title: iTRAQ-based proteomic identification of proteins involved in anti-angiogenic effects of Panduratin A on HUVECs.
abstract: Panduratin A (PA), a cyclohexanyl chalcone from Boesenbergia rotunda (L.) Mansf. was shown to possess anti-angiogenic effects in our previous study. In the present study, the molecular targets and anti-angiogenic mechanisms of PA on human umbilical vein endothelial cells (HUVECs) were identified using an iTRAQ-based quantitative proteomics approach. A total of 263 proteins were found to be differentially regulated in response to treatment with PA. Ingenuity Pathway Analysis revealed that cellular growth and proliferation, protein synthesis, RNA post-transcriptional modification, cellular assembly and organization and cell-to-cell signaling and interaction were the most significantly deregulated molecular and cellular functions in PA-treated HUVECs. PA inhibited the expressions of ARPC2 and CTNND1 that are associated with the formation of actin cytoskeleton, focal adhesion and cellular protrusions. In addition, PA down-regulated CD63, GRB-2, ICAM-2 and STAB-1 that are implicated in adhesion, migration and tube formation of endothelial cells. The differential expressions of three targets, namely, ARPC2, CDK4, and GRB-2 were validated by western blot analyses. Furthermore, PA inhibited G1-S progression, and resulted in G0/G1 arrest in HUVECs. The blockage in cell cycle progression was accompanied with the suppression of mTOR signaling. Treatment of HUVECs with PA resulted in decreased phosphorylation of ribosomal S6 and 4EBP1 proteins, the two downstream effectors of mTOR signaling. We further showed that PA is able to inhibit mTOR signaling induced by VEGF, a potent inducer of angiogenesis. Taken together, by integrating quantitative proteomic approach, we identified protein targets in which PA mediates its anti-angiogenic effects. The present study thus provides mechanistic evidence to the previously reported multifaceted anti-angiogenic effects of PA. Our study further identified mTOR signaling as an important target of PA, and therefore highlights the potential of PA for therapeutic intervention against angiogenesis-related pathogenesis, particularly, metastatic malignancy.
authors: Centre of Natural Products & Drug Discovery (CENAR), Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors: Siew-Li
authors: SL
authors: Lai
authors: Centre of Natural Products & Drug Discovery (CENAR), Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia. Electronic address: wongpf@um.edu.my.
authors: Pooi-Fong
authors: PF
authors: Wong
authors: Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore.
authors: Teck-Kwang
authors: TK
authors: Lim
authors: Department of Biological Sciences, National University of Singapore, 14 Science Drive 4, Singapore 117543, Singapore.
authors: Qingsong
authors: Q
authors: Lin
authors: Centre of Natural Products & Drug Discovery (CENAR), Department of Pharmacology, Faculty of Medicine, University of Malaya, 50603 Kuala Lumpur, Malaysia.
authors: Mohd Rais
authors: MR
authors: Mustafa
mesh_terms: D051376:Actin-Related Protein 2-3 Complex / Q000378:metabolism; D020533:Angiogenesis Inhibitors / Q000494:pharmacology*; D049109:Cell Proliferation / Q000187:drug effects; D047188:Chalcones / Q000494:pharmacology*; D051358:Cyclin-Dependent Kinase 4 / Q000378:metabolism; D051380:GRB2 Adaptor Protein / Q000378:metabolism; D061307:Human Umbilical Vein Endothelial Cells / Q000187:drug effects*; D006801:Humans; D020543:Proteome / Q000378:metabolism*; D040901:Proteomics; D027343:Rhizome / Q000737:chemistry; D030521:Zingiberaceae / Q000737:chemistry
chemical_list: C000595669:ARPC2 protein, human; D051376:Actin-Related Protein 2-3 Complex; D020533:Angiogenesis Inhibitors; D047188:Chalcones; D051380:GRB2 Adaptor Protein; C495948:GRB2 protein, human; D020543:Proteome; C482884:panduratin A; C495900:CDK4 protein, human; D051358:Cyclin-Dependent Kinase 4
keywords: Anti-angiogenesis; Panduratin A; Proteomics; iTRAQ