사람 폐암 세포주에서 시스플라틴과 독소루비신의 세포독성에 미치는

Abstract

Objective: Tea polyphenol(TP) has been shown to have anti-tumor properties in a wide variety of experimental systems. In this study, we evaluated TP as a biochemical modulator on the cytotoxic effects of cisplatin or doxorubicin in treatment of lung cancer. Methods: The human lung cancer cells of A549 line was used. A549 Cells(Korean Cell Culture Bank) were grown in RPMI-1640 medium supplemented with 10% (v/v) heat-inactivated fetal bovine serum and antibiotics: 100 units/mL penicillin and 100 ㎍/mL streptomycin. Two types of TP, epigallocatechin galate(EGCG) and polyphenon were used for these experiment. Cells were seeded at 1×104 cells/well in RPMI1640 media in triplicate wells on a Nunc Lab ware 96 well flat bottom microculture plate, with or without TP(50㎍/mL) and at different concentrations of doxorubicin (0-40㎍/mL) or cisplatin(0-100㎍/mL. 1 day after incubation at 37°C in 5% CO2, the cells viability were determined using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide] assay. Western blot method were employed to detect the influence of EGCG and polyphenon on the expressions of p53, capsae 3, caspase 8, and Noxa genes in A549 cell lines. Results: A549 cells viability were decreased to 35% at a 12㎍/mL concentration of cisplatin, and to 14 % above 25 ㎍/mL as measured by the MTT assay. However, in the co-treatment with EGCG(50㎍/mL) or polyphenon(50㎍/mL), the cell viability decreased to 41% at 4㎍/mL of cisplatin and to 16 % at 6㎍/mL of cisplatin in A549 cells. There is no apparent changes in cisplatin-induced cytotoxicity between EGCG and polyphenon administration. A549 cells viability were decreased to 36% at a 6㎍/mL concentration of doxorubicin, but cell viability decreased to 32% at 4㎍/mL of doxorubicin cisplatin and to 16% at 6㎍/mL of cisplatin in EGCG(50㎍/mL) or polyphenon(50㎍/mL) co-treated A549 cells as measured by the MTT assay. There is no apparent changes in doxorubicin-induced cytotoxicity between EGCG and polyphenon administration. The levels of p53, caspase-3, Noxa and caspase-8 in A549 cells were increased by EGCG or polyphenon treatment. Conclusion: These experiments showed that TP has a potentiating effect on doxorubicin- or cisplatin-induced cytotoxicity of A549 cells. The synergistic activity of TP might be due to the induction of apoptosis.