Antiproliferative Activity and Apoptotic Mechanisms of β-Sitosterol and Its Derivatives as Anti-Breast Cancer Agents: In Silico and In Vitro

Mutakin Mutakin,1 Lauren Pangestu,2 Nafisa Nurfatia Hidayat,2 Fajar Fauzi Abdullah,3 Yuni Elsa Hadisaputri2 1Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Padjadjaran, West Java, Indonesia; 2Department of Pharmaceutical Biology, Faculty of Pharmacy,...

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Main Authors: Mutakin M, Pangestu L, Hidayat NN, Abdullah FF, Hadisaputri YE
Format: Article
Language:English
Published: Dove Medical Press 2025-02-01
Series:Journal of Experimental Pharmacology
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Online Access:https://www.dovepress.com/antiproliferative-activity-and-apoptotic-mechanisms-of--sitosterol-and-peer-reviewed-fulltext-article-JEP
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author Mutakin M
Pangestu L
Hidayat NN
Abdullah FF
Hadisaputri YE
author_facet Mutakin M
Pangestu L
Hidayat NN
Abdullah FF
Hadisaputri YE
author_sort Mutakin M
collection DOAJ
description Mutakin Mutakin,1 Lauren Pangestu,2 Nafisa Nurfatia Hidayat,2 Fajar Fauzi Abdullah,3 Yuni Elsa Hadisaputri2 1Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Padjadjaran, West Java, Indonesia; 2Department of Pharmaceutical Biology, Faculty of Pharmacy, Universitas Padjadjaran, West Java, Indonesia; 3Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Garut, West Java, IndonesiaCorrespondence: Mutakin Mutakin, Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Padjadjaran, Jl. Raya Bandung-Sumedang KM.21, Jatinangor, West Java, 45363, Indonesia, Tel +62-22-84288888 Ext. 3510, Email mutakin@unpad.ac.idIntroduction: Breast cancer has become the most frequently diagnosed cancer worldwide. Beta-sitosterol and its derivatives have been explored for its anticancer properties. Therefore, this study aims to analyze the testing procedure carried out on MCF7 and MDA-MB-231 breast cancer cells, as well as MCF 10A non-cancerous breast epithelial cells.Methods: The compounds tested included β-sitosterol and its derivatives: 3β-galactose sitosterol, sitostenone, 3β-glucose sitosterol, poriferasta-5, 22E, 25-trien-3β-ol, and 22-dehydrocholesterol. Cytotoxicity assay was conducted using the PrestoBlue™ Cell Viability Reagent on MCF-7, MDA-MB-231, and MCF 10A cells. The compounds with the highest and lowest cytotoxicity were further analyzed for their mechanisms of action through cell morphology assessments and molecular docking studies. mRNA expression levels were also evaluated to confirm the findings.Results: The results showed that 3β-glucose sitosterol exhibited the most promising cytotoxic activity with IC50 values against MCF7, MDA-MB-231 breast cancer cells, and MCF 10A non-cancerous cells of 265 μg/mL, 393.862 μg/mL, and 806.833 μg/mL, respectively. Molecular docking simulations showed that the compound is bound to estrogen receptor beta and caspase-3, suggesting a potential mechanism of action as evidenced by the best binding energy of − 6.94 kcal/mol and inhibition constant values of 8.16 μM. Furthermore, gene expression analysis confirmed the induction of apoptosis through the upregulation of caspase-9 and caspase-3 mRNA expression.Conclusion: Based on the results, β-sitosterol and its derivatives, particularly 3β-glucose sitosterol, show as the most promising potential adjuvant therapy for hormone-positive breast cancer. Keywords: β-sitosterol, 3β-glucose sitosterol, breast cancer, MCF7, MDA-MB-231, ESR2
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spelling doaj-art-5fc4083cc60843e5b64d8cd28442d31d2025-08-20T02:48:42ZengDove Medical PressJournal of Experimental Pharmacology1179-14542025-02-01Volume 17107121100226Antiproliferative Activity and Apoptotic Mechanisms of β-Sitosterol and Its Derivatives as Anti-Breast Cancer Agents: In Silico and In VitroMutakin MPangestu LHidayat NNAbdullah FFHadisaputri YEMutakin Mutakin,1 Lauren Pangestu,2 Nafisa Nurfatia Hidayat,2 Fajar Fauzi Abdullah,3 Yuni Elsa Hadisaputri2 1Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Padjadjaran, West Java, Indonesia; 2Department of Pharmaceutical Biology, Faculty of Pharmacy, Universitas Padjadjaran, West Java, Indonesia; 3Department of Chemistry, Faculty of Mathematics and Natural Science, Universitas Garut, West Java, IndonesiaCorrespondence: Mutakin Mutakin, Department of Pharmaceutical Analysis and Medicinal Chemistry, Faculty of Pharmacy, Universitas Padjadjaran, Jl. Raya Bandung-Sumedang KM.21, Jatinangor, West Java, 45363, Indonesia, Tel +62-22-84288888 Ext. 3510, Email mutakin@unpad.ac.idIntroduction: Breast cancer has become the most frequently diagnosed cancer worldwide. Beta-sitosterol and its derivatives have been explored for its anticancer properties. Therefore, this study aims to analyze the testing procedure carried out on MCF7 and MDA-MB-231 breast cancer cells, as well as MCF 10A non-cancerous breast epithelial cells.Methods: The compounds tested included β-sitosterol and its derivatives: 3β-galactose sitosterol, sitostenone, 3β-glucose sitosterol, poriferasta-5, 22E, 25-trien-3β-ol, and 22-dehydrocholesterol. Cytotoxicity assay was conducted using the PrestoBlue™ Cell Viability Reagent on MCF-7, MDA-MB-231, and MCF 10A cells. The compounds with the highest and lowest cytotoxicity were further analyzed for their mechanisms of action through cell morphology assessments and molecular docking studies. mRNA expression levels were also evaluated to confirm the findings.Results: The results showed that 3β-glucose sitosterol exhibited the most promising cytotoxic activity with IC50 values against MCF7, MDA-MB-231 breast cancer cells, and MCF 10A non-cancerous cells of 265 μg/mL, 393.862 μg/mL, and 806.833 μg/mL, respectively. Molecular docking simulations showed that the compound is bound to estrogen receptor beta and caspase-3, suggesting a potential mechanism of action as evidenced by the best binding energy of − 6.94 kcal/mol and inhibition constant values of 8.16 μM. Furthermore, gene expression analysis confirmed the induction of apoptosis through the upregulation of caspase-9 and caspase-3 mRNA expression.Conclusion: Based on the results, β-sitosterol and its derivatives, particularly 3β-glucose sitosterol, show as the most promising potential adjuvant therapy for hormone-positive breast cancer. Keywords: β-sitosterol, 3β-glucose sitosterol, breast cancer, MCF7, MDA-MB-231, ESR2https://www.dovepress.com/antiproliferative-activity-and-apoptotic-mechanisms-of--sitosterol-and-peer-reviewed-fulltext-article-JEPβ-sitosterol3β-glucose sitosterolbreast cancermcf7mda-mb-231esr2
spellingShingle Mutakin M
Pangestu L
Hidayat NN
Abdullah FF
Hadisaputri YE
Antiproliferative Activity and Apoptotic Mechanisms of β-Sitosterol and Its Derivatives as Anti-Breast Cancer Agents: In Silico and In Vitro
Journal of Experimental Pharmacology
β-sitosterol
3β-glucose sitosterol
breast cancer
mcf7
mda-mb-231
esr2
title Antiproliferative Activity and Apoptotic Mechanisms of β-Sitosterol and Its Derivatives as Anti-Breast Cancer Agents: In Silico and In Vitro
title_full Antiproliferative Activity and Apoptotic Mechanisms of β-Sitosterol and Its Derivatives as Anti-Breast Cancer Agents: In Silico and In Vitro
title_fullStr Antiproliferative Activity and Apoptotic Mechanisms of β-Sitosterol and Its Derivatives as Anti-Breast Cancer Agents: In Silico and In Vitro
title_full_unstemmed Antiproliferative Activity and Apoptotic Mechanisms of β-Sitosterol and Its Derivatives as Anti-Breast Cancer Agents: In Silico and In Vitro
title_short Antiproliferative Activity and Apoptotic Mechanisms of β-Sitosterol and Its Derivatives as Anti-Breast Cancer Agents: In Silico and In Vitro
title_sort antiproliferative activity and apoptotic mechanisms of beta sitosterol and its derivatives as anti breast cancer agents in silico and in vitro
topic β-sitosterol
3β-glucose sitosterol
breast cancer
mcf7
mda-mb-231
esr2
url https://www.dovepress.com/antiproliferative-activity-and-apoptotic-mechanisms-of--sitosterol-and-peer-reviewed-fulltext-article-JEP
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