc in tumorspheres and adherent CRC cells (HCT-8 and SW620) were compared by Western blotting. C–F CCK8 assay was used to check the drug sensitivities of tumorspheres and adherent CRC cells treated with VCR or L-OHP for 48 h. The IC50 values of drugs were calculated from cell survival curves using the Bliss method. G Tumorsphere assay was employed to determine the ability of sphere formation in CRC cells transfected with LV-con or LV-DVL3, H and in CRC cells transfected with LV-shNC or LV-shDVL3. I–P CRC cells were transfected with pcDNA3.1, pcDNA3.1-DVL3, shNC or shDVL3 for 24 h, followed by treatment with VCR or L-OHP for additional 48 h. And then, the drug sensitivities of CRC cells were examined by CCK8 assay, and the IC50 values of drugs were calculated from cell survival curves using the Bliss method. Western blotting presented the expressions of CD44, CD133, SOX2 and c-Myc in CRC cells transfected with pcDNA3.1 or pcDNA3.1-DVL3 (Q, R), and shNC or shDVL3 (S, T) for 72 h. *P ≤ 0.05

Article Snippet: Primary antibodies against DVL3 and β-catenin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Western Blot, CCK-8 Assay, Transfection

Journal: Journal of translational medicine

Article Title: Disheveled3 enhanced EMT and cancer stem-like cells properties via Wnt/β-catenin/c-Myc/SOX2 pathway in colorectal cancer.

doi: 10.1186/s12967-023-04120-8

Figure Lengend Snippet: Fig. 5 DVL3 promoted CSLCs and EMT phenotype of CRC via Wnt/β-catenin. A, B The effect of DVL3 on TOPflash and FOPflash luciferase activity were determined by a dual-luciferase reporter assay system in HCT-8 and SW620 cells. C, D The expressions of CD44, CD133, E-cadherin, N-cadherin, DVL3 and β-catenin were detected by western blotting assay in HCT-8 and SW620 cells transfected with shNC, shβ-catenin, or shβ-catenin plus pcDNA3.1-DVL3. E, F Above protein expressions were also determined by western blotting in HCT-8 and SW620 cells treated with shNC, shDVL3, or LiCL plus shDVL3 for 72 h. G, H Transwell assay was used to test the potential migration and invasion of HCT-8 and SW620 cells treated with shNC, shDVL3, or LiCL plus shDVL3 for 72 h. I The ability of tumorsphere formation was examined in CRC cells treated with LiCL, LV-shDVL3, or LiCL plus LV-shDVL3. *P ≤ 0.05

Article Snippet: Primary antibodies against DVL3 and β-catenin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: Luciferase, Activity Assay, Reporter Assay, Western Blot, Transfection, Transwell Assay, Migration

Journal: Journal of translational medicine

Article Title: Disheveled3 enhanced EMT and cancer stem-like cells properties via Wnt/β-catenin/c-Myc/SOX2 pathway in colorectal cancer.

doi: 10.1186/s12967-023-04120-8

Figure Lengend Snippet: Fig. 7 Silencing DVL3 inhibited tumorigenicity and metastasis of CRC cells in vivo. A For tumor xenograft assay in vivo, HCT-8 cells stably expressing LV-shDVL3 or LV-shNC were injected subcutaneously into nude mice. The representative images of tumor xenografts are shown. B Tumor growth curves of tumors in nude mice injected with HCT-8 cells stably expressing LV-shDVL3 or LV-shNC. Once the tumors became palpable, tumor volumes were measured with Vernier calipers every two days. C After the nude mice were killed, tumors xenograft were excised and weighed. D Tumors of LV-shDVL3 group formed at an earlier time point and developed at a faster rate than those of LV-shNC group. E The expression of SOX2, c-Myc, E-cadherin, N-cadherin and DVL3 were determined by western blotting in tumor xenograft with LV-shDVL3 or LV-shNC. F Representative images of lung metastasis and G H&E staining of metastatic tumors in nude mice injected with HCT-8 cells stably expressing LV-shDVL3 or LV-shNC via tail vein. H The statistical data of lung metastasis that developed in the mouse model. *P ≤ 0.05

Article Snippet: Primary antibodies against DVL3 and β-catenin were purchased from Santa Cruz Biotechnology (Santa Cruz, CA, USA).

Techniques: In Vivo, Xenograft Assay, Stable Transfection, Expressing, Injection, Western Blot, Staining