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N-Isopropylacrylamide-modified polyethylenimine-mediated p53 gene delivery to prevent the proliferation of cancer cells.

Abstract
In this paper, N-isopropylacrylamide-modified polyethylenimine (PEN) was constructed through Michael addition and employed as a carrier to achieve the p53 gene delivery, using HeLa (p53wt) and PC-3 cells (p53null) as models. After PEN-mediated p53 transfection, expression level of p53 in HeLa and PC3 cells was up-regulated at both mRNA and protein levels. Due to the exogenous p53 expression, the inhibition of cell proliferation was observed through MTT analysis, attributing to the activation of apoptosis and cell cycle arrest. Using flow cytometric analysis, early apoptotic ratios of 54.95% and 27.06% after PEN-mediated p53 transfection were detected in PC-3 and HeLa cells, respectively, indicating that PC-3 cells were more sensitive to the exogenous p53 transfection than HeLa cells. Meanwhile, G1 phase arrest was detected in PC-3 cells while a unique G2 phase arrest was identified in HeLa cells after p53 transfection. Through Western blotting, activity analysis of caspase-3, caspase-8 and caspase-9 and mitochondrial membrane potential measurement, the apoptosis induced by PEN-mediated p53 transfection was conducted in a mitochondria-dependent apoptosis pathway. These results demonstrated that PEN could successfully mediate the p53 gene delivery and up-regulate the cellular p53 expression level, triggering a significant p53-dependent anti-proliferative effect on tumor cells.
AuthorsJianxu Zhang, Di Wu, Zhen Xing, Shaojun Liang, Haobo Han, Hui Shi, Yan Zhang, Yan Yang, Quanshun Li
JournalColloids and surfaces. B, Biointerfaces (Colloids Surf B Biointerfaces) Vol. 129 Pg. 54-62 (May 01 2015) ISSN: 1873-4367 [Electronic] Netherlands
PMID25829127 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
CopyrightCopyright © 2015 Elsevier B.V. All rights reserved.
Chemical References
  • Acrylamides
  • TP53 protein, human
  • Tumor Suppressor Protein p53
  • Polyethyleneimine
  • N-isopropylacrylamide
  • Caspases
Topics
  • Acrylamides (chemistry)
  • Apoptosis
  • Blotting, Western
  • Caspases (metabolism)
  • Cell Cycle
  • Cell Proliferation
  • Cell Survival
  • Drug Delivery Systems
  • Gene Transfer Techniques
  • Genetic Therapy
  • HeLa Cells
  • Humans
  • Male
  • Membrane Potential, Mitochondrial
  • Mitochondria (metabolism)
  • Polyethyleneimine (chemistry)
  • Prostatic Neoplasms (genetics, pathology, therapy)
  • Signal Transduction (drug effects)
  • Transfection
  • Tumor Cells, Cultured
  • Tumor Suppressor Protein p53 (chemistry, genetics, metabolism)

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