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  • Time-dependent intracellular trafficking of FITC-conjugated epigallocatechin-3-O-gallate in L-929 cells.

Time-dependent intracellular trafficking of FITC-conjugated epigallocatechin-3-O-gallate in L-929 cells.

Bioorganic & medicinal chemistry (2008-10-28)
Dong-Wook Han, Kazuaki Matsumura, Bongju Kim, Suong-Hyu Hyon
ABSTRACT

Many in vitro studies about green tea polyphenol, (-)-epigallocatechin-3-O-gallate (EGCG) focused on its pro-apoptotic and anti-proliferative effects on various types of cancer cells, while less attention has been paid to its incorporation into the cytoplasm and nuclear translocation. This study concentrated on the time-dependent intracellular trafficking of EGCG in L-929 cells. EGCG was conjugated with fluorescein-4-isothiocyanate (FITC) via the 3''-OH or 5''-OH group, as confirmed by NMR analysis, and then treated to either suspended or cultured cells. Confocal microscopic observations revealed that FITC-EGCG was clearly seen onto the membrane of suspended cells as well as into the cytoplasm and nucleus within 1h. As an increase in treatment time, it concentrated on the nucleus and then was located at any places of the cells. The cellular uptake of FITC-EGCG in cultured cells was not observed until 1h of culture, but started to be observed after at least 2h. These results imply that although the cellular sensitivity and response to EGCG would be different from those of FITC-EGCG, it would be incorporated into the cytoplasm of cells and further be translocated into the nucleus in a time-dependent manner.

MATERIALS
Product Number
Brand
Product Description

Sigma-Aldrich
(−)-Epigallocatechin gallate, ≥80% (HPLC), from green tea
Sigma-Aldrich
(−)-Epigallocatechin gallate, ≥95%
Sigma-Aldrich
Fluorescein 5(6)-isothiocyanate, ≥90% (HPLC)
Sigma-Aldrich
Fluorescein isothiocyanate isomer I–Celite®, suitable for fluorescent labeling techniques
Sigma-Aldrich
Fluorescein isothiocyanate isomer I, ≥97.5% (HPLC)
Sigma-Aldrich
Fluorescein isothiocyanate isomer I, suitable for protein labeling, ≥90% (HPLC), powder