title: Therapeutic implication of 'Iturin A' for targeting MD-2/TLR4 complex to overcome angiogenesis and invasion.
abstract: Tumor angiogenesis and invasion are deregulated biological processes that drive multistage transformation of tumors from a benign to a life-threatening malignant state activating multiple signaling pathways including MD-2/TLR4/NF-κB. Development of potential inhibitors of this signaling is emerging area for discovery of novel cancer therapeutics. In the current investigation, we identified Iturin A (A lipopeptide molecule from Bacillus megaterium) as a potent inhibitor of angiogenesis and cancer invasion by various in vitro and in vivo methods. Iturin A was found to suppress VEGF, a powerful inducer of angiogenesis and key player in tumor invasion, as confirmed by ELISA, western blot and real time PCR. Iturin A inhibited endothelial tube arrangement, blood capillary formation, endothelial sprouting and vascular growth inside the matrigel. In addition, Iturin A inhibited MMP-2/9 expression in MDA-MB-231 and HUVEC cells. Cancer invasion, migration and colony forming ability were significantly hampered by Iturin A. Expressions of MD-2/TLR4 and its downstream MyD88, IKK-α and NF-κB were also reduced in treated MDA-MB-231 and HUVEC cells. Western blot and immunofluorescence study showed that nuclear accumulation of NF-κB was hampered by Iturin A. MD-2 siRNA or plasmid further confirmed the efficacy of Iturin A by suppressing MD-2/TLR4 signaling pathway. The in silico docking study showed that the Iturin A interacted well with the MD-2 in MD-2/TLR4 receptor complex. Conclusively, inhibition of MD-2/TLR4 complex with Iturin A offered strategic advancement in cancer therapy.
authors.affiliation: School of Medical Science & Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
authors.forename: Goutam
authors.initials: G
authors.lastname: Dey
authors.affiliation: School of Medical Science & Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
authors.forename: Rashmi
authors.initials: R
authors.lastname: Bharti
authors.affiliation: Drug Theoretics and Cheminformatics Laboratory, Division of Medicinal and Pharmaceutical Chemistry, Department of Pharmaceutical Technology, Jadavpur University, Kolkata 700032, India.
authors.forename: Probir Kumar
authors.initials: PK
authors.lastname: Ojha
authors.affiliation: School of Medical Science & Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
authors.forename: Ipsita
authors.initials: I
authors.lastname: Pal
authors.affiliation: School of Medical Science & Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
authors.forename: Y
authors.initials: Y
authors.lastname: Rajesh
authors.affiliation: Division of Nuclear Medicine, Indian Institute of Chemical Biology, Kolkata 700032, West Bengal, India.
authors.forename: Indranil
authors.initials: I
authors.lastname: Banerjee
authors.affiliation: School of Medical Science & Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
authors.forename: Payel
authors.initials: P
authors.lastname: Banik
authors.affiliation: School of Medical Science & Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
authors.forename: Sheetal
authors.initials: S
authors.lastname: Parida
authors.affiliation: School of Medical Science & Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India.
authors.forename: Aditya
authors.initials: A
authors.lastname: Parekh
authors.affiliation: Department of Biotechnology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India. Electronic address: rksen@yahoo.com.
authors.forename: Ramkrishna
authors.initials: R
authors.lastname: Sen
authors.affiliation: School of Medical Science & Technology, Indian Institute of Technology Kharagpur, Kharagpur 721302, India. Electronic address: mahitosh@smst.iitkgp.ernet.in.
authors.forename: Mahitosh
authors.initials: M
authors.lastname: Mandal
mesh_terms: D001410:Bacillus megaterium / Q000737:chemistry; D045744:Cell Line, Tumor; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D061307:Human Umbilical Vein Endothelial Cells; D006801:Humans; D051550:I-kappa B Kinase / Q000235:genetics; D051218:Lymphocyte Antigen 96 / Q000737:chemistry / Q000235:genetics*; D053594:Myeloid Differentiation Factor 88 / Q000235:genetics; D016328:NF-kappa B / Q000235:genetics; D009361:Neoplasm Invasiveness / Q000235:genetics; D009369:Neoplasms / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000235:genetics / Q000473:pathology; D010456:Peptides, Cyclic / Q000008:administration & dosage* / Q000737:chemistry; D034741:RNA, Small Interfering / Q000235:genetics; D015398:Signal Transduction / Q000187:drug effects; D051197:Toll-Like Receptor 4 / Q000737:chemistry / Q000235:genetics*
chemical_list: C495377:LY96 protein, human; D051218:Lymphocyte Antigen 96; C507779:MYD88 protein, human; D053594:Myeloid Differentiation Factor 88; D016328:NF-kappa B; D010456:Peptides, Cyclic; D034741:RNA, Small Interfering; C495345:TLR4 protein, human; D051197:Toll-Like Receptor 4; C013579:iturin A; C496559:CHUK protein, human; D051550:I-kappa B Kinase
keywords: Invasion; MD-2 (myeloid differentiation factor 2); TLR4; Tumor angiogenesis; Vascular endothelial growth factor (VEGF) and human umbilical vein endothelial cell (HUVEC)