Mohammad Panji1, Vahideh Behmard2, Zahra Zare3, Monireh Malekpour4, Hasan Nejadbiglari5, Saeede Yavari6, Tina Nayerpour Dizaj7, Azadeh Safaeian8, Narges Maleki9, Mojtaba Abbasi10, Omid Abazari11, Maryam Shabanzadeh12, Parisa Khanicheragh13. 1. Department of Molecular Medicine, School of Advanced Technologies in Medicine, Tehran University of Medical Sciences, Tehran, Iran. 2. Student Research Committee, Department of Midwifery, School of medical, Gonabad University of Medical Sciences, Gonabad, Iran. 3. Department of Biology, Farhangian University, Tehran, Iran. 4. Department of Immunology, School of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran. 5. Department of Nursing, Sirjan Branch, Islamic Azad University, Sirjan, Iran. 6. Department of Biology, Science and Research Branch, Islamic Azad University, Tehran, Iran. 7. Department of Medical Biotechnology, Faculty of Modern Sciences, Tabriz University of Medical Sciences, Tabriz, Iran. 8. Department of Physiology, Faculty of Medicine, Shahid Sadoughy University of Medical Sciences, Yazd, Iran. 9. Department of Cellular and Molecular Biology, Faculty of Biological Sciences, Islamic Azad University-Tehran North Branch, Tehran, Iran. 10. Veterinary Medicine, Faculty of Veterinary Medicine, Shahrekord Branch, Islamic Azad University, Shahrekord, Iran; Reproductive Biotechnology Research Center, Avicenna Research Institute, ACECR, Tehran, Iran. 11. Department of Clinical Biochemistry, School of Medicine, Shahid Sadoughi University of Medical Sciences and Health Services, Yazd, Iran. 12. Department of Medical Radiation, Faculty of Engineering, Islamic Azad University, Science and Research Branch, Tehran, Iran. Electronic address: taraneh400@gmail.com. 13. Department of Clinical Biochemistry, Lorestan University of Medical Sciences, Khorramabad, Iran. Electronic address: khany.parisa@gmail.com.
Abstract
BACKGROUND: Transforming growth factor-β (TGF-β)-induced Epithelial-to-mesenchymal transition (EMT) process is a fundamental target for preventing cervical cancer cells' progression and invasion. Green tea and its principal active substance, Epigallocatechin-3-gallate (EGCG), demonstrate anti-tumor activities in various tumor cells. METHODS: The cell viability of two cervical cancer cell lines, Hela and SiHa, in the experimental groups was examined employing the MTT method, and ROS generation was probed applying 2',7'-dichlorofluorescein diacetate-based assay. The Smad signaling and EMT process was evaluated utilizing western blot analysis and quantitative real-time polymerase chain reaction (qRT-PCR). Chromatin immunoprecipitation (ChIP) and Smad binding element (SBE)-luciferase assays were employed to measure Smad-DNA interaction and Smad transcriptional activity, respectively. RESULTS: EGCG (0-100 μmol/L) and green tea extract (0-250 μg/ml) suppressed the viability of cancer cells in a dose-dependent manner (p<0.01). Our conclusions affirmed that pre-incubation with green tea extract (80 μg/ml) and EGCG (60 μmol/L) significantly reversed the impacts of TGF-β in Hela and SiHa cells by decreasing Vimentin, ZEB, Slug, Snail, and Twist and increasing E-cadherin expression. The molecular mechanism of green tea extract and EGCG for TGF-β-induced EMT inhibition interfered with ROS generation and Smad signaling. Green tea extract and EGCG could significantly decrease ROS levels, the phosphorylation of Smad2/3, the translocation, DNA binding, and activity of Smads in cervical cancer cell lines treated with TGF-β1 (p<0.01). CONCLUSION: EGCG and green tea extract suppressed TGF-β-induced EMT in Hela and SiHa cells, and the underlying molecular mechanism may be related to the ROS generation and Smad signaling pathway.
BACKGROUND: Transforming growth factor-β (TGF-β)-induced Epithelial-to-mesenchymal transition (EMT) process is a fundamental target for preventing cervical cancer cells' progression and invasion. Green tea and its principal active substance, Epigallocatechin-3-gallate (EGCG), demonstrate anti-tumor activities in various tumor cells. METHODS: The cell viability of two cervical cancer cell lines, Hela and SiHa, in the experimental groups was examined employing the MTT method, and ROS generation was probed applying 2',7'-dichlorofluorescein diacetate-based assay. The Smad signaling and EMT process was evaluated utilizing western blot analysis and quantitative real-time polymerase chain reaction (qRT-PCR). Chromatin immunoprecipitation (ChIP) and Smad binding element (SBE)-luciferase assays were employed to measure Smad-DNA interaction and Smad transcriptional activity, respectively. RESULTS:EGCG (0-100 μmol/L) and green tea extract (0-250 μg/ml) suppressed the viability of cancer cells in a dose-dependent manner (p<0.01). Our conclusions affirmed that pre-incubation with green tea extract (80 μg/ml) and EGCG (60 μmol/L) significantly reversed the impacts of TGF-β in Hela and SiHa cells by decreasing Vimentin, ZEB, Slug, Snail, and Twist and increasing E-cadherin expression. The molecular mechanism of green tea extract and EGCG for TGF-β-induced EMT inhibition interfered with ROS generation and Smad signaling. Green tea extract and EGCG could significantly decrease ROS levels, the phosphorylation of Smad2/3, the translocation, DNA binding, and activity of Smads in cervical cancer cell lines treated with TGF-β1 (p<0.01). CONCLUSION:EGCG and green tea extract suppressed TGF-β-induced EMT in Hela and SiHa cells, and the underlying molecular mechanism may be related to the ROS generation and Smad signaling pathway.
Authors: Sze Wan Hung; Yiran Li; Xiaoyan Chen; Kai On Chu; Yiwei Zhao; Yingyu Liu; Xi Guo; Gene Chi-Wai Man; Chi Chiu Wang Journal: Front Pharmacol Date: 2022-07-04 Impact factor: 5.988