Construction of a cisplatin-induced polyploid giant cancer cell model
OBJECTIVE:To investigate the characteristics and mechanisms of cisplatin-induced polyploid giant cancer cells(PGCCs)in vitro.METHODS:Cultured cells were treated with cisplatin at concentrations of 1.5,3,6,12,and 24 μg/mL for 3,4,and 5 days.DNA content was analyzed by flow cytometry to determine the optimal combination of concentration and duration for PGCC induction in A549,HepG2,and SK-OV-3 cells.Morphological changes and DNA nuclear area were examined through morphological observation and Giemsa staining.Expression of stemness genes(Nanog,Sox-2,OCT-4,c-Myc)was evaluated by RT-qPCR,while stemness surface markers(CD44,CD 133)were analyzed by flow cytometry.Cell senescence was assessed by β-galactosidase staining.After 72-hour treatment with cisplatin at concentrations ranging from 0.75 to 96 μg/mL,cell viability was measured by the MTT assay.RESULTS:Treatment with 3 μg/mL cisplatin for 3 d effectively induced polyploidy(＞4N)in all three cell lines.The induced cells exhibited significantly increased cell and nuclear areas;varying degrees of upregulation in stemness markers CD44 and CD 133,along with partial increase in stemness gene expression;partial senescence phenotype in A549 cells;and enhanced cisplatin tolerance in A549 and SK-OV-3 PGCCs compared to controls.CONCLUSION:The cisplatin-induced PGCC model in A549 cells demonstrated enlarged cell and nuclear areas,expressed stemness genes(Nanog,Sox-2,OCT-4),and showed increased cisplatin tolerance.The model may be useful for investigating,drug resistance mechanisms.
cisplatinpolyploid giant cancer cellA549 cellsHepG2 cellsSK-OV-3 cells
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