目的 本研究旨在基于机器学习技术筛选半乳糖凝集素-3（Gal-3）的小分子抑制剂,并探讨其与Gal-3的结合模式及对肝内胆管癌（intrahepatic cholangiocarcinoma,ICC）细胞增殖和迁移的影响。方法 通过生物信息学和免疫组化分析Gal-3在ICC组织中的表达及其与预后的关系;使用Schrödinger软件对Gal-3结构和TargetMol L4000化合物库进行处理,采用分子对接技术进行虚拟筛选,结合聚类分析和人工筛选得到候选抑制剂;不同浓度抑制剂作用ICC细胞系24 h和48 h后,分别采用CCK-8、EdU及划痕实验检测细胞活力、增殖及迁移能力。结果 Gal-3在ICC组织中显著高表达,且与预后不良相关。虚拟筛选确定Magnesium Lithospermate B（MLB）为潜在Gal-3小分子抑制剂,体外实验验证其可显著抑制ICC细胞的增殖和迁移。结论 Gal-3在ICC中高表达,MLB可作为其有效抑制剂,对ICC细胞具有抑制作用,具有潜在的抗肿瘤价值。
Abstract
Objective This study aimed to identify small-molecule inhibitors targeting Galectin-3(Gal-3) based on machine learning techniques and to investigate their binding modes with Gal-3 as well as their effects on the proliferation and migration of intrahepatic cholangiocarcinoma (ICC) cells. Methods Bioinformatics and immunohistochemistry were used to analyze Gal-3 expression in ICC tissues and its correlation with prognosis. The structure of Gal-3 and the TargetMol L4000 compound library were preprocessed using Schrödinger software, followed by virtual screening through molecular docking. Candidate inhibitors were selected via clustering analysis and manual evaluation. ICC cell lines were treated with different concentrations of the inhibitors for 24 h and 48 h, and CCK-8, EdU, and wound healing assays were performed to assess cell viability, proliferation, and migration. Results Gal-3 was significantly overexpressed in ICC tissues and correlated with poor prognosis. Magnesium Lithospermate B (MLB) was identified as a potential Gal-3 inhibitor through virtual screening. In vitro experiments showed that MLB significantly inhibited ICC cell proliferation and migration. Conclusion Gal-3 is highly expressed in ICC and may serve as a therapeutic target. MLB, as a Gal-3 inhibitor, exhibits potential antitumor activity against ICC.
关键词
肝内胆管癌 /
半乳糖凝集素-3 /
小分子抑制剂 /
MLB
Key words
intrahepatic cholangiocarcinoma /
Gal-3 /
molecule inhibitors /
MLB
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参考文献
[1] ELIAS C, RAHMAN A, MIAL-ANTHONY J, et al.Advancements in cholangiocarcinoma: evolving strategies for diagnosis, treatment, and palliation over three decades[J]. Chin Clin Oncol, 2024, 13(5): 70.
[2] SARCOGNATO S, SACCHI D, FASSAN M, et al.Cholangiocarcinoma[J]. Pathologica, 2021, 113(3): 158-169.
[3] MOCAN L P, RUSU I, MELINCOVICI C S, et al.The Role of Immunohistochemistry in the Differential Diagnosis between Intrahepatic Cholangiocarcinoma, Hepatocellular Carcinoma and Liver Metastasis, as Well as Its Prognostic Value[J]. Diagnostics (Basel), 2023, 13(9): 1542.
[4] ZHANG H, YANG T, WU M, et al.Intrahepatic cholangiocarcinoma: Epidemiology, risk factors, diagnosis and surgical management[J]. Cancer Lett, 2016, 379(2): 198-205.
[5] ALQAHTANI S A, PAIK J M, BISWAS R, et al.Poor Awareness of LiverDisease Among Adults With NAFLD in the United States[J]. Hepatol Commun, 2021, 5(11): 1833-1847.
[6] JEETHY RAM T, LEKSHMI A, SOMANATHAN T, et al.Galectin-3: A factotum in carcinogenesis bestowing an archery for prevention[J]. Tumour Biol, 2021, 43(1): 77-96.
[7] DIMITRIJEVIC STOJANOVIC M, STOJANOVIC B, RADOSAVLJEVIC I, et al.Galectin-3’s Complex Interactions in Pancreatic Ductal Adenocarcinoma: From Cellular Signaling to Therapeutic Potential[J]. Biomolecules, 2023, 13(10): 1500.
[8] GUAN C, LIU L, ZHAO Y, et al.YY1 and eIF4A3 are mediators of the cell proliferation, migration and invasion in cholangiocarcinoma promoted by circ-ZNF609 by targeting miR-432-5p to regulate LRRC1[J]. Aging (Albany NY), 2021, 13(23): 25195-25212.
[9] QIU B, CHEN T, SUN R, et al.Sprouty4 correlates with favorable prognosis in perihilar cholangiocarcinoma by blocking the FGFR-ERK signaling pathway and arresting the cell cycle[J]. EBioMedicine, 2019, 50: 166-177.
[10] AHN C S, KIM J G, KANG I, et al.Omega-Class Glutathione Transferases of Carcinogenic Liver Fluke, Clonorchis sinensis, Modulate Apoptosis and Differentiation of Host Cholangiocytes[J]. Antioxidants (Basel), 2021, 10(7): 1017.
[11] CHEN J, PAN Q, BAI Y, et al.Hydroxychloroquine Induces Apoptosis in Cholangiocarcinoma via Reactive Oxygen Species Accumulation Induced by Autophagy Inhibition[J]. Front Mol Biosci, 2021, 8: 720370.
[12] CROSBY D.Delivering on the promise of early detection with liquid biopsies[J]. Br J Cancer, 2022, 126(3): 313-315.
[13] LU H Y, SHIH C M, HUANG C Y, et al.Galectin-3 Modulates Macrophage Activation and Contributes Smooth Muscle Cells Apoptosis in Abdominal Aortic Aneurysm Pathogenesis[J]. Int J Mol Sci, 2020, 21(21): 8257.
[14] LI T, ZHA L, LUO H, et al.Galectin-3 Mediates Endothelial-to-Mesenchymal Transition in Pulmonary Arterial Hypertension[J]. Aging Dis, 2019, 10(4): 731-745.
[15] FAN C, QIN K, IROEGBU C D, et al.Magnesium lithospermate B enhances the potential of human-induced pluripotent stem cell-derived cardiomyocytes for myocardial repair[J]. Chin Med J (Engl), 2024, 137(15): 1857-1869.
[16] ZHANG N, HAN L, XUE Y, et al.The Protective Effect of Magnesium Lithospermate B on Hepatic Ischemia/Reperfusion via Inhibiting the Jak2/Stat3 Signaling Pathway[J]. Front Pharmacol, 2019, 10: 620.
[161 ZHAO Q, CHENG X, GUO J, et al. MLKL inhibits intestinal tumorigenesis by suppressing STAT3 signaling pathway[J]. Int J Biol Sci, 2021, 17(3): 869-881.
[18] REZENDE C P, MARTINS OLIVEIRA BRITO P K, PESSONI A M, et al. Altered expression of genes related to innate antifungal immunity in the absence of galectin-3[J]. Virulence, 2021, 12(1): 981-988.
[19] ZUO X L, CHEN Z Q, WANG J F, et al.miR-337-3p suppresses the proliferation and invasion of hepatocellular carcinoma cells through targeting JAK2[J]. Am J Cancer Res, 2018, 8(4): 662-674.
[20] ECKARDT V, MILLER M C, BLANCHET X, et al.Chemokines and galectins form heterodimers to modulate inflammation[J]. EMBO Rep, 2020, 21(4): e47852.
基金
湖南省卫生健康高层次人才重大科研专项“Galectin-8调控cGAS-STING信号通路在ICC中的作用及机制研究”（20230541）; 区域遗传性出生缺陷防控研究湖南省重点实验室开放课题重点项目“SFXN1介导半乳糖凝集素3促进妊娠期糖尿病进展的机制研究”（HPKL2023003）
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