A biomimetic nanovector-mediated targeted cholesterol-conjugated siRNA delivery for tumor gene therapy

被引：124

作者：

Ding, Yang ^{[1
]}

Wang, Wei ^{[1
]}

Feng, Meiqing ^{[2
]}

Wang, Yu ^{[3
]}

Zhou, Jianping ^{[1
]}

Ding, Xuefang ^{[1
]}

Zhou, Xin ^{[1
]}

Liu, Congyan ^{[1
]}

Wang, Ruoning ^{[1
]}

Zhang, Qiang ^{[4
]}

机构：

[1] China Pharmaceut Univ, Dept Pharmaceut, State Key Lab Nat Med, Nanjing 210009, Peoples R China

[2] Fu Dan Univ, Sch Pharm, Shanghai 201203, Peoples R China

[3] Nanjing Med Univ, Dept Pharmacol, Nanjing 210029, Jiangsu, Peoples R China

[4] Peking Univ, Sch Pharmaceut Sci, State Key Lab Nat & Biomimet Drugs, Beijing 100191, Peoples R China

来源：

BIOMATERIALS | 2012年 / 33卷 / 34期

基金：

中国国家自然科学基金;

关键词：

Reconstituted high density lipoprotein; Apolipoprotein A-I; Targeted cholesterol-conjugated siRNA delivery; Pokemon; Tumor gene therapy; HIGH-DENSITY-LIPOPROTEIN; ANTI-HBV ACTIVITY; IN-VIVO DELIVERY; CELL LUNG-CANCER; SR-BI; REPRESSES TRANSCRIPTION; SCAVENGER RECEPTOR; RNA INTERFERENCE; BREAST-CANCER; LIVER-CELLS;

D O I：

10.1016/j.biomaterials.2012.08.057

中图分类号：

R318 [生物医学工程];

学科分类号：

0831 ;

摘要：

RNA interference holds tremendous potential as a therapeutic approach of malignant tumors. However, safe and efficient nanovectors are extremely lack for systemic delivery of small interfering RNA (siRNA). The study aimed to develop a biomimetic nanovector, reconstituted high density lipoprotein (rHDL), mediating targeted cholesterol-conjugated siRNA (Chol-siRNA) delivery for Pokemon gene silencing therapy. Chol-siRNA-loaded rHDL nanoparticles (rHDL/Chol-siRNA complexes) were prepared using thin-film dispersion method and their characteristics were investigated in detail. RHDL/Chol-siRNA complexes at the optimal volume ratio (lipid: Chol-siRNA) exhibited high Chol-siRNA-loading efficiency (similar to 99%), desirable nanoparticle size and excellent stability in serum. In addition, by analyzing Chol-siRNA release profile, rHDL/Chol-siRNA complexes displayed sustained-release characteristic and storage stability. Observations from FACS and confocal microscopic analyses revealed that rHDL-mediated carboxyfluorescein tagged Chol-siRNA (FAM-Chol-siRNA) transfection resulted in highly efficient uptake and specific cytoplasmic delivery of FAM-Chol-siRNA into human hepatocellular carcinoma cell line HepG2 via HDL-receptor mediated mechanism. In vitro cytotoxicity, apoptosis and Western-blot analyses revealed significant cellular growth inhibition and decrease of Pokemon and Bcl-2 protein expression in HepG2 cells treated with Chol-siRNA-Pokemon-loaded rHDL nanoparticles (rHDL/Chol-siRNA-Pokemon complexes), respectively. In in vivo studies, the near-infrared (NIR) dye Cy5 labeled Chol-siRNA-loaded rHDL nanoparticles (rHDL/Cy5-Chol-siRNA complexes) obviously accumulated in tumor of nude mice after i.v. administration as compared with Cy5-Chol-siRNA-loaded lipoplexes (Lipos/Cy5-Chol-siRNA complexes). Morover, rHDL/Chol-siRNA-Pokemon complexes demonstrated great tumor growth inhibition and significant decrease of Pokemon and Bcl-2 protein expression in vivo. These results suggested that rHDL should be an ideal non-viral tumor-targeting vector for Chol-siRNA transfer, and rHDL-mediated Chol-siRNA-Pokemon delivery might be a promising new strategy for gene therapy in hepatocellular carcinoma. (C) 2012 Elsevier Ltd. All rights reserved.

引用

页码：8893 / 8905

页数：13

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