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Inhibition of TMEM16A by Natural Product Silibinin: Potential Lead Compounds for Treatment of Lung Adenocarcinoma

Background: Effective anticancer therapy can be achieved by identifying novel tumor-specific drug targets and screening of new drugs. Recently, TMEM16A has been identified to be overexpressed in lung adenocarcinoma, and inhibitors of TMEM16A showed obvious antitumor efficacy. Methods: YFP fluorescen...

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Autores principales: Guo, Shuai, Bai, Xue, Liu, Yufei, Shi, Sai, Wang, Xuzhao, Zhan, Yong, Kang, Xianjiang, Chen, Yafei, An, Hailong
Formato: Online Artículo Texto
Lenguaje:English
Publicado: Frontiers Media S.A. 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079988/
https://www.ncbi.nlm.nih.gov/pubmed/33935737
http://dx.doi.org/10.3389/fphar.2021.643489
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author Guo, Shuai
Bai, Xue
Liu, Yufei
Shi, Sai
Wang, Xuzhao
Zhan, Yong
Kang, Xianjiang
Chen, Yafei
An, Hailong
author_facet Guo, Shuai
Bai, Xue
Liu, Yufei
Shi, Sai
Wang, Xuzhao
Zhan, Yong
Kang, Xianjiang
Chen, Yafei
An, Hailong
author_sort Guo, Shuai
collection PubMed
description Background: Effective anticancer therapy can be achieved by identifying novel tumor-specific drug targets and screening of new drugs. Recently, TMEM16A has been identified to be overexpressed in lung adenocarcinoma, and inhibitors of TMEM16A showed obvious antitumor efficacy. Methods: YFP fluorescence quenching and whole-cell patch clamp experiments were used to explore the inhibitory effect of silibinin on TMEM16A. Molecular docking and site-directed mutagenesis were performed to confirm the binding sites of silibinin and TMEM16A. MTT assay, wound healing assay, and annexin-V assay were used to detect the effect of silibinin on cancer cell proliferation, migration, and apoptosis. shRNA was transfected into LA795 cells to knock down the expression of endogenous TMEM16A. Tumor xenograft mice combined with Western blot experiments reveal the inhibitory effect and mechanism of silibinin in vivo. Results: Silibinin concentration dependently inhibited the whole-cell current of TMEM16A with an IC(50) of 30.90 ± 2.10 μM. The putative binding sites of silibinin in TMEM16A were K384, R515, and R535. The proliferation and migration of LA795 cells were downregulated by silibinin, and the inhibition effect can be abolished by knockdown of the endogenous TMEM16A. Further, silibinin was injected to tumor xenograft mice which exhibited significant antitumor activity without weight loss. Finally, Western blotting results showed the mechanism of silibinin inhibiting lung adenocarcinoma was through apoptosis and downregulation of cyclin D1. Conclusion: Silibinin is a novel TMEM16A inhibitor, and it can be used as a lead compound for the development of lung adenocarcinoma therapy drugs.
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spelling pubmed-80799882021-04-29 Inhibition of TMEM16A by Natural Product Silibinin: Potential Lead Compounds for Treatment of Lung Adenocarcinoma Guo, Shuai Bai, Xue Liu, Yufei Shi, Sai Wang, Xuzhao Zhan, Yong Kang, Xianjiang Chen, Yafei An, Hailong Front Pharmacol Pharmacology Background: Effective anticancer therapy can be achieved by identifying novel tumor-specific drug targets and screening of new drugs. Recently, TMEM16A has been identified to be overexpressed in lung adenocarcinoma, and inhibitors of TMEM16A showed obvious antitumor efficacy. Methods: YFP fluorescence quenching and whole-cell patch clamp experiments were used to explore the inhibitory effect of silibinin on TMEM16A. Molecular docking and site-directed mutagenesis were performed to confirm the binding sites of silibinin and TMEM16A. MTT assay, wound healing assay, and annexin-V assay were used to detect the effect of silibinin on cancer cell proliferation, migration, and apoptosis. shRNA was transfected into LA795 cells to knock down the expression of endogenous TMEM16A. Tumor xenograft mice combined with Western blot experiments reveal the inhibitory effect and mechanism of silibinin in vivo. Results: Silibinin concentration dependently inhibited the whole-cell current of TMEM16A with an IC(50) of 30.90 ± 2.10 μM. The putative binding sites of silibinin in TMEM16A were K384, R515, and R535. The proliferation and migration of LA795 cells were downregulated by silibinin, and the inhibition effect can be abolished by knockdown of the endogenous TMEM16A. Further, silibinin was injected to tumor xenograft mice which exhibited significant antitumor activity without weight loss. Finally, Western blotting results showed the mechanism of silibinin inhibiting lung adenocarcinoma was through apoptosis and downregulation of cyclin D1. Conclusion: Silibinin is a novel TMEM16A inhibitor, and it can be used as a lead compound for the development of lung adenocarcinoma therapy drugs. Frontiers Media S.A. 2021-04-14 /pmc/articles/PMC8079988/ /pubmed/33935737 http://dx.doi.org/10.3389/fphar.2021.643489 Text en Copyright © 2021 Guo, Bai, Liu, Shi, Wang, Zhan, Kang, Chen and An. https://creativecommons.org/licenses/by/4.0/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.
spellingShingle Pharmacology
Guo, Shuai
Bai, Xue
Liu, Yufei
Shi, Sai
Wang, Xuzhao
Zhan, Yong
Kang, Xianjiang
Chen, Yafei
An, Hailong
Inhibition of TMEM16A by Natural Product Silibinin: Potential Lead Compounds for Treatment of Lung Adenocarcinoma
title Inhibition of TMEM16A by Natural Product Silibinin: Potential Lead Compounds for Treatment of Lung Adenocarcinoma
title_full Inhibition of TMEM16A by Natural Product Silibinin: Potential Lead Compounds for Treatment of Lung Adenocarcinoma
title_fullStr Inhibition of TMEM16A by Natural Product Silibinin: Potential Lead Compounds for Treatment of Lung Adenocarcinoma
title_full_unstemmed Inhibition of TMEM16A by Natural Product Silibinin: Potential Lead Compounds for Treatment of Lung Adenocarcinoma
title_short Inhibition of TMEM16A by Natural Product Silibinin: Potential Lead Compounds for Treatment of Lung Adenocarcinoma
title_sort inhibition of tmem16a by natural product silibinin: potential lead compounds for treatment of lung adenocarcinoma
topic Pharmacology
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8079988/
https://www.ncbi.nlm.nih.gov/pubmed/33935737
http://dx.doi.org/10.3389/fphar.2021.643489
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