(-)-Epigallocatechin-3-gallate induces apoptosis in human endometrial adenocarcinoma cells via ROS generation and p38 MAP kinase activation

Murli Manohar, Iram Fatima, Ruchi Saxena, Vishal Chandra, Pushp L. Sankhwar, Anila Dwivedi

Research output: Contribution to journalArticle

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Abstract

(-)-Epigallocatechin-3-gallate (EGCG), the major polyphenol in green tea, has been shown to inhibit carcinogenesis of various tumor types. The aim of this study was to elucidate the antiproliferative potential of EGCG and its mechanism in human endometrial cancer cells (Ishikawa cells) and primary endometrial adenocarcinoma cells. The antiproliferative effect of EGCG was evaluated by cell viability assay. Apoptosis was measured by annexin/propidium iodide staining. Reactive oxygen species (ROS) generation was measured by using 2',7'-dichlorofluorescin diacetate dye. Expression of mitogen-activated protein kinases, proliferation and apoptotic markers were measured by immunoblot analysis. EGCG was found to inhibit proliferation in Ishikawa as well as in primary endometrial adenocarcinoma cells and effectively down-regulated the expression of proliferation markers, i.e., estrogen receptor α, progesterone receptor, proliferating cell nuclear antigen and cyclin D1. EGCG also decreased the activation of ERK and downstream transcription factors fos and jun. EGCG caused apoptotic cell death accompanied by up-regulation of proapoptotic Bax and down-regulation of antiapoptotic protein Bcl2. EGCG induced the cleavage of caspase-3 and poly(ADP-ribose) polymerase, the hallmark of apoptosis. EGCG significantly induced the ROS generation as well as p38 activation in Ishikawa cells, which appeared to be a critical mediator in EGCG-induced apoptosis. The apoptotic effect of EGCG and the p38 activation were blocked by pretreatment of cells with the ROS scavenger N-acetylcysteine. EGCG reduced the glutathione levels, which might be responsible for enhanced ROS generation causing oxidative stress in endometrial cancer cells. Taken together, these results suggest that EGCG inhibits cellular proliferation via inhibiting ERK activation and inducing apoptosis via ROS generation and p38 activation in endometrial carcinoma cells.

Original languageEnglish (US)
Pages (from-to)940-947
Number of pages8
JournalJournal of Nutritional Biochemistry
Volume24
Issue number6
DOIs
StatePublished - Jun 1 2013

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p38 Mitogen-Activated Protein Kinases
Reactive Oxygen Species
Adenocarcinoma
Chemical activation
Apoptosis
Cells
Endometrial Neoplasms
epigallocatechin gallate
Annexins
Oxidative stress
Poly(ADP-ribose) Polymerases
Propidium
Cyclin D1
Proliferating Cell Nuclear Antigen
Acetylcysteine
Polyphenols
Tea
Cell death
Progesterone Receptors
Mitogen-Activated Protein Kinases

All Science Journal Classification (ASJC) codes

  • Endocrinology, Diabetes and Metabolism
  • Biochemistry
  • Molecular Biology
  • Nutrition and Dietetics
  • Clinical Biochemistry

Cite this

(-)-Epigallocatechin-3-gallate induces apoptosis in human endometrial adenocarcinoma cells via ROS generation and p38 MAP kinase activation. / Manohar, Murli; Fatima, Iram; Saxena, Ruchi; Chandra, Vishal; Sankhwar, Pushp L.; Dwivedi, Anila.

In: Journal of Nutritional Biochemistry, Vol. 24, No. 6, 01.06.2013, p. 940-947.

Research output: Contribution to journalArticle

Manohar, Murli ; Fatima, Iram ; Saxena, Ruchi ; Chandra, Vishal ; Sankhwar, Pushp L. ; Dwivedi, Anila. / (-)-Epigallocatechin-3-gallate induces apoptosis in human endometrial adenocarcinoma cells via ROS generation and p38 MAP kinase activation. In: Journal of Nutritional Biochemistry. 2013 ; Vol. 24, No. 6. pp. 940-947.
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