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Apoptosis

, Volume 16, Issue 1, pp 75–85 | Cite as

Apoptosis of human fibrosarcoma HT-1080 cells by epigallocatechin-3-O-gallate via induction of p53 and caspases as well as suppression of Bcl-2 and phosphorylated nuclear factor-κB

  • Mi Hee Lee
  • Dong-Wook Han
  • Suong-Hyu Hyon
  • Jong-Chul ParkEmail author
Original Paper

Abstract

Animal tumor bioassays and in vitro cell culture systems have demonstrated that epigallocatechin-3-O-gallate (EGCG), the predominant catechin in green tea, possesses anti-proliferative and pro-apoptotic effects on various cancer cells and tumors. In this study, we investigated the effects of EGCG on cell growth, cell cycle progression, and apoptosis in human fibrosarcoma HT-1080 cells. The involvement of p53, Bcl-2, Bax, caspases, and nuclear factor-κB (NF-κB) was examined as a mechanism for the anti-cancer activity of EGCG. Time-dependent intracellular trafficking of EGCG was also determined using fluorescein isothiocyanate (FITC)-conjugated EGCG (FITC-EGCG). Our data show that EGCG treatment caused dose-dependent cell growth inhibition, cell cycle arrest at the G0/G1 phase, and DNA fragmentation suggesting the induction of apoptosis in HT-1080 cells. Immunoblot analysis revealed that the expression of p53, caspase-7 and -9 as well as the ratio of Bax/Bcl-2 protein increased significantly with higher EGCG concentrations and longer incubation times. Moreover, expression of phosphorylated NF-κB/p65 in HT-1080 cells was inhibited by EGCG treatment in a dose-dependent manner, while that of unphosphorylated NF-κB/p65 remained unaffected. Here we also reveal time-dependent internalization of FITC-EGCG into the cytosol of HT-1080 cells and its subsequent nuclear translocation. These results suggest that EGCG may interrupt exogenous signals directed towards genes involved in proliferation and cell cycle progression. Taken together, our data indicate that HT-1080 apoptosis may be mediated through the induction of p53 and caspases by the pro-oxidant activity of internalized EGCG, as well as suppression of Bcl-2 and phosphorylated NF-κB by the antioxidant activity of EGCG.

Keywords

Epigallocatechin-3-O-gallate Human fibrosarcoma cells Apoptosis p53 Caspases Nuclear factor-κB 

Notes

Acknowledgments

This research was partly supported by the Korea Healthcare Technology R&D Project, Ministry for Health, Welfare & Family Affairs, Republic of Korea (A085136) and by a grant from the Fundamental R&D Program for Core Technology of Materials funded by the Ministry of Knowledge Economy, Republic of Korea (K0006028).

Supplementary material

10495_2010_548_MOESM1_ESM.doc (66 kb)
Supplementary material 1 (DOC 66 kb)

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Copyright information

© Springer Science+Business Media, LLC 2010

Authors and Affiliations

  • Mi Hee Lee
    • 1
    • 2
  • Dong-Wook Han
    • 3
  • Suong-Hyu Hyon
    • 4
  • Jong-Chul Park
    • 1
    • 2
    Email author
  1. 1.Cellbiocontrol Laboratory, Department of Medical Engineering Yonsei University College of MedicineSeoulKorea
  2. 2.Brain Korea 21 Project for Medical ScienceYonsei University College of MedicineSeoulKorea
  3. 3.Department of Nanomedical EngineeringCollege of Nanoscience & Nanotechnology, Pusan National UniversityBusanKorea
  4. 4.Department of Medical Simulation EngineeringResearch Center for Nano Medical Engineering, Institute for Frontier Medical Sciences, Kyoto UniversityKyotoJapan

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