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      Stigmasterol Simultaneously Induces Apoptosis and Protective Autophagy by Inhibiting Akt/mTOR Pathway in Gastric Cancer Cells

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          Abstract

          Background

          Stigmasterol (SS) has been proven to possess potential anticancer activities in several cancer cell lines, but its molecular mechanism is still unknown. Thus, we investigated whether SS has the capabilities of inducing autophagy and its molecular mechanisms in gastric cancer cells.

          Methods

          We used CCK8 assay, clone formation assay, and EdU proliferation assay to assess the effects of SS on cell proliferation in SGC-7901 and MGC-803 cells in vitro, and its inhibition on the tumor growth of gastric cancer was observed in vivo. Apoptosis induced by SS was demonstrated using Hoechst and TUNEL staining, annexin V-FITC/PI assay. Immunofluorescence staining is used to detect the formation of autophagosomes triggered by SS. Apoptosis and autophagy related proteins were analyzed by western blot.

          Results

          The results indicated that SS treatment inhibited cell proliferation in SGC-7901 and MGC-803 cells. Furthermore, SS treatment induced apoptosis and autophagy by blocking Akt/mTOR signaling pathway. The pretreatment with the Akt inhibitor MK-2206 could promote apoptosis and autophagy induced by SS, predicting that Akt/mTOR pathway is involved in SS-induced apoptosis and autophagy. In addition, blockade of autophagy with 3-MA (an inhibitor of autophagy) enhanced SS-induced apoptosis in SGC-7901 and MGC-803 cells, implying that autophagy mediated by SS plays a cytoprotective role against apoptosis. Finally, an in vivo study demonstrated that tumor growth of gastric cancer was suppressed by SS in a xenograft model.

          Conclusion

          Our findings illustrate for the first time that SS simultaneously induces apoptosis and protective autophagy by inhibiting Akt/mTOR pathway in gastric cancer cells, and SS may become a potential anticancer drug in treating gastric cancer in the future.

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          Most cited references42

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          Guidelines for the use and interpretation of assays for monitoring autophagy (3rd edition).

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            Methods in mammalian autophagy research.

            Autophagy has been implicated in many physiological and pathological processes. Accordingly, there is a growing scientific need to accurately identify, quantify, and manipulate the process of autophagy. However, as autophagy involves dynamic and complicated processes, it is often analyzed incorrectly. In this Primer, we discuss methods to monitor autophagy and to modulate autophagic activity, with a primary focus on mammalian macroautophagy. 2010 Elsevier Inc. All rights reserved.
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              Self-eating and self-killing: crosstalk between autophagy and apoptosis.

              The functional relationship between apoptosis ('self-killing') and autophagy ('self-eating') is complex in the sense that, under certain circumstances, autophagy constitutes a stress adaptation that avoids cell death (and suppresses apoptosis), whereas in other cellular settings, it constitutes an alternative cell-death pathway. Autophagy and apoptosis may be triggered by common upstream signals, and sometimes this results in combined autophagy and apoptosis; in other instances, the cell switches between the two responses in a mutually exclusive manner. On a molecular level, this means that the apoptotic and autophagic response machineries share common pathways that either link or polarize the cellular responses.
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                Author and article information

                Contributors
                Journal
                Front Oncol
                Front Oncol
                Front. Oncol.
                Frontiers in Oncology
                Frontiers Media S.A.
                2234-943X
                23 February 2021
                2021
                : 11
                : 629008
                Affiliations
                [1] 1 Key Laboratory of Tropical Translational Medicine of the Ministry of Education & Hainan Provincial Key Laboratory of Tropical Medicine, Hainan Medical University , Haikou, China
                [2] 2 Schools of Basic Medicine and Life Sciences, Hainan Medical University , Haikou, China
                Author notes

                Edited by: Andrew Zloza, Rush University Medical Center, United States

                Reviewed by: XiuJuan Qu, The First Affiliated Hospital of China Medical University, China; Amanda Marzo, Rush University, United States; Hui-Min David Wang, National Chung Hsing University, Taiwan

                *Correspondence: Songlin Zhou, zhousonglin106@ 123456163.com

                This article was submitted to Cancer Molecular Targets and Therapeutics, a section of the journal Frontiers in Oncology

                Article
                10.3389/fonc.2021.629008
                7940753
                33708631
                10e59f68-68ca-4d4b-abf6-735b8b57dbb9
                Copyright © 2021 Zhao, Zhang, Wang, Lin and Zhou

                This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms.

                History
                : 13 November 2020
                : 05 January 2021
                Page count
                Figures: 7, Tables: 0, Equations: 0, References: 42, Pages: 11, Words: 4772
                Funding
                Funded by: Key Research and Development Project of Hainan Province 10.13039/501100013142
                Award ID: ZDYF2019177
                Categories
                Oncology
                Original Research

                Oncology & Radiotherapy
                stigmasterol,apoptosis,autophagy,akt/mtor signaling pathway,gastric cancer cells

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