Inhibition of TPA-induced protein kinase C and transcription activator protein-1 binding activities by theaflavin-3,3'-digallate from black tea in NIH3T3 cells

Yen Chou Chen, Yu Chih Liang, Shoei Yn Lin-Shiau, Chi Tang Ho, Jen Kun Lin

Research output: Contribution to journalArticle

49 Citations (Scopus)

Abstract

Tea is one of the most popular beverages in the world. Several reports have shown that both green tea and black tea were able to inhibit tumor cell proliferation in animal models. In this study, we investigated the inhibitory effects of black tea polyphenols including theaflavin (TF-1), the mixture (TF-2) of theaflavin-3-gallate (TF-2a), and theaflavin-3'-gallate (TF-2b), theaflavin-3,3'-digallate (TF-3), thearubigin (TR), and a major green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) on 12-O- tetradecanoylphorbol 13-acetate (TPA)-induced protein kinase C (PKC) and transcription activator protein-1 (AP-1) binding activities in NIH3T3 cells. On analysis of PKC activity with partial purified preparation, TPA (100 ng/mL) treatment was able to elevate membrane-associated PKC activity ~3- fold, and treatment with TF-3 (20 μM) and EGCG (20 μM) showed 94.5% and 9.4% suppression on TPA-induced PKC activity, respectively. Translocation of PKCα protein from cytosol to membrane was detected in TPA-treated NIH3T3 cells, and TF-3 was able to block its translocation. By in vitro kinase assay using myelin basic protein (MBP) as a PKC-specific substrate, we found that TPA treatment was able to increase PKC kinase activity by detection of phosphorylated MBP protein and TF-3 showed strongest inhibitory effect on its phosphorylation while EGCG was shown to be less effective. We also analyzed the AP-1 binding activity by electrophoretic mobility shift assay and c-Jun gene expression by northern blot and western blot, the results showed that TF-3 is the most potent inhibitor on TPA-induced AP-1 binding activity and c- Jun gene expression among these five tea polyphenols. Our results might provide new molecular basis for understanding the inhibitory effects of tea polyphenols on TPA-mediated tumor promotion.

Original languageEnglish
Pages (from-to)1416-1421
Number of pages6
JournalJournal of Agricultural and Food Chemistry
Volume47
Issue number4
DOIs
Publication statusPublished - Apr 1999
Externally publishedYes

Fingerprint

theaflavins
black tea
Transcription Factor AP-1
protein kinase C
Tetradecanoylphorbol Acetate
Tea
Transcription
Protein Binding
Protein Kinase C
Acetates
acetates
Polyphenols
epigallocatechin
polyphenols
jun Genes
proteins
cells
Myelin Basic Protein
Gene expression
green tea

Keywords

  • AP-1 binding activity
  • Black tea
  • PKC
  • Theaflavin-3,3'-digallate
  • TPA
  • Tumor promotion

ASJC Scopus subject areas

  • Agricultural and Biological Sciences (miscellaneous)
  • Food Science
  • Chemistry (miscellaneous)

Cite this

@article{0f90aa567fc2486a8e39e925f4e36503,
title = "Inhibition of TPA-induced protein kinase C and transcription activator protein-1 binding activities by theaflavin-3,3'-digallate from black tea in NIH3T3 cells",
abstract = "Tea is one of the most popular beverages in the world. Several reports have shown that both green tea and black tea were able to inhibit tumor cell proliferation in animal models. In this study, we investigated the inhibitory effects of black tea polyphenols including theaflavin (TF-1), the mixture (TF-2) of theaflavin-3-gallate (TF-2a), and theaflavin-3'-gallate (TF-2b), theaflavin-3,3'-digallate (TF-3), thearubigin (TR), and a major green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) on 12-O- tetradecanoylphorbol 13-acetate (TPA)-induced protein kinase C (PKC) and transcription activator protein-1 (AP-1) binding activities in NIH3T3 cells. On analysis of PKC activity with partial purified preparation, TPA (100 ng/mL) treatment was able to elevate membrane-associated PKC activity ~3- fold, and treatment with TF-3 (20 μM) and EGCG (20 μM) showed 94.5{\%} and 9.4{\%} suppression on TPA-induced PKC activity, respectively. Translocation of PKCα protein from cytosol to membrane was detected in TPA-treated NIH3T3 cells, and TF-3 was able to block its translocation. By in vitro kinase assay using myelin basic protein (MBP) as a PKC-specific substrate, we found that TPA treatment was able to increase PKC kinase activity by detection of phosphorylated MBP protein and TF-3 showed strongest inhibitory effect on its phosphorylation while EGCG was shown to be less effective. We also analyzed the AP-1 binding activity by electrophoretic mobility shift assay and c-Jun gene expression by northern blot and western blot, the results showed that TF-3 is the most potent inhibitor on TPA-induced AP-1 binding activity and c- Jun gene expression among these five tea polyphenols. Our results might provide new molecular basis for understanding the inhibitory effects of tea polyphenols on TPA-mediated tumor promotion.",
keywords = "AP-1 binding activity, Black tea, PKC, Theaflavin-3,3'-digallate, TPA, Tumor promotion",
author = "Chen, {Yen Chou} and Liang, {Yu Chih} and Lin-Shiau, {Shoei Yn} and Ho, {Chi Tang} and Lin, {Jen Kun}",
year = "1999",
month = "4",
doi = "10.1021/jf981099k",
language = "English",
volume = "47",
pages = "1416--1421",
journal = "Journal of Agricultural and Food Chemistry",
issn = "0021-8561",
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TY - JOUR

T1 - Inhibition of TPA-induced protein kinase C and transcription activator protein-1 binding activities by theaflavin-3,3'-digallate from black tea in NIH3T3 cells

AU - Chen, Yen Chou

AU - Liang, Yu Chih

AU - Lin-Shiau, Shoei Yn

AU - Ho, Chi Tang

AU - Lin, Jen Kun

PY - 1999/4

Y1 - 1999/4

N2 - Tea is one of the most popular beverages in the world. Several reports have shown that both green tea and black tea were able to inhibit tumor cell proliferation in animal models. In this study, we investigated the inhibitory effects of black tea polyphenols including theaflavin (TF-1), the mixture (TF-2) of theaflavin-3-gallate (TF-2a), and theaflavin-3'-gallate (TF-2b), theaflavin-3,3'-digallate (TF-3), thearubigin (TR), and a major green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) on 12-O- tetradecanoylphorbol 13-acetate (TPA)-induced protein kinase C (PKC) and transcription activator protein-1 (AP-1) binding activities in NIH3T3 cells. On analysis of PKC activity with partial purified preparation, TPA (100 ng/mL) treatment was able to elevate membrane-associated PKC activity ~3- fold, and treatment with TF-3 (20 μM) and EGCG (20 μM) showed 94.5% and 9.4% suppression on TPA-induced PKC activity, respectively. Translocation of PKCα protein from cytosol to membrane was detected in TPA-treated NIH3T3 cells, and TF-3 was able to block its translocation. By in vitro kinase assay using myelin basic protein (MBP) as a PKC-specific substrate, we found that TPA treatment was able to increase PKC kinase activity by detection of phosphorylated MBP protein and TF-3 showed strongest inhibitory effect on its phosphorylation while EGCG was shown to be less effective. We also analyzed the AP-1 binding activity by electrophoretic mobility shift assay and c-Jun gene expression by northern blot and western blot, the results showed that TF-3 is the most potent inhibitor on TPA-induced AP-1 binding activity and c- Jun gene expression among these five tea polyphenols. Our results might provide new molecular basis for understanding the inhibitory effects of tea polyphenols on TPA-mediated tumor promotion.

AB - Tea is one of the most popular beverages in the world. Several reports have shown that both green tea and black tea were able to inhibit tumor cell proliferation in animal models. In this study, we investigated the inhibitory effects of black tea polyphenols including theaflavin (TF-1), the mixture (TF-2) of theaflavin-3-gallate (TF-2a), and theaflavin-3'-gallate (TF-2b), theaflavin-3,3'-digallate (TF-3), thearubigin (TR), and a major green tea polyphenol (-)-epigallocatechin-3-gallate (EGCG) on 12-O- tetradecanoylphorbol 13-acetate (TPA)-induced protein kinase C (PKC) and transcription activator protein-1 (AP-1) binding activities in NIH3T3 cells. On analysis of PKC activity with partial purified preparation, TPA (100 ng/mL) treatment was able to elevate membrane-associated PKC activity ~3- fold, and treatment with TF-3 (20 μM) and EGCG (20 μM) showed 94.5% and 9.4% suppression on TPA-induced PKC activity, respectively. Translocation of PKCα protein from cytosol to membrane was detected in TPA-treated NIH3T3 cells, and TF-3 was able to block its translocation. By in vitro kinase assay using myelin basic protein (MBP) as a PKC-specific substrate, we found that TPA treatment was able to increase PKC kinase activity by detection of phosphorylated MBP protein and TF-3 showed strongest inhibitory effect on its phosphorylation while EGCG was shown to be less effective. We also analyzed the AP-1 binding activity by electrophoretic mobility shift assay and c-Jun gene expression by northern blot and western blot, the results showed that TF-3 is the most potent inhibitor on TPA-induced AP-1 binding activity and c- Jun gene expression among these five tea polyphenols. Our results might provide new molecular basis for understanding the inhibitory effects of tea polyphenols on TPA-mediated tumor promotion.

KW - AP-1 binding activity

KW - Black tea

KW - PKC

KW - Theaflavin-3,3'-digallate

KW - TPA

KW - Tumor promotion

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U2 - 10.1021/jf981099k

DO - 10.1021/jf981099k

M3 - Article

C2 - 10563991

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VL - 47

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JO - Journal of Agricultural and Food Chemistry

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SN - 0021-8561

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