Flow cytometric analysis of the effects of methotrexate and vepesid on the HEp-2 cell cycle

Abstract

Objective: To determine the mechanism of action of Methotrexate and Vepesid on the HEp-2 cells isolated from human laryngeal cancer cells morphologically and flow cytometrically (G1, G2, S and PI). Materials and Methods: The HEp-2 continuous cell line was used. Cultivation of the cells was realized in EMEM medium with 10% fetal bovine serum at an atmosphere of 37şC with 5% $CO_2$. Six different concentrations of Vepesid and Methotrexate were prepared by diluting with deionized water (5 $\mu$g/ml, 50 mg/ml, 500 $\mu$g/ml). The morphological and cell cycle parameters of HEp-2 cells were determined by inverted microscope and flow cytometer respectively. Results: In the morphological examination, Vepesid was found to have a more significant cytopathologic effect on the cells than Methotrexate, whereas in the flow cytometric examination, it was found that whilst Methotrexate stopped the cells at the S and G2 phases, Vepesid did that only at the G1 phase. Conclusion: Both the flow cytometric and cell morphological analysis showed Vepesid to be more effective than Methotrexate on HEp-2 cells. Results of studies conducted show the mechanism of action of these drugs to be dependent on the origin of the cell and on the drug type.Objective: To determine the mechanism of action of Methotrexate and Vepesid on the HEp-2 cells isolated from human laryngeal cancer cells morphologically and flow cytometrically (G1, G2, S and PI). Materials and Methods: The HEp-2 continuous cell line was used. Cultivation of the cells was realized in EMEM medium with 10% fetal bovine serum at an atmosphere of 37şC with 5% $CO_2$. Six different concentrations of Vepesid and Methotrexate were prepared by diluting with deionized water (5 $\mu$g/ml, 50 mg/ml, 500 $\mu$g/ml). The morphological and cell cycle parameters of HEp-2 cells were determined by inverted microscope and flow cytometer respectively. Results: In the morphological examination, Vepesid was found to have a more significant cytopathologic effect on the cells than Methotrexate, whereas in the flow cytometric examination, it was found that whilst Methotrexate stopped the cells at the S and G2 phases, Vepesid did that only at the G1 phase. Conclusion: Both the flow cytometric and cell morphological analysis showed Vepesid to be more effective than Methotrexate on HEp-2 cells. Results of studies conducted show the mechanism of action of these drugs to be dependent on the origin of the cell and on the drug type