colo 205 ccl 222 colorectal cancer cells  (ATCC)

Journal: Frontiers in Oncology

Article Title: Inhibition of colorectal cancer progression through conformation-specific targeting of ADAM10 metalloprotease

doi: 10.3389/fonc.2025.1704436

Figure Lengend Snippet: 1H5 reduces viability, proliferation, and colony formation in colorectal cancer (CRC) cells. (A) Cell viability of COLO205, SW620, and DLD-1 cells measured by Alamar Blue after 48 h and 72 h of treatment with IgG control (40 µg/mL) or 1H5 (5–40 µg/mL). Viability values were normalized to the IgG control for visualization. For statistical testing across cell lines, normalized values were converted to Δ-viability (treatment – 100), and each time point (48 h and 72 h) was analyzed separately using two-way ANOVA (cell line × concentration) followed by Dunnett’s multiple-comparison test. (B) Real-time proliferation measured by IncuCyte. Phase-area confluence was tracked every 4 h and normalized to baseline. Statistical comparisons were performed using two-way ANOVA with Dunnett’s correction. (C) Representative images of crystal violet–stained colonies after 10 days of treatment. (D) Quantification of colony numbers from panel (C) . Group differences were analyzed using the Kruskal–Wallis test with multiple-comparison correction. Data represent mean ± SD from biological replicates. Significance levels: *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001 .

Article Snippet: Human colorectal cancer cell lines COLO205, DLD-1, and SW620 (ATCC, USA) were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: Control, Concentration Assay, Comparison, Staining

Journal: Frontiers in Oncology

Article Title: Inhibition of colorectal cancer progression through conformation-specific targeting of ADAM10 metalloprotease

doi: 10.3389/fonc.2025.1704436

Figure Lengend Snippet: 1H5 specifically targets ADAM10. (A) Proliferation curves of shRenilla and shADAM10 knockdown cells (COLO205, SW620, and DLD-1) measured by live-cell imaging over 96 h following doxycycline induction. Cells were seeded in the presence of 1 µg/mL doxycycline to induce shRNA expression. Phase area was normalized to baseline (0 h). (B) Relative proliferation of doxycycline-induced shRenilla and shADAM10 cells following treatment with 40 µg/mL 1H5. 1H5 was added 24 h after seeding, and proliferation was monitored by live-cell imaging over time. Data represent three biological replicates and are shown as mean ± SD. ****p < 0.0001, ***p < 0.001, **p < 0.01 by two-way ANOVA.

Article Snippet: Human colorectal cancer cell lines COLO205, DLD-1, and SW620 (ATCC, USA) were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: Knockdown, Live Cell Imaging, shRNA, Expressing

Journal: Frontiers in Oncology

Article Title: Inhibition of colorectal cancer progression through conformation-specific targeting of ADAM10 metalloprotease

doi: 10.3389/fonc.2025.1704436

Figure Lengend Snippet: 1H5 inhibits Notch signaling in CRC cell lines. (A) Basal protein expression of NICD, NOTCH1, phosphorylated EGFR (p-EGFR), and total EGFR in DLD-1, SW620, and COLO205 cells. GAPDH served as a loading control. Western blots were performed using two biological replicates. (B) Western blot analysis of NICD and NOTCH1 in COLO205 cells treated with 1H5 (5–40 µg/mL), γ-secretase inhibitor (GSI; 20 or 100 µM), IgG control (100 µg/mL), or DMSO (100 µM) for 48 (h) GAPDH was used as a loading control. Two biological replicates were performed. (C) HES1 mRNA expression in COLO205 cells treated with IgG control (20 µg/mL) or 1H5 (5 or 20 µg/mL) for 48 h Data are presented as mean ± SD. Statistical significance was assessed using the Kruskal–Wallis test followed by Dunn’s multiple-comparison test; p < 0.05. (D) Western blot analysis of NICD and NOTCH1 in SW620 cells treated with 1H5 (5 or 20 µg/mL), GSI (20 µM), IgG control (20 µg/mL), or DMSO (100 µM) for 48 (h) GAPDH was used as a loading control. Two biological replicates were performed. (E) HES1 mRNA expression in SW620 cells treated with IgG control (20 µg/mL) or 1H5 (5 or 20 µg/mL) for 48 (h) Data are presented as mean ± SD. Statistical significance was assessed using the Kruskal–Wallis test with Dunn’s post hoc test; *p < 0.05 .

Article Snippet: Human colorectal cancer cell lines COLO205, DLD-1, and SW620 (ATCC, USA) were maintained in RPMI-1640 medium supplemented with 10% fetal bovine serum (FBS) and 1% penicillin/streptomycin (P/S).

Techniques: Expressing, Control, Western Blot, Comparison

Journal: Communications Biology

Article Title: Engineering a single-chain immunoglobulin scaffold loaded with a latent-releasable cytotoxic pore-forming peptide

doi: 10.1038/s42003-025-09066-9

Figure Lengend Snippet: a Sequences of modified peptides, using M70 as a reference for non-cytotoxic/producible peptides and melittin as a reference for cytotoxic/non-producible peptides. Highlighted in red are melittin residues, in blue M70 residues, and in bold black newly introduced residues. b Amphipathic properties of the peptides. Helical wheel representations rendered with the Heliquest server . c Physicochemical parameters of the full sequences, including mean hydrophobicity (); hydrophobic moment (<μH>) and charge, estimated using the same server . d Representative titration experiment of COLO205 cell membrane permeabilization to 7-ADD after incubation with melittin, the modified peptides, or M70. One representative out of at least three independent replicates with similar results is shown. Mean values of two technical replicates are represented in the plot. Bar chart: EC 50 values determined from three independent experiments, with bars showing mean values. e Expression levels of the scAtezo-Pmod2 hybrid in HEK 293F cells with the peptide integrated at different positions: within the HC-LC linker, at the C-terminus of the LC, or at the C-terminus of the HC. Created with BioRender.com.

Article Snippet: The Colorectal cancer cell line COLO205 and the lung cancer cell lines NCI-H2122 and NCI-H2228 (all purchased from ATCC, USA) were cultured in RPMI 1640 medium (Merck) supplemented with 10% fetal bovine serum.

Techniques: Modification, Titration, Membrane, Incubation, Expressing

Journal: Communications Biology

Article Title: Engineering a single-chain immunoglobulin scaffold loaded with a latent-releasable cytotoxic pore-forming peptide

doi: 10.1038/s42003-025-09066-9

Figure Lengend Snippet: a Diagram showing residues added to Pmod2 following matriptase cleavage, along with sequences of Pmod2-based peptides. b COLO205 cell membrane permeabilization assays comparing the potency of Pmod2-1 (cyan) and Pmod2-2 (gray) with respect to Pmod2 (orange). One representative out of at least three independent replicates with similar results is shown. Mean values of two technical replicates are represented in the plot. Bar chart: EC 50 values ± SD determined from three independent experiments, with bars showing mean values. c CD spectra in HFIP and d helical wheel representations of Pmod2-1 and Pmod2-2. e SEC profile of purified scAtezo-Pmod2-2 and binding kinetics to PD-L1 analyzed by BLI. Red lines represent experimental data, and black lines show the global fit.

Article Snippet: The Colorectal cancer cell line COLO205 and the lung cancer cell lines NCI-H2122 and NCI-H2228 (all purchased from ATCC, USA) were cultured in RPMI 1640 medium (Merck) supplemented with 10% fetal bovine serum.

Techniques: Membrane, Circular Dichroism, Purification, Binding Assay

Journal: International Journal of Molecular Sciences

Article Title: Antitumor Activity by an Anti-CD44 Variant 9 Monoclonal Antibody in Gastric and Colorectal Cancer Xenograft Models

doi: 10.3390/ijms26189170

Figure Lengend Snippet: Flow cytometry analysis of C 44 Mab-1-mG 2a to CD44v9-positive cancer cell lines. ( A ) MKN45 and COLO205 were treated with 0.01, 0.1, and 1 µg/mL of C 44 Mab-1-mG 2a . ( B ) MKN45 and COLO205 were treated with 0.01, 0.1, and 1 µg/mL of control mIgG 2a . Then, the cells were treated with Alexa Fluor 488-conjugated anti-mouse IgG. Fluorescence data were analyzed using the SA3800 Cell Analyzer.

Article Snippet: CHO-K1 and COLO205 (colorectal cancer) cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Flow Cytometry, Control, Fluorescence

Journal: International Journal of Molecular Sciences

Article Title: Antitumor Activity by an Anti-CD44 Variant 9 Monoclonal Antibody in Gastric and Colorectal Cancer Xenograft Models

doi: 10.3390/ijms26189170

Figure Lengend Snippet: ADCC by C 44 Mab-1-mG 2a against CHO/CD44v3–10, MKN45, and COLO205. ADCC induced by C 44 Mab-1-mG 2a or control mouse IgG 2a (mIgG 2a ) against CHO/CD44v3–10 ( A ), MKN45 ( B ), and COLO205 ( C ). Calcein AM-labeled target cells (CHO/CD44v3–10, MKN45, and COLO205) were incubated with the effector splenocytes in the presence of 100 μg/mL of C 44 Mab-1-mG 2a or control mIgG 2a . Following a 4.5 h incubation, the Calcein release into the medium was measured. Values are shown as mean ± SEM. Asterisks indicate statistical significance (* p < 0.05; Two-tailed unpaired t -test).

Article Snippet: CHO-K1 and COLO205 (colorectal cancer) cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Control, Labeling, Incubation, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Antitumor Activity by an Anti-CD44 Variant 9 Monoclonal Antibody in Gastric and Colorectal Cancer Xenograft Models

doi: 10.3390/ijms26189170

Figure Lengend Snippet: CDC by C 44 Mab-1-mG 2a against CHO/CD44v3–10, MKN45, and COLO205. CDC induced by C 44 Mab-1-mG 2a or control mouse IgG 2a (mIgG 2a ) against CHO/CD44v3–10 ( A ), MKN45 ( B ), and COLO205 ( C ). Calcein AM-labeled target cells (CHO/CD44v3–10, MKN45, and COLO205) were incubated with complements and C 44 Mab-1-mG 2a or control mIgG 2a . Following incubation for 4.5 h at 37 °C, the Calcein release into the medium was measured. Values are shown as mean ± SEM. Asterisks indicate statistical significance (* p < 0.05; Two-tailed unpaired t -test).

Article Snippet: CHO-K1 and COLO205 (colorectal cancer) cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Control, Labeling, Incubation, Two Tailed Test

Journal: International Journal of Molecular Sciences

Article Title: Antitumor Activity by an Anti-CD44 Variant 9 Monoclonal Antibody in Gastric and Colorectal Cancer Xenograft Models

doi: 10.3390/ijms26189170

Figure Lengend Snippet: Antitumor activity of C 44 Mab-1-mG 2a against CHO/CD44v3–10, MKN45, and COLO205 xenograft. ( A – C ) CHO/CD44v3–10 ( A ), MKN45 ( B ), and COLO205 ( C ) were subcutaneously injected into BALB/c nude mice (day 0). An amount of 100 μg of C 44 Mab-1-mG 2a or control mouse IgG 2a (mIgG 2a ) was intraperitoneally injected into each mouse on day 8. Additional antibodies were injected on day 14 (MKN45 and COLO205) or 14 and 21 (CHO/CD44v3–10) (red arrows). The tumor volume is represented as the mean ± SEM. ** p < 0.01, * p < 0.05 (ANOVA with Sidak’s multiple comparisons test). ( D – F ) The mice treated with the mAbs were euthanized on day 21 (MKN45 and COLO205) or 28 (CHO/CD44v3–10). The xenograft weights were measured. Values are presented as the mean ± SEM. ** p < 0.01 (Two-tailed unpaired t -test). ( G – I ) Body weights of xenograft-bearing mice treated with the mAbs. There is no statistical difference.

Article Snippet: CHO-K1 and COLO205 (colorectal cancer) cell lines were obtained from the American Type Culture Collection (Manassas, VA, USA).

Techniques: Activity Assay, Injection, Control, Two Tailed Test