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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Summary:<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.
ISSN:1999-4923