사람 자궁경부암 세포주에서 Cisplatin의 세포독성에 미치는 녹차 폴리페놀의 영향

Abstract

Objective: Green Tea polyphenol(GTP) has been shown to have anti-tumor properties in a wide variety of experimental systems. In this study, we evaluated GTP as a biochemical modulator on the cytotoxic effects of cisplatin in treatment of cancer. Methods: HeLa (Korean Cell Culture Bank) human uterine cervical cancer cell line was used. HeLa Cell was cultured 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 GTP, epigallocatechin gallate(EGCG) and polyphenonⓡ were used for these experiment. Cells were seeded at 1.0×10_(4) cells/well in RPMI1640 media in triplicate wells on a Nunc Labware 96 well flat bottom microculture plate, with or without GTP(50㎍/mL) and at different concentrations of cisplatin (0-100㎍/mL). After incubating the plates at 37℃ in 5% CO2 for 1 days, cell viability was determined using the MTT [3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; thiazolyl blue] 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 HeLa cell lines. Results: The viability of the HeLa cells was decreased to 31% at a 12 ㎍/ml concentration of cisplatin, and to 12 % above 25 ㎍/ml as measured by the MTT assay. However, in the co-treatment with EGCG(50㎍/mL), the cell viability decreased to 47% at 4 ㎍/ml of cisplatin and to 15 % at 6 ㎍/mL of cisplatin in HeLa cells. There are no apparent changes in cisplatin-induced cytotoxicity between EGCG and polyphenonⓡ administration. The levels of p53, caspase-3, Noxa and caspase-8 in HeLa cells were increased by EGCG or polyphenonⓡ treatment. Conclusion: These experiments showed that GTP has a potentiating effect on cisplatin-induced cytotoxicity of HeLa cells. The synergistic activity of GTP might be due to the increased caspase-3 by means of its influence on anti-neoplastic activity of cisplatin.