Anti-cancer Potential of ALF, Se NPs, and ALF-Se NPs

Anticancer effect of ALF, Se NPs and ALF-Se NPs was assayed using three human cancer cell lines, including breast cancer cell line (MCF-7), liver cancer cell line (HepG-2) and colon cancer cells (Caco-2). All cell lines were obtained from the American Type Culture Collection (ATCC, USA). MCF7 and HepG2 were maintained in RPMI (Lonza, USA) while Caco2 cell line was cultured in DMEM (Lonza, USA), both media were supplemented with 10% FBS. All cancer cells (5 × 10³ cells/well) were seeded in sterile 96-well plates. After 24 h, serial concentrations of ALF (62.5, 125, 150, 500, 1000 and 2000 ug/ml), Se NPs and ALF-Se NPs (31.25, 62.5, 125, 150, 500 and 1000 ug/ml) as compared to 5-FU (standard reference drug) were incubated with four cancer cell lines for 72 h at 37 °C in 5% CO₂ incubator. MTT method was done as described above. The half maximal inhibitory concentration (IC₅₀) values were calculated using the Graphpad Instat software. Furthermore, cellular morphological changes before and after treatment with ALF, Se NPs and ALF-Se NPs were investigated using phase contrast inverted microscope with a digital camera (Olympus, Japan). Importantly, selectivity index (SI) that defined as the ratio of the IC₅₀ on normal cells versus tumor cells of ALF with or without Se NPs was estimated as IC₅₀-N/IC₅₀^{75 (link)}.