Molecular Target Identification of Gossypol Against Cervical Cancer Based on Target Fishing Technology

<b>Objectives</b>: This study aims to investigate the impact of Gossypol on human cervical cancer cells and elucidate its mechanism of action to establish a foundation for further clinical investigations. <b>Methods</b>: Cell proliferation, migration, and invasion were evalua...

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Main Authors: Jinyan Li, Rayisa Asat, Wenying Li, Parwen Parhat, Yue Ma, Yinglan Ma, Min Li
Format: Article
Language:English
Published: MDPI AG 2025-06-01
Series:Pharmaceutics
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Online Access:https://www.mdpi.com/1999-4923/17/7/861
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author Jinyan Li
Rayisa Asat
Wenying Li
Parwen Parhat
Yue Ma
Yinglan Ma
Min Li
author_facet Jinyan Li
Rayisa Asat
Wenying Li
Parwen Parhat
Yue Ma
Yinglan Ma
Min Li
author_sort Jinyan Li
collection DOAJ
description <b>Objectives</b>: This study aims to investigate the impact of Gossypol on human cervical cancer cells and elucidate its mechanism of action to establish a foundation for further clinical investigations. <b>Methods</b>: Cell proliferation, migration, and invasion were evaluated through CCK−8, wound healing, and Transwell assays. Fe<sub>3</sub>O<sub>4</sub>-BP-Gossypol (Fe<sub>3</sub>O<sub>4</sub>@Gossypol) conjugates were synthesized by linking Fe<sub>3</sub>O<sub>4</sub> with Gossypol using benzophenone crosslinking. Successful conjugation was confirmed through scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), and ultraviolet–visible spectrophotometry (UV-Vis). Subsequent to co-incubation with HeLa cell lysates, Fe<sub>3</sub>O<sub>4</sub>@Gossypol complexes facilitated the magnetic enrichment and purification of target proteins, which were identified using high-resolution mass spectrometry (HR-MS). The identified targets underwent KEGG pathway and GO analyses, followed by molecular docking with Gossypol. HeLa cells were exposed to Gossypol at concentrations of 7.48, 14.96, and 29.92 μmol·L<sup>−1</sup> for 48 h, and protein expression levels were quantified via Western blotting. <b>Results</b>: Gossypol notably suppressed cervical cancer cell proliferation, migration, and invasion. The integration of target fishing, network pharmacology, and molecular docking highlighted PIK3R2, MAPK1, and GRB2 as potential therapeutic targets. Western blot analysis revealed a dose-dependent reduction in PIK3R2, GRB2, and MAPK1 expression in Gossypol-treated groups compared to controls (<i>p</i> < 0.05). <b>Conclusions</b>: Gossypol may exhibit anti-cervical cancer effects by modulating the PI3K/AKT signaling pathway.
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spelling doaj-art-6a2d95c1ded4459682ec445827e41b6f2025-08-20T03:32:27ZengMDPI AGPharmaceutics1999-49232025-06-0117786110.3390/pharmaceutics17070861Molecular Target Identification of Gossypol Against Cervical Cancer Based on Target Fishing TechnologyJinyan Li0Rayisa Asat1Wenying Li2Parwen Parhat3Yue Ma4Yinglan Ma5Min Li6College of Pharmacy, Xinjiang Medical University, Urumqi 830011, ChinaCollege of Pharmacy, Xinjiang Medical University, Urumqi 830011, ChinaCollege of Pharmacy, Xinjiang Medical University, Urumqi 830011, ChinaCollege of Pharmacy, Xinjiang Medical University, Urumqi 830011, ChinaCollege of Pharmacy, Xinjiang Medical University, Urumqi 830011, ChinaCollege of Pharmacy, Xinjiang Medical University, Urumqi 830011, ChinaCollege of Pharmacy, Xinjiang Medical University, Urumqi 830011, China<b>Objectives</b>: This study aims to investigate the impact of Gossypol on human cervical cancer cells and elucidate its mechanism of action to establish a foundation for further clinical investigations. <b>Methods</b>: Cell proliferation, migration, and invasion were evaluated through CCK−8, wound healing, and Transwell assays. Fe<sub>3</sub>O<sub>4</sub>-BP-Gossypol (Fe<sub>3</sub>O<sub>4</sub>@Gossypol) conjugates were synthesized by linking Fe<sub>3</sub>O<sub>4</sub> with Gossypol using benzophenone crosslinking. Successful conjugation was confirmed through scanning electron microscopy (SEM), Fourier transform infrared spectroscopy (FT-IR), and ultraviolet–visible spectrophotometry (UV-Vis). Subsequent to co-incubation with HeLa cell lysates, Fe<sub>3</sub>O<sub>4</sub>@Gossypol complexes facilitated the magnetic enrichment and purification of target proteins, which were identified using high-resolution mass spectrometry (HR-MS). The identified targets underwent KEGG pathway and GO analyses, followed by molecular docking with Gossypol. HeLa cells were exposed to Gossypol at concentrations of 7.48, 14.96, and 29.92 μmol·L<sup>−1</sup> for 48 h, and protein expression levels were quantified via Western blotting. <b>Results</b>: Gossypol notably suppressed cervical cancer cell proliferation, migration, and invasion. The integration of target fishing, network pharmacology, and molecular docking highlighted PIK3R2, MAPK1, and GRB2 as potential therapeutic targets. Western blot analysis revealed a dose-dependent reduction in PIK3R2, GRB2, and MAPK1 expression in Gossypol-treated groups compared to controls (<i>p</i> < 0.05). <b>Conclusions</b>: Gossypol may exhibit anti-cervical cancer effects by modulating the PI3K/AKT signaling pathway.https://www.mdpi.com/1999-4923/17/7/861gossypoltarget fishingcervical cancermagnetic microspheres
spellingShingle Jinyan Li
Rayisa Asat
Wenying Li
Parwen Parhat
Yue Ma
Yinglan Ma
Min Li
Molecular Target Identification of Gossypol Against Cervical Cancer Based on Target Fishing Technology
Pharmaceutics
gossypol
target fishing
cervical cancer
magnetic microspheres
title Molecular Target Identification of Gossypol Against Cervical Cancer Based on Target Fishing Technology
title_full Molecular Target Identification of Gossypol Against Cervical Cancer Based on Target Fishing Technology
title_fullStr Molecular Target Identification of Gossypol Against Cervical Cancer Based on Target Fishing Technology
title_full_unstemmed Molecular Target Identification of Gossypol Against Cervical Cancer Based on Target Fishing Technology
title_short Molecular Target Identification of Gossypol Against Cervical Cancer Based on Target Fishing Technology
title_sort molecular target identification of gossypol against cervical cancer based on target fishing technology
topic gossypol
target fishing
cervical cancer
magnetic microspheres
url https://www.mdpi.com/1999-4923/17/7/861
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