title: Co-delivery of VP-16 and Bcl-2-targeted antisense on PEG-grafted oMWCNTs for synergistic in vitro anti-cancer effects in non-small and small cell lung cancer.
abstract: Present study describes the preparation of a polyethylene glycol-grafted oxidized multi-walled carbon nanotubes (oMWCNTs-PEG) hybrid nanosystem as a carrier of etoposide (VP-16) and Bcl-2 phosphorothioate antisense deoxyoligonucleotides (Aso) to achieve a superior cytostastic efficacy in non-small and small cell lung cancer in vitro. We have demonstrated that the adsorption of hydrophobic VP-16 and Bcl-2 Aso results in a stable nanotransporter exhibiting good dispersion with excellent release profiles (both, in pH 7.4 and 4.8) and negligible hemolytic activity (up to 6.5%). The evaluation of cytotoxicity was carried out in in vitro using small cell (SCLC; DMS53) and non-small cell lung cancer (NSCLC; NCIH2135) cell lines. It was found that Bcl-2 interference significantly increased the anti-cancer efficiency of VP-16 in the chemoresistant NSCLC cells. This was further supported using a flow-cytometry (Annexin V/propidium iodide assay), which revealed a significant increase in apoptotic cells in both the cell lines after the co-administration of VP-16 and Bcl-2 Aso using oMWCNTs-PEG hybrid, and fluorescence microscopy, which showed an increase in reactive oxygen species identified after Bcl-2 knock-down. Overall, oMWCNTs-PEG provided an exceptional biocompatible vehicle enabling the internalization of negatively charged nucleic acids and pH-sensitive release of cargoes in a hypoxic environment of the most of solid tumors. Moreover, Aso specifically binding to the first six codons of the Bcl-2 mRNA gave a satisfactorily decrease in Bcl-2 translation and an increase in NCIH2135 chemosensitivity towards VP-16.
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czechia; Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia.
authors.forename: Zbynek
authors.initials: Z
authors.lastname: Heger
authors.affiliation: Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia; Department of Physiology and Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Kamenice 5, CZ-625 00 Brno, Czechia.
authors.forename: Hana
authors.initials: H
authors.lastname: Polanska
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czechia; Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia.
authors.forename: Sona
authors.initials: S
authors.lastname: Krizkova
authors.affiliation: Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia; Department of Physiology and Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Kamenice 5, CZ-625 00 Brno, Czechia.
authors.forename: Jan
authors.initials: J
authors.lastname: Balvan
authors.affiliation: Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia; Department of Physiology and Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Kamenice 5, CZ-625 00 Brno, Czechia.
authors.forename: Martina
authors.initials: M
authors.lastname: Raudenska
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czechia; Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia.
authors.forename: Simona
authors.initials: S
authors.lastname: Dostalova
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czechia; Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia.
authors.forename: Amitava
authors.initials: A
authors.lastname: Moulick
authors.affiliation: Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia; Department of Physiology and Department of Pathological Physiology, Faculty of Medicine, Masaryk University, Kamenice 5, CZ-625 00 Brno, Czechia.
authors.forename: Michal
authors.initials: M
authors.lastname: Masarik
authors.affiliation: Department of Chemistry and Biochemistry, Mendel University in Brno, Zemedelska 1, CZ-613 00 Brno, Czechia; Central European Institute of Technology, Brno University of Technology, Purkynova 123, CZ-612 00 Brno, Czechia. Electronic address: vojtech.adam@mendelu.cz.
authors.forename: Vojtech
authors.initials: V
authors.lastname: Adam
mesh_terms: D000970:Antineoplastic Agents / Q000008:administration & dosage*; D017209:Apoptosis; D001672:Biocompatible Materials / Q000737:chemistry; D002289:Carcinoma, Non-Small-Cell Lung / Q000378:metabolism* / Q000473:pathology; D015687:Cell Hypoxia; D045744:Cell Line, Tumor / Q000187:drug effects; D004337:Drug Carriers; D004357:Drug Synergism; D004586:Electrophoresis; D004912:Erythrocytes / Q000166:cytology; D005047:Etoposide / Q000008:administration & dosage*; D005434:Flow Cytometry; D006461:Hemolysis; D006801:Humans; D006863:Hydrogen-Ion Concentration; D057927:Hydrophobic and Hydrophilic Interactions; D007700:Kinetics; D008175:Lung Neoplasms / Q000378:metabolism* / Q000473:pathology; D008855:Microscopy, Electron, Scanning; D037742:Nanotubes, Carbon / Q000737:chemistry; D009696:Nucleic Acids / Q000737:chemistry; D009841:Oligonucleotides / Q000737:chemistry; D016376:Oligonucleotides, Antisense / Q000737:chemistry; D011092:Polyethylene Glycols / Q000737:chemistry; D019253:Proto-Oncogene Proteins c-bcl-2 / Q000235:genetics*; D017382:Reactive Oxygen Species / Q000737:chemistry; D055752:Small Cell Lung Carcinoma / Q000378:metabolism* / Q000473:pathology
chemical_list: D000970:Antineoplastic Agents; D001672:Biocompatible Materials; D004337:Drug Carriers; D037742:Nanotubes, Carbon; D009696:Nucleic Acids; D009841:Oligonucleotides; D016376:Oligonucleotides, Antisense; D019253:Proto-Oncogene Proteins c-bcl-2; D017382:Reactive Oxygen Species; D011092:Polyethylene Glycols; D005047:Etoposide
keywords: Drug delivery; Etoposide; In vitro release; Multi-walled carbon nanotubes; Nanomedicine