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Sun Young Park,1 Beomjin Kim,2 Zhengwei Cui,3 Geuntae Park,4 Young-Whan Choi3 1Bio-IT Fusion Technology Research Institute, Pusan National University, Busan 609-735, Korea; 2Department of Nanomaterials Engineering, Pusan National University, Busan 609-735, Korea; 3Department of Horticultural Bioscience, Pusan National University, Myrang 627-706, Korea; 4Department of Nanofusion Technology, Graduate School, Pusan National University, Busan, 609-735, KoreaCorrespondence: Geuntae ParkDepartment of Nanofusion Technology, Graduate School, Pusan National University, Busan 609-735, Republic of KoreaTel +82-51-510-3740Fax +82-51-518-4113Email gtpark@pusan.ac.krYoung-Whan ChoiDepartment of Horticultural Bioscience, Pusan National University, Miryang 627-706, Republic of KoreaTel +82-55-350-5522Fax +82-55-350-5529Email ywchoi@pusan.ac.krPurpose: We aimed to study green-synthesized gold nanoparticles (GNPs) from Maclura tricuspidata (MT) root (MTR), stem (MTS), leaf (MTL), and fruit (MTF) extracts and evaluate their anti-metastatic properties in hepatocellular carcinoma cells. Maclura tricuspidata belongs to the Moraceae family and is widely used as a traditional medicinal plant given its biological activities.Methods: We quantified the phenolic and flavonoid contents, reducing capacity, and antioxidant activity of all four extracts. The facile and optimum synthesis of MT-GNPs was visualized using UV-vis spectra and dynamic light scattering (DLS). Surface morphology, selected area electron diffraction (SAED), and fast Fourier transform (FFT) pattern of MT-GNPs were assessed using high-resolution transmission electron microscopy (HR-TEM). The crystallized gold pattern of MT-GNPs was evaluated using energy dispersive spectroscopy (EDS) and X-ray diffraction (XRD). The functionalizing ligands of MT-extracts and MT-GNPs were determined using Fourier-transform infrared spectroscopy (FT-IR). The photocatalytic capabilities of MT-GNPs were assessed by measuring the reduction of rhodamine B and methylene blue. Cell viability assay was detected using Cell Counting Kit-8 solution. Anti-migratory and anti-invasive effects were assessed using cell migration and invasion assays. Matrix metalloproteinase (MMP)-9 and phospholipase D (PLD) enzymatic activities were measured using gelatin zymography and Amplex Red PLD assay, respectively. Western blotting and luciferase assay were used to detect protein expression.Results: All extracts had high phenolic and flavonoid contents and strong antioxidant and reducing capacities. Results from UV-Vis spectra, DLS, HR-TEM, EDS, XRD, and FT-IR showed the successful formation of MT-GNP with surface morphology, crystallinity, reduction capacity, capsulation, and stabilization. MTR-GNPs and MTS-GNPs had better catalytic activities than MTL-GNPs and MTF-GNPs for reduction of methylene blue and rhodamine B. Moreover, MTS-GNPs and MTR-GNPs exhibited the highest anti-migratory and anti-invasive potential and seemed to be more biologically active than the MTS and MTR extracts. Treatment with MT-GNPs decreased the enzymatic activity, translation levels of MMP-9 and PLD1. Our results showed that MTS-GNPs and MTR-GNPs could dramatically reverse transforming growth factor-β-induced vimentin and N-cadherin upregulation and E-cadherin downregulation.Conclusion: The application of GNPs as a potential treatment approach for hepatocellular carcinoma can improve therapeutic efficiency.Keywords: MT-GNPs, Maclura tricuspidata, hepatocellular carcinoma cells, epithelial–mesenchymal transition