사람 자궁경부암 세포주에서 Cisplatin의 세포독성에 미치는 녹차풀리페놀의 영향

Abstract

Objective: The aim of this study was to evaluate tea polyphenol(TP) as a biochemical modulator on the cytotoxic effects of cisplatin in uterine cervical cancer cell lines. Methods: HeLa cell and SiHa cells from Korean Cell Culture Bank were cultured in RPMI-1640 medium supplemented with a 10% fetal bovine serum, antibiotics streptomycin and penicillin. Three types of TP-catechin. epigalocatechin galate(EGCG) and polyphenon were used for this study. Cells were seeded at 1×10^(4) cells/well in RPMI1640 media in triplicate wells on a NuncLabware 96 well flat bottom microculture plate, with or without TP(50㎍/㎖) and at different concentrations of cisplatin (0-100㎍/㎖). After incubating the plates at 37℃ in 5% CO_(2) for 1 days, cell viability was determined using the MTT〔3-(4,5- limethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide; thiazolyl blue〕 assay. RT-PCR and western blot method were employed to detect the influence of EGCG and polyphenon on the expressions of p53 gene in HeLa and SiHa cell lines. Results : The viability of HeLa cells was decreased to 31% at 12 ㎍/㎖ concentration of cisplatin, and to 12% above 25㎍/㎖ by the MTT assay. However, in the co-treatment with EGCG(50㎍/㎖), the viability of HeLa cells was decreased to 47% at 4㎍/㎖ of cisplatin and to 15% at 6㎍/㎖ of cisplatin. The viability of SiHa cells was decreased to 81% at 12㎍/㎖ cisplatin, and to 25% above 25㎍/㎖ of cisplatin. However, in EGCG(50㎍/㎖) treated groups, the viability of SiHa cells were decreased to 47% at 6㎍/㎖ of cisplatin and to 15% at 12㎍/㎖ of cisplatin. There is no apparent changes in cisplatin-induced cytotoxicity between EGCG and polyphenon administration. The viability of SiHa cells and HeLa cells were significantly decreased at 6㎍/㎖ of cisplatin when EGCG was administered 2hours before or after cisplatin treatment. But there was no beneficial effect when it was administered with cisplatin simultaneously. After administration of EGCG and polyphenon, p53 protein level was increased in western blot. But p53 expression was unchanged by RT-PCR. Conclusion: This study showed that TP has potentiating effect on cisplatin-induced cytotoxicity ln HeLa and SiHa cell lines especially when it was administered before or after cisplatin treatment. This synergistic activity of TP might be associated with the increased levels of p53 gene.