hct116  (CLS Cell Lines Service GmbH)

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on Nrf2 activation in DLD-1 and HCT116 cells. ( A ) The level of Nrf2 protein in the cytosolic fraction. ( B ) The level of Nrf2 protein in the nuclear fraction. ( C ) The level of p-Nrf2 protein in the nuclear fraction. Representative Western immunoblots are presented under the graphs ( A – C ). Results (means ± SEM from three separate experiments) are presented as a fold change to control after normalization against the level of actin or lamin. ( D ) The level of Nrf2 binding to DNA. Results are presented as the means ± SEM from three separate experiments percentage of control). Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Activation Assay, Western Blot, Binding Assay

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on the expression of Nrf2 in DLD-1 and HCT116 cells. The values (fold of control) are presented as the means ± SEM from three separate experiments. Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Expressing

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on the expression of selected Nrf2 target genes: SOD and GSTP in DLD-1 and HCT116 cells. ( A ) Levels of SOD and GSTP transcripts. The values (fold of control) are presented as the means ± SEM from three separate experiments. ( B ) Level of SOD and GSTP proteins. Representative Western immunoblots are presented under the graphs. Results (means ± SEM from three separate experiments) are presented as a fold change to control after normalization against the level of actin. Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Expressing, Western Blot

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on the expression of selected Nrf2 target genes: CAT and GPx in DLD-1 and HCT116 cells. ( A ) Levels of CAT and GPx transcripts. The values (fold of control) are presented as the means ± SEM from three separate experiments. ( B ) Levels of CAT and GPx proteins. Representative Western immunoblots are presented under the graphs. Results (means ± SEM from three separate experiments) are presented as a fold change to control after normalization against the level of actin. Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Expressing, Western Blot

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on NF-κB activation in DLD-1 and HCT116 cells. ( A ) The levels of NF-κB p50 and p65 proteins in the cytosolic fraction. ( B ) The levels of NF-κB p50 and p65 proteins in the nuclear fraction. Representative Western immunoblots are presented under the graphs ( A , B ). Results (means ± SEM from three separate experiments) are presented as a fold change to control after normalization against the level of actin or lamin. ( C ) The levels of NF-κB p50 and p65 binding to DNA. Results are presented as the means ± SEM from three separate experiments percentage of control). Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Activation Assay, Western Blot, Binding Assay

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on the expression of NF-κB p50 and p65 in DLD-1 and HCT116 cells. The values (fold of control) are presented as the means ± SEM from three separate experiments. Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Expressing

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on the expression of selected NF-κB target genes: COX-2 and iNOS in DLD-1 and HCT116 cells. ( A ) Levels of COX-2 and iNOS transcripts. The values (fold of control) are presented as the means ± SEM from three separate experiments. ( B ) Levels of COX-2 and iNOS proteins. Representative Western immunoblots are presented under the graphs. Results (means ± SEM from three separate experiments) are presented as a fold change to control after normalization against the level of actin. Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Expressing, Western Blot

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on STAT3 activation in DLD-1 and HCT116 cells. ( A ) The level of STAT3 protein in the cytosolic fraction. ( B ) The level of STAT3 protein in the nuclear fraction. Representative Western immunoblots are presented under the graphs ( A , B ). Results (means ± SEM from three separate experiments) are presented as a fold change to control after normalization against the level of actin or lamin. ( C ) The level of STAT3 binding to DNA. Results are presented as the means ± SEM from three separate experiments percentage of control). Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Activation Assay, Western Blot, Binding Assay

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on the level of p-STAT3 in DLD-1 and HCT116 cells. ( A ) The level of p-STAT3 protein in the nuclear fraction. Representative Western immunoblots are presented under the graphs. Results (means ± SEM from three separate experiments) are presented as a fold change to control after normalization against the level of lamin. ( B ) The ratio of nuclear p-STAT3 and nuclear STAT3 compared with the control group. DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Western Blot

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on the expression of STAT3 in DLD-1 and HCT116 cells. The values (fold of control) are presented as the means ± SEM from three separate experiments. Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Expressing

Journal: Molecules

Article Title: Lichen-Derived Depsides and Depsidones Modulate the Nrf2, NF-κB and STAT3 Signaling Pathways in Colorectal Cancer Cells

doi: 10.3390/molecules26164787

Figure Lengend Snippet: The effect of lichen-derived compounds on the expression of selected STAT3 target gene: Bcl-xl in DLD-1 and HCT116 cells. ( A ) Level of the Bcl-xl transcript. The values (fold of control) are presented as the means ± SEM from three separate experiments. ( B ) Level of the Bcl-xl protein. Representative Western immunoblots are presented under the graphs. Results (means ± SEM from three separate experiments) are presented as a fold change to control after normalization against the level of actin. Asterisks (*) denote statistically significant changes from the control group ( p ≤ 0.05). DMSO , vehicle control; Cap50 , caperatic acid (50 µM); Atra50 , atranorin (50 µM); Leca50 , lecanoric acid (50 µM); Squam50 , squamatic acid (50 µM); Phys25 , physodic acid (25 µM); Salaz50 , salazinic acid (50 µM).

Article Snippet: Human colorectal (HCT116 and DLD-1) cancer cells were obtained from the European Collection of Authenticated Cell Culture (Cell Lines Service, Eppelheim, Germany).

Techniques: Derivative Assay, Expressing, Western Blot

Journal: Marine Drugs

Article Title: Molecular Networking-Guided Isolation of New Etzionin-Type Diketopiperazine Hydroxamates from the Persian Gulf Sponge Cliona celata

doi: 10.3390/md19080439

Figure Lengend Snippet: Bioactivity of compounds 1 and 2 .

Article Snippet: The cultivation of A549 cells (CLS Cell Lines Service, Eppelheim, Germany) was carried out in DMEM:Ham’s F12 medium (1:1) supplemented with 15 mM HEPES, and that of A375 and HCT116 cells (CLS Cell Lines Service, Eppelheim, Germany) was carried out in DMEM supplemented with 4.5 g/L of D-glucose and 110 mg/L of sodium pyruvate.

Techniques: Positive Control

Journal: Oncogene

Article Title: Transglutaminase 2 promotes tumorigenicity of colon cancer cells by inactivation of the tumor suppressor p53

doi: 10.1038/s41388-021-01847-w

Figure Lengend Snippet: A Cell expansion of lentivirally transduced SW480 or (B) HCT-116 cells with two independent shRNAs against TGM2 (shTGM2-1, shTGM2-2) or control (shSCRMBL). C Cell expansion of SW480 and (D) HCT-116 cells after transduction with CRISPR/Cas9 constructs (TGM2 gRNA) against TGM2 or non-target (NT) control. E Mean number of tumorspheres after TGM2 knockdown in SW480 and HCT-116 cells. Data are presented as mean ± SD of at least three independent experiments. F Representative microphotographs of tumorspheres of SW480 cells 14 days after transduction with shTGM2-1, shTGM2-2, or control (shSCRMBL). Shown are fluorescent (tdTOMATO) and brightfield microphotographs. Scale bar, 200 µm. * P < 0.05; ** P < 0.01, Mann–Whitney U test.

Article Snippet: The human colorectal cancer cell lines SW480 and HCT-116 were obtained from CLS Cell Lines Service GmbH (Eppelheim, Germany).

Techniques: Transduction, CRISPR, Construct, MANN-WHITNEY

Journal: Oncogene

Article Title: Transglutaminase 2 promotes tumorigenicity of colon cancer cells by inactivation of the tumor suppressor p53

doi: 10.1038/s41388-021-01847-w

Figure Lengend Snippet: A Time-lapse imaging of SW480 cells transduced with shTGM2-1, shTGM2-2 or shSCRMBL. Shown are cumulative cell death events over time determined by single cell tracking. P value was calculated by log-rank test. B Percentage of apoptotic SW480 cells determined by Annexin V/7-AAD staining 72 hours after TGM2 knockdown. C Percentage of Caspase-3 positive SW480 cells 72 hours after TGM2 knockdown. D – F All experiments were repeated in HCT-116 cells transduced with shTGM2-1, shTGM2-2 or shSCRMBL. D Time lapse imaging showing the cumulative cell death events. E Percentage of Annexin V positive and ( F) Caspase-3 positive HCT-116 cells 72 hours after TGM2 knockdown. Results are presented as mean ± SD of three independent experiments. *** P < 0.001, Mann–Whitney U test.

Article Snippet: The human colorectal cancer cell lines SW480 and HCT-116 were obtained from CLS Cell Lines Service GmbH (Eppelheim, Germany).

Techniques: Imaging, Transduction, Single Cell Tracking, Staining, MANN-WHITNEY

Journal: Oncogene

Article Title: Transglutaminase 2 promotes tumorigenicity of colon cancer cells by inactivation of the tumor suppressor p53

doi: 10.1038/s41388-021-01847-w

Figure Lengend Snippet: A – C Gene expression profiling by RNA-seq of SW480 cells after transduction with either shTGM2-1 or shSCRMBL. A Unsupervised hierarchical clustering of the top 1000 differentially expressed genes (DEGs) upon TGM2 knockdown across the four biological replicates. B MA plot relating p values for all differentially expressed genes between shTGM2-1 and shSCRMBL from four biological replicates. Red dots indicate significantly regulated genes (adjusted P < 0.05). List of regulated genes is presented in Supplementary Table S . C Scatter plot of gene set enrichment analysis of DEGs relating the Q-value for Hallmark gene-set signatures. The top 16 enriched pathways are shown ( P < 0.05, Fold change ≥2). The color and size of each dot represent the Rich factor and the number of DEGs mapped to the indicated pathway, respectively. D Proteome analysis of regulated proteins involved in apoptosis upon shRNA-mediated TGM2 knockdown. Representative blot of Proteome Profiler Array™-Human Apoptosis Array analysis of SW480 cells. The regulation of protein expression of phosphorylated p53 variants is shown. E – H Quantification of p53 and phosphorylated p53 (S15, S46, and S392) upon TGM2 knockdown in SW480 ( E , G ) and HCT-116 ( F , H ) cells via Simple Western technology ( n = 3; Mann–Whitney U test).

Article Snippet: The human colorectal cancer cell lines SW480 and HCT-116 were obtained from CLS Cell Lines Service GmbH (Eppelheim, Germany).

Techniques: Expressing, RNA Sequencing Assay, Transduction, shRNA, Western Blot, MANN-WHITNEY

Journal: Oncogene

Article Title: Transglutaminase 2 promotes tumorigenicity of colon cancer cells by inactivation of the tumor suppressor p53

doi: 10.1038/s41388-021-01847-w

Figure Lengend Snippet: A Representative images of proximity ligation assay (PLA) of TGM2 and p53 in SW480 cells. Cells incubated only with TGM2 antibody served as negative control (I). Protein–protein interaction of TGM2 and p53(S15) was visualized using hybridization probes labeled with Texas Red (II). Nuclei were stained with DAPI (blue). B Quantification of TGM2-p53 interaction and associated technical controls (Ctrl). Technical controls demonstrate the specificity of PLA signals. Each dot represents one cell. Mean value of PLA dots per cell is shown by the black line. C Representative images of proximity ligation assay of TGM2 and p53 in patient-derived normal epithelial cells (I) and corresponding colon cancer cells (II). D Quantification of TGM2-p53 interaction in primary patient material. (Significance was calculated using Kruskal–Wallis test). E Co-immunoprecipitation (Co-IP) of endogenous TGM2 and p53 or phosphorylated p53(S15) in SW480, HCT-116 p53 wildtype cells (wt) and HCT-116 p53 knockout cells (−/−). F Super-resolved image of a HCT-116 cell immunostained for TGM2 (red) and p53(S15) (cyan). A zoom-in of the highlighted region is shown on the right. White regions indicate overlapping signal of TGM2 and p53(S15) (yellow arrowheads). Scale bars represent 5 µm and 1 µm, respectively.

Article Snippet: The human colorectal cancer cell lines SW480 and HCT-116 were obtained from CLS Cell Lines Service GmbH (Eppelheim, Germany).

Techniques: Proximity Ligation Assay, Incubation, Negative Control, Hybridization, Labeling, Staining, Derivative Assay, Immunoprecipitation, Co-Immunoprecipitation Assay, Knock-Out

Journal: Oncogene

Article Title: Transglutaminase 2 promotes tumorigenicity of colon cancer cells by inactivation of the tumor suppressor p53

doi: 10.1038/s41388-021-01847-w

Figure Lengend Snippet: A – C HCT-116 p53 wildtype cells (wt) and HCT-116 p53 knockout cells (−/−) were transduced with either shTGM2-1, shTGM2-2, or shSCRMBL. Time-lapse imaging and proliferation assay were performed to determine a rescue from cell death upon TGM2 knockdown. A Fold change of cell number of HCT-116 p53 wt and HCT-116 p53 −/− cells upon TGM2 knockdown in comparison to shSCRMBL control determined at day three after transduction. Data are presented as mean ± SD of three independent experiments (** P < 0.01, Mann–Whitney U test). B Single cell tracking of HCT-116 p53 wt and HCT-116 p53 − /− cells after TGM2 knockdown with shTGM2-1 and (C) shTGM2-2. Cumulative cell death events are shown over time (*** P < 0.001, Log-rank test). D Direct visualization of p53 activation upon TGM2 knockdown by time-lapse video-microscopy. Sequence of phase contrast images, tdTOMATO fluorescence of shTGM2-1 and p53-driven destabilized GFP reporter , depicting the same field of view over the time course of 30 hours as indicated in the corresponding panels in I–VIII. The yellow circles designate tracked cells over time. (I–VIII) show corresponding sequence of fluorescence images taken at the same time points as the phase contrast images. (I) Shown are two representative HCT-116 cells. (II and III) 6-8 hours after lentiviral transduction of shTGM2-1 both HCT-116 cells express the red fluorescent tdTOMATO reporter, indicating a knockdown of TGM2. (IV-VI) Another 4–10 hours later both cells express the green fluorescent (GFP) p53 reporter, indicating the induction of p53 activity. (VII and VIII) About 24 hours after transduction both HCT-116 cells subsequently undergo apoptosis (white arrows). Movie S shows all assembled images (3 min temporal resolution) of the same sequence.

Article Snippet: The human colorectal cancer cell lines SW480 and HCT-116 were obtained from CLS Cell Lines Service GmbH (Eppelheim, Germany).

Techniques: Knock-Out, Transduction, Imaging, Proliferation Assay, MANN-WHITNEY, Single Cell Tracking, Activation Assay, Microscopy, Sequencing, Fluorescence, Activity Assay