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