Neferine induced autophagy and inhibited migration through FAK and S6K1 signaling pathways in brain tumor

碩士 === 國立東華大學 === 生命科學系 === 103 === Focal adhesion kinase (FAK) and human p70 ribosomal S6 kinase (S6K1), non-receptor protein tyrosine, have been overexpressed in many types of tumors and play an important role of cell signaling pathways including cell migration, proliferation, viability, and cell...

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Bibliographic Details
Main Authors: Yu-Chuan Chang, 張郁涓
Other Authors: Ching-Feng Weng
Format: Others
Published: 2015
Online Access:http://ndltd.ncl.edu.tw/handle/13659782704960498417
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Summary:碩士 === 國立東華大學 === 生命科學系 === 103 === Focal adhesion kinase (FAK) and human p70 ribosomal S6 kinase (S6K1), non-receptor protein tyrosine, have been overexpressed in many types of tumors and play an important role of cell signaling pathways including cell migration, proliferation, viability, and cell survival. This study was aimed to screen the novel and specific inhibitors of FAK and S6K1 from natural compounds via molecular docking and further investigate the underlying mechanism of action. The 3D structures of FAK (PDB ID: 2AL6) and S6K1 (PDB ID: 3A60) were employed to do docking with 60 natural compounds. Based on their high affinity and energy interaction, top one docking score compound Neferine from Nelumbo Nucifera Gaertn was selected and further validated the anti-tumor effect in C6 rat Glioma and IMR32 human Neuroblastoma cells. FAK (2AL6)-ligands interaction analysis indicated that H-bond with residues Arg86, Arg125, and Tyr251 while S6K1 (3A60)-ligands interaction analysis indicated that H-bond with residues Arg95, Gly100, and Asp236. Neferine showed a potential effect on cell viability by MTT assay and arrest the cell cycle at G2/M phase by flow cytometer. The protein levels of FAK, pFAK, S6K1, and pS6K1 were decreased, which are the key regulators cell migration and proliferation, respectively. Neferine induced the autophagy through Beclin-1 and LC3 pathway, and induced the apoptosis through caspase 3 and PARP. Furthermore, Neferine would inhibit the cell migration on C6 cells by wound healing assay. Taken altogether, this study suggests that Neferine could be a potential inhibitor of FAK and S6K1 for anti-tumorigenesis and anti-metastasis.