In vitro effects and mathematical modelling of CTCE‐9908 (a chemokine receptor 4 antagonist) on melanoma cell survival

Abstract

AbstractCTCE‐9908, a CXC chemokine receptor 4 (CXCR4) antagonist, prevents CXCR4 phosphorylation and inhibits the interaction with chemokine ligand 12 (CXCL12) and downstream signalling pathways associated with metastasis. This study evaluated the in vitro effects of CTCE‐9908 on B16 F10 melanoma cells with the use of mathematical modelling. Crystal violet staining was used to construct a mathematical model of CTCE‐9908 B16 F10 (melanoma) and RAW 264.7 (non‐cancerous macrophage) cell lines on cell viability to predict the half‐maximal inhibitory concentration (IC50). Morphological changes were assessed using transmission electron microscopy. Flow cytometry was used to assess changes in cell cycle distribution, apoptosis via caspase‐3, cell survival via extracellular signal‐regulated kinase1/2 activation, CXCR4 activation and CXCL12 expression. Mathematical modelling predicted IC50 values from 0 to 100 h. At IC50, similar cytotoxicity between the two cell lines and ultrastructural morphological changes indicative of cell death were observed. At a concentration 10 times lower than IC50, CTCE‐9908 induced inhibition of cell survival (p = 0.0133) in B16 F10 cells but did not affect caspase‐3 or cell cycle distribution in either cell line. This study predicts CTCE‐9908 IC50 values at various time points using mathematical modelling, revealing cytotoxicity in melanoma and non‐cancerous cells. CTCE‐9908 significantly inhibited melanoma cell survival at a concentration 10 times lower than the IC50 in B16 F10 cells but not RAW 264.7 cells. However, CTCE‐9908 did not affect CXCR4 phosphorylation, apoptosis,\ or cell cycle distribution in either cell line.