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Chitosan coated poly(lactic acid) polymeric nanoparticle-mediated combinatorial delivery of cisplatin and siRNA/plasmid DNA chemosensitizes cisplatin-resistant human ovarian cancer cells. Anish Babu, Qi Wang, Ranganayaki Muralidharan, Manish Shanker, Anupama Munshi, and Rajagopal Ramesh Mol. Pharmaceutics, Just Accepted Manuscript • Publication Date (Web): 12 Jun 2014 Downloaded from http://pubs.acs.org on June 19, 2014
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Molecular Pharmaceutics
ABSTRACT GRAPHIC
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Molecular Pharmaceutics
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Chitosan coated poly(lactic acid) polymeric nanoparticle-mediated combinatorial delivery of cisplatin and siRNA/plasmid DNA chemosensitizes cisplatin-resistant human ovarian cancer cells.
Anish Babu,1,2 Qi Wang,1,2 Ranganayaki Muralidharan,1,2 Manish Shanker,3# Anupama Munshi,2,4 Rajagopal Ramesh1,2,5*
1
Department of Pathology, 2Peggy and Charles Stephenson Cancer Center, University of Oklahoma
Health Sciences Center, Oklahoma City, Oklahoma 73104, USA; 3Department of Thoracic and Cardiovascular Surgery, The University of Texas MD Anderson Cancer Center, Houston, TX 77030, USA; 4Department of Radiation Oncology, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA; 5Graduate Program in Biomedical Sciences, University of Oklahoma Health Sciences Center, Oklahoma City, Oklahoma 73104, USA.
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Current address: University of Texas Dental School at Houston, Houston, TX 77030
*Address of Correspondence: Rajagopal Ramesh, Department of Pathology, The Biomedical Research Center, Suite 1403, 975 N.E., 10th Street, Oklahoma City, OK 73104, USA; Phone: (405) 271-6101; Email:
[email protected] 1 ACS Paragon Plus Environment
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Molecular Pharmaceutics
ABSTRACT Development of resistance towards anticancer drugs result in ineffective therapy leading to increased mortality. Therefore, overriding resistance and restoring sensitivity to anticancer drugs will improve treatment efficacy and reduce mortality. While numerous mechanisms for drug resistance in cancer have previously been demonstrated, recent studies implicate a role for proteasome and the autophagy regulatory protein P62/SQSTM1 (P62) in contributing to drug resistance. Specifically, reduction in the expression of the β5 subunit of the proteasome and/ or enhanced P62 protein expression is known to contribute to cancer drug resistance such as cisplatin (CDDP) in ovarian cancer cells. Therefore, we hypothesized that restoration of β5 expression and/or suppression of P62 protein expression in CDDP-resistant ovarian cancer cells will lead to restoration of sensitivity to CDDP and enhanced cell killing. To test our hypothesis we developed a biodegradable multifunctional nanoparticle (MNP) system that co-delivered p62siRNA, β5 plasmid DNA and CDDP and tested its efficacy in CDDP resistant 2008/C13 ovarian cancer cells. MNP consisted of CDDP loaded polylactic acid nanoparticle as inner core and cationic chitosan (CS) consisting of ionically linked P62siRNA (siP62) and/or β5expressing plasmid DNA (pβ5) as the outer layer. The MNPs were spherical in shape with a hydrodynamic diameter in the range of 280-350 nm, and demonstrated encapsulation efficiencies of 82% and 78.5% for CDDP and siRNA respectively. MNPs efficiently protected the siRNA and showed superior serum stability compared to naked siRNA as measured by gel retardation and spectrophotometry assays. The MNPs successfully delivered siP62 and pβ5 to cause P62 knockdown and restoration of β5 expression in 2008/C13 cells. Combined delivery of siP62, pβ5 and CDDP using the MNPs resulted in a marked reduction in the IC50 value of CDDP in 2008/C13 cells from 125 ± 1.3 µM to 98 ± 0.6 µM (P
