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Pro‐apoptotic gene knockdown mediated by nanocomplexed siRNA reduces radiation damage in primary salivary gland cultures
Author(s) -
Arany Szilvia,
Xu Qingfu,
Hernady Eric,
Benoit Danielle S.W.,
Dewhurst Steve,
Ovitt Catherine E.
Publication year - 2012
Publication title -
journal of cellular biochemistry
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.028
H-Index - 165
eISSN - 1097-4644
pISSN - 0730-2312
DOI - 10.1002/jcb.24064
Subject(s) - small interfering rna , gene knockdown , transfection , rna interference , microbiology and biotechnology , apoptosis , gene silencing , salivary gland , cell , biology , cytoplasm , submandibular gland , gene , rna , biochemistry , endocrinology
A critical issue in the management of head and neck tumors is radioprotection of the salivary glands. We have investigated whether siRNA‐mediated gene knock down of pro‐apoptotic mediators can reduce radiation‐induced cellular apoptosis in salivary gland cells in vitro. We used novel, pH‐responsive nanoparticles to deliver functionally active siRNAs into cultures of salivary gland cells. The nanoparticle molecules are comprised of cationic micelles that electrostatically interact with the siRNA, protecting it from nuclease attack, and also include pH‐responsive endosomolytic constituents that promote release of the siRNA into the target cell cytoplasm. Transfection controls with Cy3‐tagged siRNA/nanoparticle complexes showed efficiently internalized siRNAs in more than 70% of the submandibular gland cells. We found that introduction of siRNAs specifically targeting the Pkcδ or Bax genes significantly blocked the induction of these pro‐apoptotic proteins that normally occurs after radiation in cultured salivary gland cells. Furthermore, the level of cell death from subsequent radiation, as measured by caspase‐3, TUNEL, and mitochondrial disruption assays, was significantly decreased. Thus, we have successfully demonstrated that the siRNA/nanoparticle‐mediated knock down of pro‐apoptotic genes can prevent radiation‐induced damage in submandibular gland primary cell cultures. J. Cell. Biochem. 113: 1955–1965, 2012. © 2012 Wiley Periodicals, Inc.