![[Feedback]](/icons/banner/feedbackACT.gif){kind=icon}
![[For Subscribers]](/icons/banner/subscriberACT.gif){kind=icon}
![[Archive]](/icons/banner/archiveACT.gif){kind=icon}
![[Search]](/icons/banner/searchACT.gif){kind=icon}
![[Contents]](/icons/banner/tocACT.gif){kind=icon}
|
|
|
|
|
||
WY Gao, C Storm, W Egan and YC Cheng
Department of Pharmacology, Yale University School of Medicine, New Haven, Connecticut 06510.
The uptake and distribution of phosphorothioate oligodeoxynucleotides by human cells were studied using 35S-labeled 28-mer phosphorothioate oligodeoxycytidine [S-(dC)28]. Accumulation of intracellular S-(dC)28 was found to be higher in the carcinoma cells (grown in monolayers) than in the leukemia cells (grown in suspension culture). A hepatoma cell line transfected with hepatitis B virus, 2215, was chosen for further studies. The uptake of S-(dC)28 was partially dependent on temperature and energy. The intracellular concentration was significantly higher than that in the medium and the amount accumulated was dependent on the extracellular concentration. It appears that the uptake of S-(dC)28 involves mechanisms of both fluid-phase pinocytosis and adsorptive endocytosis. Neither oligonucleotides nor 5'- phosphorylated nucleotides inhibited S-(dC)28 uptake. Unlike horseradish peroxidase, which was primarily associated with endosomes once it was taken into the cell, S-(dC)28 was found to be present in both nuclear and cytoplasmic fractions. Efflux of S-(dC)28 from the cell was multiphasic; a trapping mechanism that could be due to a potent interaction of S-(dC)28 with cellular proteins was implicated. This trapping mechanism could be responsible for the lack of biological activity such as cytotoxicity and antisense activity of phosphorothioate oligodeoxynucleotides in some human cells.
This article has been cited by other articles:
|
|
 |
|
  S. M. Thomas, M. J. Ogagan, M. L. Freilino, S. Strychor, D. R. Walsh, W. E. Gooding, J. R. Grandis, and W. C. Zamboni Antitumor Mechanisms of Systemically Administered Epidermal Growth Factor Receptor Antisense Oligonucleotides in Combination with Docetaxel in Squamous Cell Carcinoma of the Head and Neck Mol. Pharmacol., March 1, 2008; 73(3): 627 - 638. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 S. H. Britz-Cunningham and S. J. Adelstein Molecular Targeting with Radionuclides: State of the Science J. Nucl. Med., December 1, 2003; 44(12): 1945 - 1961. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 P. de Diesbach, F. N'Kuli, M. Delmee, and P. J. Courtoy Infection by Mycoplasma hyorhinis strongly enhances uptake of antisense oligonucleotides: a reassessment of receptor-mediated endocytosis in the HepG2 cell line Nucleic Acids Res., February 1, 2003; 31(3): 886 - 892. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 D. P. Sester, S. Naik, S. J. Beasley, D. A. Hume, and K. J. Stacey Phosphorothioate Backbone Modification Modulates Macrophage Activation by CpG DNA J. Immunol., October 15, 2000; 165(8): 4165 - 4173. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 E. M. McGuffie, D. Pacheco, G. M. R. Carbone, and C. V. Catapano Antigene and Antiproliferative Effects of a c-myc-targeting Phosphorothioate Triple Helix-forming Oligonucleotide in Human Leukemia Cells Cancer Res., July 1, 2000; 60(14): 3790 - 3799. [Abstract] [Full Text]
|
|
|
|
|
|
C. Pichon, M. B. Roufai, M. Monsigny, and P. Midoux Histidylated oligolysines increase the transmembrane passage and the biological activity of antisense oligonucleotides Nucleic Acids Res., January 15, 2000; 28(2): 504 - 512. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 A. M. Gewirtz, D. L. Sokol, and M. Z. Ratajczak Nucleic Acid Therapeutics: State of the Art and Future Prospects Blood, August 1, 1998; 92(3): 712 - 736. [Full Text] [PDF]
|
|
|
|
|
|
D. G. Spiller, R. V. Giles, J. Grzybowski, D. M. Tidd, and R. E. Clark Improving the Intracellular Delivery and Molecular Efficacy of Antisense Oligonucleotides in Chronic Myeloid Leukemia Cells: A Comparison of Streptolysin-O Permeabilization, Electroporation, and Lipophilic Conjugation Blood, June 15, 1998; 91(12): 4738 - 4746. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 B. Hanss, E. Leal-Pinto, L. A. Bruggeman, T. D. Copeland, and P. E. Klotman Identification and characterization of a cell membrane nucleic acid channel PNAS, February 17, 1998; 95(4): 1921 - 1926. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 J. Lai, T. J. Crook, A. Payne, R. M. Lynch, and F. Porreca Antisense Targeting of Delta Opioid Receptors in NG 108-15 Cells: Direct Correlation between Oligodeoxynucleotide Uptake and Receptor Density J. Pharmacol. Exp. Ther., April 1, 1997; 281(1): 589 - 596. [Abstract] [Full Text]
|
|
|
|
 |
|
 C. Le Roy, P. Leduque, P. M. Dubois, J. M. Saez, and D. Langlois Repression of Transforming Growth Factor beta1 Protein by Antisense Oligonucleotide-induced Increase of Adrenal Cell Differentiated Functions J. Biol. Chem., May 3, 1996; 271(18): 11027 - 11033. [Abstract] [Full Text] [PDF]
|
|
|
|
 |
|
 M. R. Bennett and S. M. Schwartz Antisense Therapy for Angioplasty Restenosis : Some Critical Considerations Circulation, October 1, 1995; 92(7): 1981 - 1993. [Full Text]
|
|
|
|
 |
|
 C. Stein and Y. Cheng Antisense oligonucleotides as therapeutic agents--is the bullet really magical? Science, August 20, 1993; 261(5124): 1004 - 1012. [Abstract] [PDF]
|
|
 |
 |
 |
|
 |
 |
 |
