title: T11TS Treatment Augments Apoptosis of Glioma Associated Brain Endothelial Cells, Hint Toward Anti-Angiogenic Action in Glioma.
abstract: Malignant glioma continues to be a clinical challenge with an urgent need for developing curative therapeutic intervention. Apoptosis induction in tumor-associated endothelial cells represent a central mechanism that counteracts angiogenesis in glioma and other solid tumors. We previously demonstrated that intraperitoneal administration of sheep erythrocyte membrane glycopeptide T11-target structure (T11TS) in rodent glioma model inhibits PI3K/Akt pathway and Raf/MEK/ERK signaling in glioma-associated brain endothelial cells. In the present study, we investigated whether T11TS treatment influence apoptosis signaling in vivo in glioma-associated brain endothelial cells. Annexin-V/PI staining showed that T11TS treatment in glioma-induced rats increases apoptosis of glioma-associated endothelial cells within glioma milieu compared to brain endothelial cells in glioma induced and control groups. Flowcytometric JC-1 assay revealed that T11TS administration triggers loss of mitochondrial membrane potential in glioma-associated brain endothelial cells. Flowcytometry, immunoblotting, and in situ immunofluoresecnt imaging were employed to investigate the effect of T11TS on apoptotic regulatory proteins in brain endothelial cells. T11TS treatment-upmodulated expression of p53, Bax, Fas, FasL, and FADD in glioma associated endothelial cells and downregulated Bcl-2 protein. T11TS therapy induced cytochrome-c release into cytosol, activated caspase -9, 8, 3, and cleaved Bid in glioma associated brain endothelial cells. The study demonstrates that T11TS induces apoptosis in glioma-associated brain endothelial cells via p53 accumulation and activation of intrinsic as well as Fas-dependent extrinsic pathway. The pro-apoptotic action of T11TS on glioma-associated endothelial cells provides crucial insight into how T11TS exerts its anti-angiogenic function in glioma. J. Cell. Physiol. 232: 526-539, 2017. © 2016 Wiley Periodicals, Inc.
authors.affiliation: Cellular and Molecular Immunology Laboratory, Department of Laboratory Medicine, School of Tropical Medicine, Kolkata, India.|Laboratory of Molecular Neuro-Oncology, Department of Neurosurgery, Winship Cancer Institute of Emory University, Atlanta, Georgia.
authors.forename: Debanjan
authors.initials: D
authors.lastname: Bhattacharya
authors.affiliation: Cellular and Molecular Immunology Laboratory, Department of Laboratory Medicine, School of Tropical Medicine, Kolkata, India.
authors.forename: Manoj Kumar
authors.initials: MK
authors.lastname: Singh
authors.affiliation: Cellular and Molecular Immunology Laboratory, Department of Laboratory Medicine, School of Tropical Medicine, Kolkata, India.
authors.forename: Suhnrita
authors.initials: S
authors.lastname: Chaudhuri
authors.affiliation: Cellular and Molecular Immunology Laboratory, Department of Laboratory Medicine, School of Tropical Medicine, Kolkata, India.
authors.forename: Ankur
authors.initials: A
authors.lastname: Datta
authors.affiliation: Cellular and Molecular Immunology Laboratory, Department of Laboratory Medicine, School of Tropical Medicine, Kolkata, India.
authors.forename: Swapna
authors.initials: S
authors.lastname: Chaudhuri
mesh_terms: D020533:Angiogenesis Inhibitors / Q000494:pharmacology / Q000627:therapeutic use*; D000818:Animals; D017209:Apoptosis / Q000187:drug effects*; D051023:BH3 Interacting Domain Death Agonist Protein / Q000378:metabolism; D001932:Brain Neoplasms / Q000188:drug therapy* / Q000473:pathology; D020169:Caspases / Q000378:metabolism; D045304:Cytochromes c / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D042783:Endothelial Cells / Q000187:drug effects / Q000473:pathology*; D004789:Enzyme Activation; D053222:Fas Ligand Protein / Q000378:metabolism; D053200:Fas-Associated Death Domain Protein / Q000378:metabolism; D005260:Female; D005910:Glioma / Q000473:pathology*; D006020:Glycopeptides / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use*; D008297:Male; D008562:Membrane Glycoproteins / Q000008:administration & dosage / Q000494:pharmacology* / Q000627:therapeutic use*; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D008954:Models, Biological; D009389:Neovascularization, Pathologic / Q000188:drug therapy* / Q000473:pathology; D051381:Rats; D012756:Sheep; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism; D051028:bcl-2-Associated X Protein / Q000378:metabolism; D019014:fas Receptor / Q000378:metabolism
chemical_list: D020533:Angiogenesis Inhibitors; D051023:BH3 Interacting Domain Death Agonist Protein; C505535:Fadd protein, rat; D053222:Fas Ligand Protein; C491898:Fas protein, rat; D053200:Fas-Associated Death Domain Protein; D006020:Glycopeptides; D008562:Membrane Glycoproteins; D016159:Tumor Suppressor Protein p53; D051028:bcl-2-Associated X Protein; D019014:fas Receptor; D045304:Cytochromes c; D020169:Caspases