• 文献标题:   Nano-Graphene Oxide-supported APTES-Spermine, as Gene Delivery System, for Transfection of pEGFP-p53 into Breast Cancer Cell Lines
  • 文献类型:   Article
  • 作  者:   MIRZAIE V, ANSARI M, NEMATOLLAHIMAHANI SN, NASERY MM, KARIMI B, ESLAMINEJAD T, POURSHOJAEI Y
  • 作者关键词:   dna carrier, in vitro, gene transfer, conjugated polymer, mcf7 cell, nanomedicine
  • 出版物名称:   DRUG DESIGN DEVELOPMENT THERAPY
  • ISSN:   1177-8881
  • 通讯作者地址:   Kerman Univ Med Sci
  • 被引频次:   0
  • DOI:   10.2147/DDDT.S251005
  • 出版年:   2020

▎ 摘  要

Purpose: Genetic diseases can be the result of genetic dysfunctions that happen due to some inhibitory and/or environmental risk factors, which are mostly called mutations. One of the most promising treatments for these diseases is correcting the faulty gene. Gene delivery systems are an important issue in improving the gene therapy efficiency. Therefore, the main purpose of this study was modifying graphene oxide nanoparticles by spermine in order to optimize the gene delivery system. Methods: Graphene oxide/APTES was modified by spermine (GOAS) and characterized by FT-IR, DLS, SEM and AFM techniques. Then pEGFP-p53 was loaded on GOAS, transfected into cells and evaluated by fluorescent microscopy and gene expression techniques. Results: FT-IR data approved the GOAS sheet formation. Ninety percent of the particles were less than 56 nm based on DLS analysis. SEM analysis indicated that the sheets were dispersed with no aggregation. AFM results confirmed the dispersed structures with thickness of 1.25 +/- 0.87 nm. STA analysis showed that GOAS started to decompose from 400 degrees C and was very unstable during the heating process. The first weight loss up to 200 degrees C was due to the evaporation of absorbed water, the second one observed in the range of 200-550 degrees C was assigned to the decomposition of labile oxygen- and nitrogen-containing functional groups, and the third one above 550 degrees C was attributed to the removal of oxygen functionalities. In vitro release of DNA demonstrated the efficient activity of the new synthesized system. Ninety percent of the cells were transfected and showed the GFP under fluorescence microscopy, and TP53 gene was expressed 51-fold in BT-20 cells compared to beta-actin as the reference gene. Flow cytometry analysis confirmed the apoptosis of the cells rather than necrosis. Conclusion: It could be concluded that the new synthesized structure could transfer a high amount of the therapeutic agent into cells with best activity.