Journal: Neoplasia (New York, N.Y.)
Article Title: A MYC family switch: L-MYC drives and maintains neuroendocrine lineage programs in prostate cancer
doi: 10.1016/j.neo.2026.101307
Figure Lengend Snippet: MYCL reduces proliferation while promoting migration and cytoskeletal remodeling and decreasing cell adhesion in prostate cancer cells (A) Proliferation assay. Real-time Incucyte analysis showing cell proliferation of C4-2B control (Ctrl) and C4-2B-MYCL cells measured as confluence (%) over time. Quantification using area under the curve (AUC) demonstrates reduced proliferative capacity following MYCL overexpression. (B) Proliferation assay. Real-time Incucyte analysis showing cell proliferation of PC3 control (Ctrl) and PC3-MYCL cells measured as confluence (%) over time. Quantification using area under the curve (AUC) demonstrates reduced proliferative capacity following MYCL overexpression. (C) Left: Cell cycle analysis of C4-2B and PC3 cells following MYCL overexpression. In PC3 cells, MYCL reduces the G1 population and increases S-phase, indicating altered cell cycle progression, whereas changes in C4-2B cells are minimal. Right: Apoptosis analysis by Annexin V/7-AAD staining. MYCL overexpression in PC3 cells increases the early apoptotic population, with little or no significant change in C4-2B cells. (D) Cell adhesion assay. Cell adhesion was assessed by crystal violet staining and quantified by measuring absorbance at 595 nm at 18, 24, 48, and 72 h following seeding. MYCL-overexpressing C4-2B cells exhibited significantly decreased adhesion compared with control cells. Representative phase-contrast images acquired 48 h after seeding show reduced cell attachment and increased cell clustering in MYCL-expressing cells. (E) Molecular validation. RT-qPCR analysis of adhesion-related genes (ITGB1, ITGAV) in control (Ctrl) and MYCL-overexpressing C4-2B, LNCaP and PC3 cells. (F) Migration assay. Wound-healing analysis measuring relative wound density (%) over time demonstrates moderately enhanced migratory capacity in PC3-MYCL cells. AUC quantification confirms increased migration upon MYCL expression. (G) Cytoskeletal transcriptional programs. Heatmap showing differential expression of genes meeting thresholds of |log₂FC| ≥ 0.5 and adjusted p-value (FDR) < 0.05. Differentially regulated genes are associated with Ephrin-EPH signaling, Rho-Rac signaling, cytoskeletal organization, cell-cell junctions, extracellular matrix (ECM) interactions, and epithelial-mesenchymal transition (EMT) regulators, indicating MYCL-driven cytoskeletal remodeling signatures.
Article Snippet: For live-cell proliferation analysis, cells were seeded in 96-well plates and monitored using the IncuCyte® Live-Cell Analysis System (Sartorius).
Techniques: Migration, Proliferation Assay, Control, Over Expression, Cell Cycle Assay, Staining, Cell Adhesion Assay, Cell Attachment Assay, Expressing, Biomarker Discovery, Quantitative RT-PCR, Quantitative Proteomics