Basic & Clinical Medicine PLGA MUC1 MUC1 IC 50 MUC1 +

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1 July 2011 Vol. 31 No. 7 MUC * MUC1 PLGA MUC1 MUC1 MUC1 MCF-7 HepG2 MTS IC 50 MUC ± 9. 2 nm 83. 6% ± 1. 7% MUC1 + MCF-7 P < IC mg /L MUC1 + IC 50 MUC1 R A Preparation of MUC1-targeted nanoparticles and evaluation of its cytotoxicity in vitro YU Chen-chen 1 HU Yan 1 DUAN Jin-hong 1 WANG Chen 2 XU Hai-yan 1 YANG Xian-da 1* 1. Dept. of Physiology and Pathophysiology Institute of Basic Medical Sciences CAMS & PUMC Beijing National Center of Nanoscience and Technology Beijing China Abstract Objective To construct MUC1-targeted nanoparticles and evaluate its cytotoxicity in vitro. Methods MUC1 aptamers were conjugated to the surface of nanoparticles which were made of poly lactic-co-glycolic-acid PLGA nanoparticles and loaded with paclitaxel PTX. The MUC1-targeted nanoparticles Apt-NP-PTX were characterized by UV spectrophotometry and dynamic light scattering. Using MUC1-overexpressing MCF-7 breast cancer cell as experimental group and HepG2 as control group the specificity of the Apt-NP-PTX was detected by flow cytometry. The cytotoxicity and IC 50 of Apt-NP-PTX against MCF-7 were evaluated using a standard MTS assay. Results The Apt-NP-PTX were about ± 9. 2 nm in size with an encapsulation efficiency of 83. 6% ± 1. 7%. The Apt-NP-PTX enhanced in vitro drug delivery and cellular toxicity to MUC1 + cancer cells as compared with non-targeted nanoparticles that lack the MUC1 aptamer P < with IC 50 of mg /L and mg /L respectively. Conclusion MCF-7 cells in vitro. Key words MUC1 aptamer targeted drug delivery tumor The Apt-NP-PTX can effectively enhance the PTX delivery to MUC1-overexpressing CB CB CB * corresponding author ayangmd@ gmail. com

2 aptamer 30 min DNA RNA MUC1 S g /L mol /L EDC mol /L NHS 15 min 3'-NH S h T - 30 min MUC NP PTX-NP 1. 0 mg 6 MUC1 S nm MUC1 PTX PTX poly lactidecoglycolide acid PLGA M w = ml PBS rpm EDC 5. 0 ml HCl N- NHS PBS MUC1 S2. 2 Invitrongen 188 M w = Rho-123 Sigma MUC1- MTS Promega MCF-7 MUC1 + HepG MCF h HepG2 S g /L mg PTX 2. 0 mg PLGA 0. 2 ml 1. 0 ml 5% min 1. 0 ml 1 mol /L NaOH MUC1-0. 5% 188 PBS ph mg /ml 5. 0 ml 3 Rho-123 MUC1 MUC1- MCF-7 HepG2 4 h D-Hank's % 1 ~ 2 min 4% 10 min MUC1-96 MCF-7 HepG h

3 MUC1 753 NP PTX PLGA 83. 6% ± 1. 7% NP-PTX Apt-NP-PTX 4. 2% ± 0. 1% MUC1 MTS MCF-7 D-Hank's μl RPMI μl MTS 37 1 h 490 nm A IC 50 MCF-7 HepG h PTX Fig 1 Structure of MUC1 aptamer conjugated NP-PTX Apt-NP-PTX 6 nanoparticles 4 h MUC1-96 MCF-7 MUC1 Apt-NP MTS S2. 2 IC % HepG2 ± x ± s Excel t 2 12 h 54. 1% % 3 MUC MUC ± 9. 2 nm PTX 4 NP MCF-7 HepG2 PLGA PTX 1 2 MUC1 Apt-PTX-NP 2. 3 MUC1-2 Fig 2 MCF-7 A HepG2 B NP Apt-NP Flow cytometry analysis of MCF-7 A and HepG2 B cells treated with Apt-NPs and NPs A. MCF-7 cells B. HepG2 cells

4 PTX NP-PTX Apt-NP-PTX IC mg /L Apt-NP-PTX PTX 8. 6 NP-PTX 1. 7 MCF-7 Apt-NP NP Fig 5 MTS PTX NP-PTX Apt-NP-PTX MUC1 + MCF-7 IC 50 MTS assay to determine the IC 50 of PTX NP-PTX and Apt-NP-PTX towards MCF-7 cells 3 Enhanced permeability and retention 8 MUC1 MUC1 MCF-7 S2. 2 Apt-NP-PTX P < MUC HepG2 MUC1 S2. 2 MUC1 S2. 2 MUC IC 50 MCF-7 MUC1 S PTX PTX MUC1 S2. 2 NP-PTX Apt-NP-PTX MCF-7 MCF-7 - HepG2 MUC1

5 MUC1 755 MUC1 IC 50 MUC1 PLGA MUC1 PTX MUC1 S2. 2 MUC1 S2. 2 S2. 2 MUC1 S2. 2 S2. 2 MUC1 + IC 50 MUC1 + MUC1 MUC1 MUC1 MUC1 MUC Levy-Nissenbaum E Radovic-Moreno AF Wang AZ et al. Nanotechnology and aptamers applications in drug delivery J. Trends Biotechnol Dhar S Gu FX Langer R et al. Targeted delivery of cisplatin to prostate cancer cells by aptamer functionalized Pt IV prodrug-plga-peg nanoparticles J. Proc Natl Acad Sci USA Farokhzad OC Jon S Khademhosseini A et al. Nanoparticle-aptamer bioconjugates a new approach for targeting prostate cancer cells J. Cancer Res Farokhzad OC Cheng J Teply BA et al. Targeted nanoparticle-aptamer bioconjugates for cancer chemotherapy in vivo J. Proc Natl Acad Sci USA Huang YF Shangguan D Liu H et al. Molecular assembly of an aptamer-drug conjugate for targeted drug delivery to tumor cells J. Chembiochem Taylor-Papadimitriou J Burchell J Miles DW et al. MUC1 and cancer J. BBA-Mol Basis Dis Ferreira CSM Matthews CS Missailidis S. DNA aptamers that bind to MUC1 tumour marker design and characterization of MUC1-binding single-stranded DNA aptamers J. Tumor Biol Yu B Tai HC Xue W et al. Receptor-targeted nanocarriers for therapeutic delivery to cancer J. Mol Membr Biol O'Connor JC Julian J Lim SD et al. MUC1 expression in human prostate cancer cell lines and primary tumors J. Prostate Cancer Prostatic Dis Ohuchida K Mizumoto K Yamada D et al. Quantitative analysis of MUC1 and MUC5AC mrna in pancreatic juice for preoperative diagnosis of pancreatic cancer J. Int J Cancer Chauhan SC Vinayek N Maher DM et al. Combined staining of TAG-72 MUC1 and CA125 improves labeling sensitivity in ovarian cancer antigens for multi-targeted antibody-guided therapy J. J Histochem Cytochem