TY  - EJOU
AU  - Papakonstantinou, Dimitrios 
AU  - Vardas, Vasileios 
AU  - Varouhaki, Despoina M. 
AU  - Kotzamouratoglou, Aikaterini 
AU  - Mangani, Karolina 
AU  - Ju, lia A. 
AU  - Alix-Panabières, Catherine 
AU  - Martin, Stuart S. 
AU  - Athanassopoulos, Constantinos M. 
AU  - Kallergi, Galatea 

TI  - The Impact of Combretastatin A-4 on Cancer Cells and Circulating Tumor Cells (CTCs): A Multi-Assay Approach
T2  - Oncology Research

PY  - 
VL  - 
IS  - 
SN  - 1555-3906

AB  - <b>Objectives:</b> Combretastatin A-4 (CA-4) is a microtubule-disrupting agent with established anti-tumor properties. This study aimed to evaluate the effects of CA-4 on key metastatic traits of cancer cells, including migration, clonogenic potential, cytoskeletal protein expression, and microtentacle (McTN) formation, using multiple cancer cell models, including the colon patient-derived circulating tumor cell line CTC-MCC-41. <b>Methods:</b> H1299 (non-small cell lung cancer), MDA-MB-231 (triple-negative breast cancer), HT-29 (colorectal cancer), and CTC-MCC-41 (derived from the blood of a colon cancer patient) cells were treated with CA-4 (10 μM) for 24 and 48 h. Colony formation was assessed with a clonogenic assay. Migration assays (Boyden chamber) evaluated cell motility, while cell viability was determined via MTT assay. Western blot analysis examined vimentin and α/β-tubulin expression. TetherChip assay analyzed McTN formation using wheat germ agglutinin (WGA) staining. <b>Results:</b> CA-4 treatment significantly reduced cell viability, colony formation, and migration across all tested cancer cell lines. Western blot analysis revealed a marked reduction in vimentin and α/β-tubulin expression after 48 h of treatment, indicating disruption of cytoskeletal integrity. In addition, TetherChip analysis demonstrated a pronounced decrease in McTN formation following CA-4 exposure, suggesting inhibition of cytoskeletal protrusions associated with metastatic dissemination. <b>Conclusions:</b> CA-4 effectively impairs multiple cancer cell functions related to metastatic progression, including proliferation, migration, cytoskeletal organization, and McTN formation. These findings highlight the potential of CA-4 as a microtubule-targeting agent with anti-metastatic activity, particularly in circulating tumor cells.
KW  - Combretastatin A-4; circulating tumor cells; microtubules; microtentacles; metastasis

DO  - 10.32604/or.2026.085665