Flow Cytometric Analysis of the Effects of Methotrexate and Vepesid on the HEp-2 Cell Cycle
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 37ºC with 5% CO2. Six different concentrations of Vepesid and Methotrexate were prepared by diluting with deionized water (5 µg/ml, 50 mg/ml, 500 µg/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.
Flow Cytometric Analysis of the Effects of Methotrexate and Vepesid on the HEp-2 Cell Cycle
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 37ºC with 5% CO2. Six different concentrations of Vepesid and Methotrexate were prepared by diluting with deionized water (5 µg/ml, 50 mg/ml, 500 µg/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.
