MCF7 (PLGA) Zeta MCF7 85%,

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1 ( ; ; ; ) : [ ] (PLGA) MCF7 [ ] MCF7 Zeta ; [ ] MCF7 75nm MCF7 85% 3 (P<0.05) [ ] MCF7 MCF7 : ; ; ; :R737.9 :A : (2010) Preparation of Breast Cancer Cell Lysates-loaded Nanoparticle and Preliminary Evaluation of Its Immunity in vitro MA Jin 1 DUAN Jin-hong 1 ZHANG Qi-jun 2 et al. (1.Institute of Basic Medicine Chinese Academy of Medical Sciences and Peking Union Medical College Beijing China; 2.Technical Institute of Physics and Chemistry Chinese Academy of Science Beijing China) Abstract: [Purpose] To prepare nanoparticle for encapsulating MCF7 breast cancer cell lysate protein (MCF7p) with poly (D L-lactide co-glycolide) copolymer (PLGA) and to evaluate its role in immunotherapy for breast cancer. [Methods] The double emulsion method was employed for preparation of nanoparticles (NP) encapsulating the MCF7 breast cancer cell lysate protein. The morphology size zeta potential and encapsulation efficiency of the NP were assayed and optimized. The NP was utilized to simulate the peripheral blood mononuclear cells (PBMC) and the anti-tumor immune reaction of NP was detected in vitro. [Results] The obtained NPs were negatively charged spherical in shape relatively uniform in size with a mean diameter of 75nm and a protein encapsulation rate of 85%.The NP-stimulated PMBC generated a significantly higher anticancer cytotoxicity in vitro with 3 times compared to the control group(p<0.05). [Conclusion] The NPs prepared are of good physicochemical properties and can encapsulate protein antigen with high efficiency. Moreover the NPs significantly enhance the immunogenicity of the MCF7 tumor lysate protein suggesting that nanotechnology may have potential applications in immunotherapy against cancer. Key words: tumor lysate protein; nanoparticles; breast cancer; immunotherapy [2] : ; : [1] : (2006CB )China Medical Board : ayangmd@gmail.com China Cancer2010Vol.19No.6

2 ); ; [3] ; MCF NP MCF7 80% 0.25% ( NP [4] 0.01%EDTA) RPMI- (poly lactic-co-glycolic acidplga) 1640 : D-hanks (%)=( - )/ FDA 100% 1mol/L NaOH [6] cocktail( NP cocktail 100 PLGA 1) 4 2h : MCF r/min 15min (%)=NP / % (nanoparticlesnp) (W1/ [5] O/W2) MCF7 NPs MCF7 ( % 188 ) 1ml 25 (pro- PLGA tease inhibitor cocktail Zeta Sigma ); 5% 1~2 (PLGA g/mol ); 188( ); ( );Bradford 30min MTS BCA MCF7 NPs ( MCF7 NP NP );MTS( )( 96 ) (Promega );RPMI-1640 ( (PBMC) 4 ); (FBS : HyClone ); ( ); NP ( (T cells +NP China Cancer2010Vol.19No.6 401

3 ); MCF7 (T cells+mcf7 ); MCF7 (MCF-NP) (T (1) cells+mcf7-np ) 72h MCF7 0.1μg/ml MCF7 (polydispersity 96 MCF7 indexpdi) 0.202< nm ( 4h (1) 2)Zeta -22.7mV (3) E T( : 1 MCF7 ) MCF7 (180s ) 2 (120s ); 1 MTS ( (%) ) ( 21.67% ) MCF % ( 2) :PLGA =10mg; 1 D-hanks 3 100μl RPMI μl MTS h A490 : (80%) (%)=1-(A490 -A MCF7 )/(A490 -A490 ) 100% MCF7 1.3 SPSS 13.0 t (W1/O/W2) China Cancer2010Vol.19No.6

4 Zeta (mv) 3 MCF T cells+mcf7 T cells+np T cells+mcf7-np *:P<0.05 ;#:P<0.05T cells+mcf7 ;E T= MCF7 2.3 MCF7 (P<0.05) PLGA MCF7 PLGA NP FDA Tcells+NP PBMC [7] MCF7 MTS NP MCF7 MCF7 (PBMC) MCF7 4 T 30% ( cells+np 30nm~200nm PBMC) MCF %±7.98% ( PBMC MCF7 [4] 9.36%±5.92%) 10% PLGA (%) China Cancer2010Vol.19No.6 403

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