Preparation of folic acid/chitosan-mediated gene survivin shRNA nano-composites***⋆

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Abstract

BACKGROUND:

Folic acid-chitosan nanoparticle is a new nano-high-targeted vector, which can further improve drug targeting as well as the achievement of sustained-release and controlled-release drug delivery.

OBJECTIVE:

To study the feasibility of folic acid-chitosan nanoparticles as a recombinant survivin shRNA plasmid vector delivery system, as well as the transfection efficiency to colon cancer cells (SW480).

DESIGN, TIME AND SETTING:

A contrast experiment was performed at the Key Laboratory of Nanobiotechnology, Ministry of Public Health, Central South University between June 2008 and January 2009.

MATERIALS:

Chitosan (deacetylating degree > 90%) was provided by Shanghai Bo'ao Biological Technology Co., Ltd., folic acid was provided by sinopharm Chemical Reagent Co., Ltd., and survivin shRNA recombinant plasmid was provided by Shanghai GeneChem Co., Ltd.

METHODS:

Folate-conjugated chitosan nanoparticles with the uniform diameter were prepared, and then survivin shRNA (20 mg/L) recombinant plasmid and folic acid-chitosan (10 mg/L) were mixed to obtain nanocomposites. Meanwhile, cationic liposome gene complexes were considered as a control. Both the above-mentioned samples were used to transfect colorectal cancer cells.

MAIN OUTCOME MEASURES:

Physico-chemical property and transfection efficiency of nanocomposites and survivin protein expression in tumor cells after transfection using Western blotting.

RESULTS:

Folic acid-chitosan-mediated survivin shRNA recombinant plasmid nanocomposites were successfully prepared. The transfection efficiency of the nanocomposite was superior to that of cationic liposome gene complex to transfect colorectal cancer cells. In addition, survivin protein expression in the transfected colorectal cancer cells was less than cationic liposome gene complex.

CONCLUSION:

Folic acid-chitosan nanoparticles as a vector system can high-effectively delivery survivin shRNA recombinant plasmid into colon cancer cells so as to induce the apoptosis of human colorectal cancer cells.

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