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Hypoxia-inducible bidirectional shRNA expression vector delivery using PEI/chitosan-TBA copolymers for colorectal Cancer gene therapy

Javan, B. and Atyabi, F. and Shahbazi, M. (2018) Hypoxia-inducible bidirectional shRNA expression vector delivery using PEI/chitosan-TBA copolymers for colorectal Cancer gene therapy. Life Sciences, 202. pp. 140-151.

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Abstract

Aims: This investigation was conducted to construct a hypoxia/colorectal dual-specific bidirectional short hairpin RNA (shRNA) expression vector and to transfect it into the colon cancer cell line HT-29 with PEI/chitosan-TBA nanoparticles for the simultaneous knock down of β-catenin and Bcl-2 under hypoxia. Main methods: To construct a pRNA-bipHRE-CEA vector, the carcinoma embryonic antigen (CEA) promoter designed in two directions and the vascular endothelial growth factor (VEGF) enhancer were inserted between two promoters for hypoxic cancer specific gene expression. To confirm the therapeutic effect of the dual-specific vector, β-catenin and Bcl-2 shRNAs were inserted downstream of each promoter. The physicochemical properties, the cytotoxicity, and the transfection efficiency of these PEI/chitosan-TBA nanoparticles were investigated. In addition, the antitumor effects of the designed vector on the expression of β-catenin and Bcl-2, cell cycle distribution, and apoptosis were investigated in vitro. Key findings: The silencing effect of the hypoxia-response shRNA expression vector was relatively low (18–25) under normoxia, whereas it was significantly increased to approximately 50–60 in the HT-29 cell line. Moreover, the cancer cells showed significant G0/G1 arrest and increased apoptosis due to gene silencing under hypoxia. Furthermore, MTS assay, fluorescence microscopy images, and flow cytometry analyses confirmed that the PEI/chitosan-TBA blend system provided effective transfection with low cytotoxicity. Significance: This novel hypoxia-responsive shRNA expression vector may be useful for RNA interference (RNAi)-based cancer gene therapy in hypoxic colorectal tumors. Moreover, the PEI/chitosan-TBA copolymer might be a promising gene carrier for use in gene transfer in vivo. © 2018

Item Type: Article
Additional Information: cited By 0
Uncontrolled Keywords: beta catenin; carcinoembryonic antigen; chitosan; copolymer; nanoparticle; polyethyleneimine; protein bcl 2; short hairpin RNA; vasculotropin; beta catenin; chitosan; chitosan 2-iminothiolane; polymer; small interfering RNA; thiol derivative, antineoplastic activity; apoptosis; Article; cancer gene therapy; cell cycle; cell hypoxia; colorectal cancer; controlled study; drug cytotoxicity; flow cytometry; fluorescence microscopy; G1 phase cell cycle checkpoint; gene delivery system; gene expression; gene vector; HT-29 cell line; human; human cell; in vitro study; physical chemistry; promoter region; RNA transport; administration and dosage; analogs and derivatives; biosynthesis; colorectal tumor; gene therapy; gene transfer; genetics; hypoxia; metabolism; particle size; procedures; proto oncogene, Apoptosis; beta Catenin; Cell Cycle; Chitosan; Colorectal Neoplasms; Gene Transfer Techniques; Genes, bcl-2; Genetic Therapy; Genetic Vectors; HT29 Cells; Humans; Hypoxia; Nanoparticles; Particle Size; Polymers; RNA, Small Interfering; Sulfhydryl Compounds
Subjects: مقالات نمایه شده محققین دانشگاه در سایت ,Web of Science ,Scopus
Divisions: UNSPECIFIED
Depositing User: GOUMS
Date Deposited: 26 Jun 2018 08:12
Last Modified: 26 Jun 2018 08:12
URI: http://eprints.goums.ac.ir/id/eprint/9596

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