title: Identification of possible cytotoxicity mechanism of polyethylenimine by proteomics analysis.
abstract: Polyethylenimine (PEI) is a polycation widely used for successful gene delivery both in vitro and in vivo experiments. However, different studies showed that PEI could be cytotoxic to transfected cells, and the mechanism of toxicity is poorly understood. Identification of PEI-interacting proteins may help in understanding the toxicity pathways. In this study, we investigated proteins that could interact with PEI in human colorectal adenocarcinoma cells (HT29). In order to identify the proteins interacting with PEI, PEI was immobilized to sepharose beads as solid matrix. The HT29 cell lysate were passed through the matrix. PEI-bound proteins were isolated, and further separation was performed by two-dimensional gel electrophoresis. After gel digestion, proteins were identified by matrix-assisted laser desorption/ionization-time-of-flight (TOF)/TOF mass spectrometry. Our data indicated that most of the identified PEI-interacting proteins such as shock proteins, glutathione-S-transferases, and protein disulfide isomerase are involved in apoptosis process in cells. Thus, although this is a preliminary experiment implicating the involvement of some proteins in PEI cytotoxicity, it could partly explain the mechanism of PEI cytotoxicity in cells.
authors.affiliation: Nanotechnology Research Center, Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: M
authors.initials: M
authors.lastname: Khansarizadeh
authors.affiliation: School of Medicine, Gonabad University of Medical Sciences, Gonabad, Iran Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: A
authors.initials: A
authors.lastname: Mokhtarzadeh
authors.affiliation: Department of Pharmacology and Toxicology, School of Pharmacy, Shiraz University of Medical Sciences, Shiraz, Iran.
authors.forename: M
authors.initials: M
authors.lastname: Rashedinia
authors.affiliation: Targeted Drug Delivery Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: S M
authors.initials: SM
authors.lastname: Taghdisi
authors.affiliation: Department of Pharmacodynamy and Toxicology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: P
authors.initials: P
authors.lastname: Lari
authors.affiliation: Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran.
authors.forename: K H
authors.initials: KH
authors.lastname: Abnous
authors.affiliation: Nanotechnology Research Center, Department of Pharmaceutical Biotechnology, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran Pharmaceutical Research Center, School of Pharmacy, Mashhad University of Medical Sciences, Mashhad, Iran ramezanim@mums.ac.ir.
authors.forename: M
authors.initials: M
authors.lastname: Ramezani
mesh_terms: D002470:Cell Survival / Q000187:drug effects*; D005786:Gene Expression Regulation / Q000187:drug effects; D019073:HT29 Cells; D006801:Humans; D011094:Polyethyleneimine / Q000633:toxicity*; D040901:Proteomics / Q000379:methods*
chemical_list: D011094:Polyethyleneimine
keywords: 2-D gel electrophoresis; MALDI-TOF/TOF; Polyethylenimine (PEI); gene delivery; toxicity