Journal: Oncology Letters

Article Title: Differential effect of psoralidin in enhancing apoptosis of colon cancer cells via nuclear factor-κB and B-cell lymphoma-2/B-cell lymphoma-2-associated X protein signaling pathways

doi: 10.3892/ol.2015.3861

Figure Lengend Snippet: Psoralidin treatment reduces the viability of SW480 cells. Values are expressed as the mean ± standard deviation. *P<0.05 and **P<0.01 compared with the control group.

Article Snippet: Cell line and culture The SW480 human colon cancer cell line was obtained from the Department of Oncology (Central Hospital of Jingzhou, Jingzhou, China).

Techniques: Standard Deviation, Control

Journal: Oncology Letters

Article Title: Differential effect of psoralidin in enhancing apoptosis of colon cancer cells via nuclear factor-κB and B-cell lymphoma-2/B-cell lymphoma-2-associated X protein signaling pathways

doi: 10.3892/ol.2015.3861

Figure Lengend Snippet: Psoralidin enhances cellular apoptosis of SW480 cells. Effect of psoralidin on the cellular apoptosis of SW480 cells examined by (A) flow cytometry and (B) DAPI staining assay (magnification, ×200). Values are expressed as the mean ± standard deviation. *P<0.05 and **P<0.01 compared with control group. FCM, flow cytometry.

Article Snippet: Cell line and culture The SW480 human colon cancer cell line was obtained from the Department of Oncology (Central Hospital of Jingzhou, Jingzhou, China).

Techniques: Flow Cytometry, Staining, Standard Deviation, Control

Journal: Oncology Letters

Article Title: Differential effect of psoralidin in enhancing apoptosis of colon cancer cells via nuclear factor-κB and B-cell lymphoma-2/B-cell lymphoma-2-associated X protein signaling pathways

doi: 10.3892/ol.2015.3861

Figure Lengend Snippet: Psoralidin treatment enhances caspase-3 activity of SW480 cells. Values are expressed as the mean ± standard deviation. *P<0.05 and **P<0.01 compared with control group.

Article Snippet: Cell line and culture The SW480 human colon cancer cell line was obtained from the Department of Oncology (Central Hospital of Jingzhou, Jingzhou, China).

Techniques: Activity Assay, Standard Deviation, Control

Journal: Oncology Letters

Article Title: Differential effect of psoralidin in enhancing apoptosis of colon cancer cells via nuclear factor-κB and B-cell lymphoma-2/B-cell lymphoma-2-associated X protein signaling pathways

doi: 10.3892/ol.2015.3861

Figure Lengend Snippet: Psoralidin reduces the NF-κB activity of SW480 cells. Values are expressed as the mean ± standard deviation. *P<0.05 and **P<0.01 compared with control group. NF-κB, nuclear factor-κB; OD, optical density.

Article Snippet: Cell line and culture The SW480 human colon cancer cell line was obtained from the Department of Oncology (Central Hospital of Jingzhou, Jingzhou, China).

Techniques: Activity Assay, Standard Deviation, Control

Journal: Oncology Letters

Article Title: Differential effect of psoralidin in enhancing apoptosis of colon cancer cells via nuclear factor-κB and B-cell lymphoma-2/B-cell lymphoma-2-associated X protein signaling pathways

doi: 10.3892/ol.2015.3861

Figure Lengend Snippet: Psoralidin inhibits Bcl-2 and enhances Bax protein expression in SW480 cells. (A and B) Effect of psoralidin on Bcl-2 protein expression of SW480 cells by western blotting assay and statistical analysis of Bcl-2 protein expression of SW480 cells. (C and D) Effects of psoralidin on Bax protein expression of SW480 cells by western blotting assay and statistical analysis of Bax proteins expression of SW480 cells. Values are expressed as the mean ± standard deviation. *P<0.05 and **P<0.01 compared with control group. Bcl-2, B-cell lymphoma-2; Bax, Bcl-2-associated X protein.

Article Snippet: Cell line and culture The SW480 human colon cancer cell line was obtained from the Department of Oncology (Central Hospital of Jingzhou, Jingzhou, China).

Techniques: Expressing, Western Blot, Standard Deviation, Control

Journal: Oncology Letters

Article Title: Differential effect of psoralidin in enhancing apoptosis of colon cancer cells via nuclear factor-κB and B-cell lymphoma-2/B-cell lymphoma-2-associated X protein signaling pathways

doi: 10.3892/ol.2015.3861

Figure Lengend Snippet: Bcl-2 inhibitor enhances the effect of psoralidin on SW480 cells. (A) Effect of Bcl-2 inhibitor on Bcl-2 protein expression of SW480 cells by western blotting assays; (B) statistical analysis of Bcl-2 protein expression of SW480 cells; and (C) Bcl-2 inhibitor enhances the effect of psoralidin on viability of SW480 cells. Values are expressed as the mean ± standard deviation. **P<0.01 compared with control group; ##P<0.01 compared with PSO (10) group. Bcl-2, B-cell lymphoma-2; PSO, psoralidin; ABT, ABT-737.

Article Snippet: Cell line and culture The SW480 human colon cancer cell line was obtained from the Department of Oncology (Central Hospital of Jingzhou, Jingzhou, China).

Techniques: Expressing, Western Blot, Standard Deviation, Control

Journal: Heliyon

Article Title: A novel lnc-LAMC2-1:1 SNP promotes colon adenocarcinoma progression by targeting miR-216a-3p/HMGB3

doi: 10.1016/j.heliyon.2022.e12342

Figure Lengend Snippet: The lnc-LAMC2-1:1 SNP is a sponge of miR-216a-3p . Bar graphs of (a) and (b) show the relative luciferase activity of vectors containing SW480. A dual-luciferase reporter assay was performed, and the co-transfection of lnc-LAMC2-1:1 SNP and miR-216a-3p reduced the luciferase activity.

Article Snippet: The SW480 cell line was obtained from COSMOBIO Company in China and was grown in DMEM supplemented with 10% FBS (GIBCO) and 1% antibiotics (50 U/mL penicillin and 50 μg/mL streptomycin) in a humidified atmosphere of 5% CO2 at 37 °C.

Techniques: Luciferase, Activity Assay, Reporter Assay, Cotransfection

Journal: Heliyon

Article Title: A novel lnc-LAMC2-1:1 SNP promotes colon adenocarcinoma progression by targeting miR-216a-3p/HMGB3

doi: 10.1016/j.heliyon.2022.e12342

Figure Lengend Snippet: The lnc-LAMC2-1:1 SNP positively regulates HMGB3 by sponging miR-126-3p (a) mRNA expression of HMGB3 in SW480 cells. Cells were transfected with mimics of NC, miR-126-3p mimics, ASO-NC, and ASO-miR-126-3p. (b) mRNA expression of HMGB3 in SW480 cells. Cells were transfected with pcDNA3.1, lnc-LAMC2-1:1 SNP , and lnc-LAMC2-1:1-wt. (c) Complementary sequences of HMGB3 and miR-126-3p in the StarBase database. (d) A dual-luciferase reporter assay was performed, and the co-transfection of miR-126-3p and HMGB3 reduced the luciferase activity.

Article Snippet: The SW480 cell line was obtained from COSMOBIO Company in China and was grown in DMEM supplemented with 10% FBS (GIBCO) and 1% antibiotics (50 U/mL penicillin and 50 μg/mL streptomycin) in a humidified atmosphere of 5% CO2 at 37 °C.

Techniques: Expressing, Transfection, Luciferase, Reporter Assay, Cotransfection, Activity Assay

Journal: iScience

Article Title: APC-driven actin nucleation powers collective cell dynamics in colorectal cancer cells

doi: 10.1016/j.isci.2023.106583

Figure Lengend Snippet: APC-m4 decreases the levels of F-actin and key molecular components at cell junctions All data are from SW480 stably expressing APC-WT or APC-m4. (A) Schematic showing the position of APC domains (colored boxes, from left to right): Dark blue boxes represent oligomerization domains, green boxes are the armadillo repeats, yellow boxes are 15 and 20 amino acid repeats, brown boxes are serine-alanine-methionine-proline (SAMP) motifs, dark green boxes are nuclear localization signal motifs (NLS), light blue box shows the basic domain (that contains one oligomerization domain shown in blue). Above the cartoon, some APC partners to the corresponding interaction sites have been stated. Red dots indicate the location of the APC-m4 mutation (L2539A I2541A) in the oligomerization motif within the basic domain. The lines below the schematic represent the two cell lines used in this study, and the corresponding length of the APC protein that they express. These are SW480 cell line, which expresses a truncated product lacking both signaling and cytoskeletal functions (1–1388 amino acid), and HCT116 cell line that expresses full-length APC (1–2843 amino acid). (B) Qualitative graph showing the percentage of cells that belong to the “normal”, “discontinuous”, and “round” category assigned in function of the appearance of the F-actin label at the cell junctions (see ). two-way ANOVA Sidak’s multiple comparisons test was performed to find the statistical differences. “∗∗∗∗” is p < 0.0001, “∗∗∗” is p < 0.001, and “ns” is not significant. N = 3 independent repeats for each condition; n(APC-WT) = 137 cells in total, n(APC-m4) = 95 cells in total. (C) Representative immunofluorescence images of F-actin and occludin. Scale bar = 20 μm. (D) Violin plot showing the average F-actin intensity at the cell junctions. N = 4 independent repeats for each condition; n(APC-WT) = 107 junctions in total, n(APC-m4) = 241 junctions in total. (E) Violin plot showing the average occludin intensity at the cell junctions. N = 4 independent replicates for each condition; n(APC-WT) = 107 junctions in total, n(APC-m4) = 195 junctions in total. Data in D and E are displayed as “superplots” showing the mean of the different replicates (circles) and the distribution of “n junctions analyzed” (color-coded dots) was superimposed as violin plot. Solid line is the mean and paired two-tailed t test was used to find the statistical differences using N = 4 replicates. “∗” is p < 0.05. (F) Representative scanning electron microscopy images of the cell junctions. Scale bar = 1 μm, inset = 0.5 μm. See also Figure S1 .

Article Snippet: SW480 human colorectal cell line , Merck , RRID:CVCL_0546.

Techniques: Stable Transfection, Expressing, Mutagenesis, Immunofluorescence, Two Tailed Test, Electron Microscopy

Journal: iScience

Article Title: APC-driven actin nucleation powers collective cell dynamics in colorectal cancer cells

doi: 10.1016/j.isci.2023.106583

Figure Lengend Snippet: APC-m4 perturbs cell junction dynamics of epithelial monolayers All data are from SW480 stably expressing APC-WT or APC-m4 and transiently transfected with E-cadherin-GFP. (A) Representative time lapse images showing E-cadherin-GFP at cell junctions during a 20-min observation window. Scale bar = 5 μm. The color gradient shows the varying signal intensity. White arrows show disrupted cell junction vertexes. (B) Cartoon representing junction length considered for analysis. Next, violin plot showing fold change in cell junction length during the 20 min observation window, taking as a reference the time 0 min. N = 3 independent repeats for each condition; junctions: n = 9 per condition for the 11 planes of the time lapse, being that n = 99 junctions total per condition. (C) Cartoon representing junction angle considered for analysis. Next, a graph showing the percentage of cell junctions with junction angles between 0° and 20°. Cell junction angle at time 0 = 0°. N = 3 independent repeats for each condition; n = 99 junctions total per condition. (D) Cartoon representing junction velocity considered for analysis. Next, representative kymographs of cell junctions and violin plot showing cell junction velocity. N = 3 independent repeats for each condition; n(APC-WT) = 43 junctions, n(APC-m4) = 53 junctions. (E) Cartoon representing criteria of normal and disrupted junctions considered for analysis. Next, violin plot showing cells with no (0), one (1) or both (2) junction vertexes disrupted. N = 3 independent repeats for each condition; n(APC-WT) = 506 vertexes in total, n(APC-m4) = 495 vertexes in total. (F) Cartoon representing junction velocity considered for analysis. Next, representative kymographs of vertexes of cell junctions and violin plot showing vertex velocity. N = 3 independent repeats for each condition; n(APC-WT) = 88 vertexes in total, n(APC-m4) = 104 vertexes in total. (G) Cartoon representing how the difference between the velocities of two vertexes at the same cell junction was analyzed and violin plot showing those differences in vertex velocities within a junction. N = 3 independent repeats for each condition; n(APC-WT) = 44 junctions in total, n(APC-m4) = 52 junctions in total. Data in B and D-F are displayed as “superplots” showing the mean of the different replicates (circles) and the distribution of “n” junctions or vertexes analyzed, as stated in each graph, (color-coded dots) was superimposed as violin plot. Black lines are the mean and standard deviation; paired two-tailed t test was performed to find the statistical differences, except for B that ratio paired two-tailed t test was performed, using N = 3 independent replicates for all graphs. “∗” is p < 0.05, “ns” is not significant. See also .

Article Snippet: SW480 human colorectal cell line , Merck , RRID:CVCL_0546.

Techniques: Stable Transfection, Expressing, Transfection, Standard Deviation, Two Tailed Test

Journal: iScience

Article Title: APC-driven actin nucleation powers collective cell dynamics in colorectal cancer cells

doi: 10.1016/j.isci.2023.106583

Figure Lengend Snippet: Expression of APC-m4 mutant results in enlarged cells (A) Experiment workflow. (B) Representative segmentation images from cellpose of HCT116 cells stably expressing APC-WT and APC-m4. (C) Violin plot showing the size of individual HCT116 cells shown in (B). n(APC-WT) = 1487 cells in total, n(APC-m4) = 1586 cells in total. (D) Representative segmentation images from cellpose of SW480 cells stably expressing APC-WT and APC-m4. (E) Violin plot showing the size of individual cells SW480 shown in (D). n(APC-WT) = 157 cells in total, n(APC-m4) = 98 cells in total. For panels (C) and (E), The solid line is median and dotted lines are quartiles. Mann-Whitney U test was performed to find the statistical difference. “∗∗∗∗” is p < 0.0001. (F) Violin plots showing the size of individual SW480 cells forming monolayers treated with DMSO or SMIFH2. The solid line is the median and dotted lines are quartiles. One-way ANOVA Sidak’s test was performed to find statistical differences. “∗∗∗∗” is p < 0.0001, and “ns” is not significant. n(APC-WT DMSO) = 208, n(APC-m4 DMSO) = 165, n(APC-WT SMIFH2) = 187, and n(APC-m4 SMIFH2) = 147. Data from C, E, and F graphs are from three independent repeats. (G) Representative graphs showing FACS data of HCT116 APC-WT and APC-m4 stably expressing cells. (H) From the left to the right: Violin plots showing the percentage of HCT116 APC-WT and APC-m4 cells in G1, S, and G2. N = 3 independent experimental repeats. The solid line is the median and dotted lines are quartiles. Mann-Whitney U test was performed to find the statistical difference. “ns” is not significant.

Article Snippet: SW480 human colorectal cell line , Merck , RRID:CVCL_0546.

Techniques: Expressing, Mutagenesis, Stable Transfection, MANN-WHITNEY

Journal: iScience

Article Title: APC-driven actin nucleation powers collective cell dynamics in colorectal cancer cells

doi: 10.1016/j.isci.2023.106583

Figure Lengend Snippet:

Article Snippet: SW480 human colorectal cell line , Merck , RRID:CVCL_0546.

Techniques: Recombinant, Modification, Stable Transfection, Expressing, Plasmid Preparation, Software, Microscopy, Chemotaxis Assay, Migration