基于网络药理学和体外实验的牡蛎糖原抗甲状腺乳头状癌作用及机制研究

Anti-tumor effects and potential mechanisms of oyster glycogen in papillary thyroid carcinoma: a network pharmacology and in vitro experimental study

摘要:

背景与目的 甲状腺乳头状癌（PTC）是最常见的甲状腺恶性肿瘤，部分患者仍面临复发、转移及放射性碘难治等问题。牡蛎糖原具有抗肿瘤活性，但其对PTC的作用及机制尚不明确。本研究结合网络药理学分析、分子对接及体外试验，探讨牡蛎糖原抗PTC的潜在作用机制。方法 通过ChEMBL、ETCM及SwissTargetPrediction数据库预测牡蛎糖原潜在作用靶点，通过GeneCards、OMIM及PharmGKB数据库筛选PTC相关靶点，获得共同靶点后进行蛋白质相互作用（PPI）网络构建、GO功能富集及KEGG通路分析，并筛选核心靶点。采用AutoDock Vina软件进行分子对接验证。以TPC-1细胞为研究对象，采用CCK-8法检测不同浓度牡蛎糖原对细胞活力的影响，采用Annexin V-FITC/PI双染流式细胞术检测细胞凋亡情况。结果 共筛得牡蛎糖原与PTC共同靶点48个。PPI网络分析筛选出PTGS2、GSK3β、AR及PGR等核心靶点。GO和KEGG分析显示，共同靶点主要涉及上皮细胞增殖调控、转录因子结合及PI3K-Akt信号通路等生物学过程。分子对接结果显示，牡蛎糖原与PTGS2、GSK3β、AR及PGR均具有较好的结合活性，其中与PTGS2结合能力最强（-9.1 kcal/mol）。CCK-8实验显示，牡蛎糖原对TPC-1细胞的活性抑制具有剂量依赖性和时间依赖性，48 h时IC₂₀和IC₅₀分别为37.7 μmol/L和233.6 μmol/L。流式细胞术结果显示，经37.7 μmol/L牡蛎糖原处理48 h后，TPC-1细胞总凋亡率由0.20%升高至18.61%（P<0.01）。结论 牡蛎糖原能够抑制PTC细胞增殖并促进细胞凋亡，其作用可能与PTGS2、GSK3β及PI3K-Akt信号通路相关。上述机制仍需进一步通过分子生物学实验进行验证。

Abstract:

Background and Aims Papillary thyroid carcinoma (PTC) is the most common type of thyroid malignancy. Despite favorable outcomes in most patients, recurrence, metastasis, and radioiodine-refractory disease remain important clinical challenges. Glycogen, a major bioactive component derived from oyster, has demonstrated antitumor activity; however, its effects and underlying mechanisms in PTC remain unclear. This study aimed to investigate the potential anti-PTC mechanisms of oyster glycogen through network pharmacology, molecular docking, and in vitro experiments.Methods Potential targets of oyster glycogen were predicted using the ChEMBL, ETCM, and SwissTargetPrediction databases, while PTC-related targets were collected from the GeneCards, OMIM, and PharmGKB databases. Common targets were identified and subjected to protein-protein interaction (PPI) network construction, Gene Ontology (GO), and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses. Core targets were screened and further validated by molecular docking using AutoDock Vina. TPC-1 cells were treated with different concentrations of oyster glycogen. Cell viability was evaluated using the CCK-8 assay, and apoptosis was assessed by Annexin V-FITC/PI staining followed by flow cytometry.Results A total of 48 common targets were identified between oyster glycogen and PTC. PPI analysis revealed PTGS2, GSK3β, AR, and PGR as key targets. GO and KEGG analyses suggested that these targets were mainly involved in epithelial cell proliferation, transcription factor binding, and the PI3K-Akt signaling pathway. Molecular docking demonstrated favorable binding affinities between oyster glycogen and the core targets, with the strongest interaction observed for PTGS2 (-9.1 kcal/mol). CCK-8 assays showed that oyster glycogen inhibited TPC-1 cell viability in a dose- and time-dependent manner, with IC₂₀ and IC₅₀ values of 37.7 μmol/L and 233.6 μmol/L, respectively, at 48 h. Flow cytometric analysis demonstrated that treatment with 37.7 μmol/L oyster glycogen for 48 h increased the total apoptosis rate from 0.20% to 18.61% (P<0.01).Conclusion Oyster glycogen inhibits the proliferation of PTC cells and promotes apoptosis. These effects may be associated with PTGS2, GSK3β, and the PI3K-Akt signaling pathway. Further molecular studies are required to validate the underlying mechanisms.