Cytotoxic effects of synthetic alkyl-phospholipid (Erufosine) in combination with chemotherapy (5-FU) against colorectal cancer cells

Abstract

Background: Colorectal cancer (CRC) is one of the most commonly diagnosed malignancy. An approach used in treatment of this cancer is combinational treatment where anticancer compounds are combined for cure. Exploring the novel anticancer compounds for potential usage as combinational approach is burning domain. In this regard, alkyl-phospholipids are novel agents and erufosine is a 3^rd generation alkyl-phospholipid compound that interact with surface membrane of cell and induces antineoplastic effects in malignant cells.

Material and Methods: Toxic effects of erufosine and 5-FU (5-FU) were identified against the CRC cell lines. For this purpose, the cells were exposed to various concentrations of the test compounds for 24-72 h and cell viability was assessed by MTT dye reduction assay. Afterwards, the cells were exposed to three distinct concentrations of the test compounds (IC₂₅, IC₅₀, IC₇₅) for 48 h and expressional modulations in cell cycle regulator (P21) and stress marker (GADD45A) were identified by qRT-PCR methodology. For both MTT assay and qRT-PCR analysis, the cells were exposed to selected compounds as single agent or combination in parallel to avoid any time-lapse related differences between the experimental findings.

Results: Erufosine and 5-FU induced substantial anti-proliferative effects in SW480, SW620, HCT116 CRC cells. The inhibitory effects of erufosine were more pronounced in comparison to 5-FU. Furthermore, synergistic anti-proliferative effects were observed when erufosine was combined with 5-FU. In addition, qRT-PCR data showed noteworthy potential of the test compounds to induce expression of P21 and GADD45A genes at mRNA levels in the cells. Precisely, combination of erufosine and 5-FU induced P21 gene more prominently in the cells, especially in metastatic CRC cells (SW620).    

Conclusion: Erufosine and 5-FU bear substantial cytotoxic potential against the CRC cells. Combination of erufosine and 5-FU showed synergistic anti-proliferative effects in the cells. The compounds up-regulate expression of a cell cycle inhibitor and cell stress marker (P21 and GADD45A gene) in CRC cells.