(-)-Epigallocatechin-3-gallate induced apoptosis - dissociation of c-FLIP/Ku70 complex - in gastric cancer cells

Sep 24, 2026

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Anti-cancer activity of (-)-epigallocatechin-3-gallate (EGCG) involves induction of apoptosis and suppression of proliferation, including inhibition of histone deacetylase. The cytoplasmic accumulation of stabilized c-FLIP/Ku70 complexes can block apoptosis by disrupting the extrinsic apoptosis pathway. This study assesses whether EGCG promotes apoptosis in gastric cancer cells by dissociating the c-FLIP/Ku70 complex using EGCG or MG149 treatments and multiple apoptosis and protein analyses.

LanguageEnglish

CategoryHealthcare

FormatPDF

Pages11

Reading Time28 min

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Abstract

Introduction

Methods and assessments (flow cytometry, RT-PCR, western blot, immunofluorescence, fractionation, co-immunoprecipitation)

Results (P53/P21 up-regulation, c-Myc/Cyclin D1 down-regulation, cell cycle arrest, c-FLIP/Ku70 dissociation and Ku70 translocation)

Conclusion (role of EGCG as an alternative to conventional HDAC inhibitors and therapeutic combinations)

DOI: 10. 1111/jcmm.17873 OR I G I NAL ART I C L E (-)-Epigallocatechin-3-gallate induced apoptosis by dissociation of c-FLIP/Ku70 complex in gastric cancer cells Mahtab Shahriari Felordi1 | Mehdi Alikhani1 | Zahra Farzaneh2 | Mahmoud Alipour Choshali2 | Marzieh Ebrahimi1,2 | Hamidreza Aboulkheyr Es3 | Abbas Piryaei4,5 | Mustapha Najimi6 | Massoud Vosough1,7 1Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran 2Department of Stem Cells and Developmental Biology, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran, Iran 3School of Biomedical Engineering, University of Technology Sydney, Sydney, New South Wales, Australia 4Department of Biology and Anatomical Sciences, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran 5Department of Tissue Engineering and Applied Cell Sciences, School of Advanced Technologies in Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran 6Laboratory of Pediatric Hepatology and Cell Therapy, Institute of Experimental and Clinical Research (IREC), Brussels, Belgium 7Experimental Cancer Medicine, Department of Laboratory Medicine, Karolinska Institute, Stockholm, Sweden Correspondence Massoud Vosough, Department of Regenerative Medicine, Cell Science Research Center, Royan Institute for Stem Cell Biology and Technology, ACECR, Tehran 16635148, Iran. [Email:](Email: masvos@royaninstitute.org)[ masvos@royaninstitute.org](Email: masvos@royaninstitute.org) Mustapha Najimi, Laboratory of Pediatric Hepatology and Cell Therapy, Institute of Experimental and Clinical Research (IREC), UCLouvain, Brussels, Belgium. [Email:](Email: mustapha. najimi@uclouvain.be)[ mustapha. najimi@uclouvain.be](Email: mustapha. najimi@uclouvain.be) Funding information Bahar Tashkhis Teb Co, Grant/Award Number: R.98005; Royan Institute, Grant/ Award Number: R1398-0335 Abstract Anti-cancer properties of (-)-epigallocatechin-3-gallate (EGCG) are mediated via apoptosis induction, as well as inhibition of cell proliferation and histone deacetylase. Accumulation of stabilized cellular FLICE-inhibitory protein (c-FLIP)/Ku70 complex in the cytoplasm inhibits apoptosis through interruption of extrinsic apoptosis pathway. In this study, we evaluated the anti-cancer role of EGCG in gastric cancer (GC) cells through dissociation of c-FLIP/Ku70 complex. MKN-45 cells were treated with EGCGor its antagonist MG149 for 24 h. Apoptosis was evaluated by flow cytometry and quantitative RT-PCR. Protein expression ofc-FLIP and Ku70 was analysed using western blot and immunofluorescence. Dissociation of c-FLIP/Ku70 complex as well as Ku70 translocation were studied by sub-cellular fractionation and co-immunoprecipitation. EGCG induced apoptosis in MKN-45 cells with substantial up-regulation of P53 and P21, down-regulation of c-Myc and Cyclin D1 as well as cell cycle arrest in S and G2/M check points. Moreover, EGCG treatment suppressed the expression of c-FLIP and Ku70, decreased their interaction while increasing the Ku70 nuclear content. By dissociating the c-FLIP/Ku70 complex, EGCG could be an alternative component to the conventional HDAC inhibitors in order to induce apoptosis in GC cells. Thus, its combination with other cancer therapy protocols could result in a better therapeutic outcome. This is an open access article under the terms of the Creative Commons Attribution License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited. © 2023 The Authors. Journal of Cellular and Molecular Medicine published by Foundation for Cellular and Molecular Medicine and John Wiley & Sons Ltd. K E Y WO R D S apoptosis induction, c-FLIP/Ku70 complex, EGCG treatment, epigenetic modification, gastric cancer, histone deacetylation 1 | INTRODUCTION Gastric cancer (GC) is the fifth most

FAQs

How does EGCG promote apoptosis in gastric cancer cells in this study?

EGCG induces apoptosis in MKN-45 cells by dissociating the c-FLIP/Ku70 complex, which disrupts the inhibition of apoptosis mediated by that complex.

What experimental approaches were used to evaluate apoptosis and gene expression?

Apoptosis was assessed by flow cytometry, and quantitative RT-PCR was used to evaluate apoptosis-related molecular changes. Protein levels were analyzed by western blot and immunofluorescence.

What cell-cycle effects were reported after EGCG treatment?

EGCG treatment caused cell cycle arrest at S and G2/M checkpoints, alongside molecular changes including increased P53 and P21 and decreased c-Myc and Cyclin D1.

Sep 24, 2026

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