Global Identification of Anti-Melanoma Cellular Targets by Photochemically Induced Coupling of <i>L-Shikonin</i> Reactions on the Surface of Magnetic Particles

<b>Background:</b> <i>L-Shikonin</i>, an active component of Arnebia euchroma (Royle) Johnst., has remarkable pharmacological effects, particularly in its anti-tumour activity. Nonetheless, the specific targets and mechanisms of action remain to be further explored. <b>...

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Main Authors: Min Li, Wenying Li, Fang Xu, Yiping Pu, Jianguang Li
Format: Article
Language:English
Published: MDPI AG 2024-12-01
Series:Pharmaceutics
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Online Access:https://www.mdpi.com/1999-4923/16/12/1543
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author Min Li
Wenying Li
Fang Xu
Yiping Pu
Jianguang Li
author_facet Min Li
Wenying Li
Fang Xu
Yiping Pu
Jianguang Li
author_sort Min Li
collection DOAJ
description <b>Background:</b> <i>L-Shikonin</i>, an active component of Arnebia euchroma (Royle) Johnst., has remarkable pharmacological effects, particularly in its anti-tumour activity. Nonetheless, the specific targets and mechanisms of action remain to be further explored. <b>Methods:</b> A novel Fe<sub>3</sub>O<sub>4</sub>@<i>L-Shikonin</i> was designed and synthesized in this study by linking Fe<sub>3</sub>O<sub>4</sub> and <i>L-Shikonin</i> through benzophenone. Fe<sub>3</sub>O<sub>4</sub>@<i>L-Shikonin</i> was characterized using several techniques, including scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), and drug removal methods, to determine the content of <i>L-Shikonin</i> on the surface of the magnetic particles. Target hooking technology was utilized to identify the target proteins of the compound in melanoma. The synthesized Fe<sub>3</sub>O<sub>4</sub>@<i>L-Shikonin</i> was co-incubated with A375 cell lysate, followed by the target proteins, which were purified by magnetic enrichment using magnetic microspheres and identified by high-resolution mass spectrometry. <b>Results:</b> AutoDock Vina software was employed for molecular docking analysis, where it was found that <i>L-Shikonin</i> targets RPN1, CPEB4, and HNRNPUL1 proteins. Subsequently, A375 cells were treated with <i>L-Shikonin</i> at different concentrations (2.5, 5.0, 10.0 μM) for 48 h, and the expressions of the three proteins were observed. The results showed a significant reduction in the relative expression of CPEB4 in the high-dose group compared to the control group (<i>p</i> < 0.01). Moreover, the relative expression of HNRNPUL1 was decreased in the medium- and high-dose groups (<i>p</i> < 0.05). <b>Conclusions:</b> This study initially revealed from the source that <i>L-Shikonin</i> may regulate melanoma-specific markers, melanosomes, tyrosine kinases related to abnormal tyrosine metabolism, and melanoma through multiple targets such as CPEB4 and HNRNPUL1. Proliferation and metastasis work together to exert an anti-melanoma mechanism, which provides a new idea for the follow-up study of the molecular pharmacological mechanism of the complex system of total naphthoquinones in <i>Arnebia euchroma</i> (Royle) Johns.
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spelling doaj-art-b34449d41f944c4381f2ffe2c336cd752024-12-27T14:46:26ZengMDPI AGPharmaceutics1999-49232024-12-011612154310.3390/pharmaceutics16121543Global Identification of Anti-Melanoma Cellular Targets by Photochemically Induced Coupling of <i>L-Shikonin</i> Reactions on the Surface of Magnetic ParticlesMin Li0Wenying Li1Fang Xu2Yiping Pu3Jianguang Li4College 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 Second Medical College, Karamay 834000, China<b>Background:</b> <i>L-Shikonin</i>, an active component of Arnebia euchroma (Royle) Johnst., has remarkable pharmacological effects, particularly in its anti-tumour activity. Nonetheless, the specific targets and mechanisms of action remain to be further explored. <b>Methods:</b> A novel Fe<sub>3</sub>O<sub>4</sub>@<i>L-Shikonin</i> was designed and synthesized in this study by linking Fe<sub>3</sub>O<sub>4</sub> and <i>L-Shikonin</i> through benzophenone. Fe<sub>3</sub>O<sub>4</sub>@<i>L-Shikonin</i> was characterized using several techniques, including scanning electron microscopy (SEM), Fourier-transform infrared spectroscopy (FT-IR), and drug removal methods, to determine the content of <i>L-Shikonin</i> on the surface of the magnetic particles. Target hooking technology was utilized to identify the target proteins of the compound in melanoma. The synthesized Fe<sub>3</sub>O<sub>4</sub>@<i>L-Shikonin</i> was co-incubated with A375 cell lysate, followed by the target proteins, which were purified by magnetic enrichment using magnetic microspheres and identified by high-resolution mass spectrometry. <b>Results:</b> AutoDock Vina software was employed for molecular docking analysis, where it was found that <i>L-Shikonin</i> targets RPN1, CPEB4, and HNRNPUL1 proteins. Subsequently, A375 cells were treated with <i>L-Shikonin</i> at different concentrations (2.5, 5.0, 10.0 μM) for 48 h, and the expressions of the three proteins were observed. The results showed a significant reduction in the relative expression of CPEB4 in the high-dose group compared to the control group (<i>p</i> < 0.01). Moreover, the relative expression of HNRNPUL1 was decreased in the medium- and high-dose groups (<i>p</i> < 0.05). <b>Conclusions:</b> This study initially revealed from the source that <i>L-Shikonin</i> may regulate melanoma-specific markers, melanosomes, tyrosine kinases related to abnormal tyrosine metabolism, and melanoma through multiple targets such as CPEB4 and HNRNPUL1. Proliferation and metastasis work together to exert an anti-melanoma mechanism, which provides a new idea for the follow-up study of the molecular pharmacological mechanism of the complex system of total naphthoquinones in <i>Arnebia euchroma</i> (Royle) Johns.https://www.mdpi.com/1999-4923/16/12/1543<i>L-Shikonin</i>surface graftingcoupling molecules in medicinetarget fishing hookmagnetic nanoparticlemelanoma
spellingShingle Min Li
Wenying Li
Fang Xu
Yiping Pu
Jianguang Li
Global Identification of Anti-Melanoma Cellular Targets by Photochemically Induced Coupling of <i>L-Shikonin</i> Reactions on the Surface of Magnetic Particles
Pharmaceutics
<i>L-Shikonin</i>
surface grafting
coupling molecules in medicine
target fishing hook
magnetic nanoparticle
melanoma
title Global Identification of Anti-Melanoma Cellular Targets by Photochemically Induced Coupling of <i>L-Shikonin</i> Reactions on the Surface of Magnetic Particles
title_full Global Identification of Anti-Melanoma Cellular Targets by Photochemically Induced Coupling of <i>L-Shikonin</i> Reactions on the Surface of Magnetic Particles
title_fullStr Global Identification of Anti-Melanoma Cellular Targets by Photochemically Induced Coupling of <i>L-Shikonin</i> Reactions on the Surface of Magnetic Particles
title_full_unstemmed Global Identification of Anti-Melanoma Cellular Targets by Photochemically Induced Coupling of <i>L-Shikonin</i> Reactions on the Surface of Magnetic Particles
title_short Global Identification of Anti-Melanoma Cellular Targets by Photochemically Induced Coupling of <i>L-Shikonin</i> Reactions on the Surface of Magnetic Particles
title_sort global identification of anti melanoma cellular targets by photochemically induced coupling of i l shikonin i reactions on the surface of magnetic particles
topic <i>L-Shikonin</i>
surface grafting
coupling molecules in medicine
target fishing hook
magnetic nanoparticle
melanoma
url https://www.mdpi.com/1999-4923/16/12/1543
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