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ML329 decreases cell viability; reduces the expression of MITF, KIT, BCL2, and CDK2; and impairs the cell cycle in GIST cell lines GIST cells: GIST-T1 (A) and GIST 430/654 (G) were incubated with various concentrations of ML329 on different days. Cell viability was measured (∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; two-way ANOVA analysis of significance, Tukey’s multiple comparisons test). Sigmoidal 4PL representations of cytotoxicity for GIST-T1 after 3 days of ML329 incubation (5 μM) (B) and for GIST 430/654 after 10 days of ML329 incubation (5 μM) were performed (H). MITF activity was measured using the TRPM-1-luciferase gene reporter in GIST-T1 after 1 day of ML329 incubation (5 μM) (C) and in GIST 430/654 after 8 days of ML329 incubation (5 μM) (I) (∗∗ p < 0.01; unpaired t test). Caspase 3/7 activity was measured in GIST-T1 (D) and GIST 430/657(J), incubated with 10 μM ML329 after 24 h of administration (∗∗ p < 0.01, ∗∗∗ p < 0.001; unpaired t test). The relative number of cells (%) was plotted, showing cell-cycle phases in GIST-T1 treated with ML329 (5 μM) at days 5 and 7 (E) and GIST 430/654 treated with ML329 (10 μM) at days 7 and 10 (K). Results were analyzed using the Dean/Jett/Fox model and FlowJo 7.0 software. (∗ p < 0.05, ∗∗ p < 0.01; unpaired t test.) Western blots for KIT, MITF, BCL2, and CDK2 are shown after treatment with ML329 in GIST-T1 after 3 days and GIST 430/654 after 10 days (F and L, respectively). β-actin was used as a loading control. Ratios of proteins blotted versus β-actin are shown. All experiments were performed at least three times, and the blots represent the results of several experiments.

Journal: Molecular Therapy Oncology

Article Title: Preclinical study of microphthalmia-associated transcription factor inhibitor ML329 in gastrointestinal stromal tumor growth

doi: 10.1016/j.omton.2025.200983

Figure Lengend Snippet: ML329 decreases cell viability; reduces the expression of MITF, KIT, BCL2, and CDK2; and impairs the cell cycle in GIST cell lines GIST cells: GIST-T1 (A) and GIST 430/654 (G) were incubated with various concentrations of ML329 on different days. Cell viability was measured (∗∗∗ p < 0.001, ∗∗∗∗ p < 0.0001; two-way ANOVA analysis of significance, Tukey’s multiple comparisons test). Sigmoidal 4PL representations of cytotoxicity for GIST-T1 after 3 days of ML329 incubation (5 μM) (B) and for GIST 430/654 after 10 days of ML329 incubation (5 μM) were performed (H). MITF activity was measured using the TRPM-1-luciferase gene reporter in GIST-T1 after 1 day of ML329 incubation (5 μM) (C) and in GIST 430/654 after 8 days of ML329 incubation (5 μM) (I) (∗∗ p < 0.01; unpaired t test). Caspase 3/7 activity was measured in GIST-T1 (D) and GIST 430/657(J), incubated with 10 μM ML329 after 24 h of administration (∗∗ p < 0.01, ∗∗∗ p < 0.001; unpaired t test). The relative number of cells (%) was plotted, showing cell-cycle phases in GIST-T1 treated with ML329 (5 μM) at days 5 and 7 (E) and GIST 430/654 treated with ML329 (10 μM) at days 7 and 10 (K). Results were analyzed using the Dean/Jett/Fox model and FlowJo 7.0 software. (∗ p < 0.05, ∗∗ p < 0.01; unpaired t test.) Western blots for KIT, MITF, BCL2, and CDK2 are shown after treatment with ML329 in GIST-T1 after 3 days and GIST 430/654 after 10 days (F and L, respectively). β-actin was used as a loading control. Ratios of proteins blotted versus β-actin are shown. All experiments were performed at least three times, and the blots represent the results of several experiments.

Article Snippet: Mouse anti-C-KIT (E1) (clone Ab81), mouse anti-BCL2 (C2), and mouse anti-CDK2 (Clone D-12) were purchased from Santa Cruz Biotechnology, Inc. (Santa Cruz, CA, USA).

Techniques: Expressing, Incubation, Activity Assay, Luciferase, Software, Western Blot, Control