1. Hepatocyte-targeting liposomal co-delivery of siRNA and Doxorubicin for enhanced anticancer efficacy

10. Liposome productionComponent.condition Cat No. D-gal. ceramide(c12) L Avanti 860544P PEG-DSPEL Avanti 880120P CholesterolL Sigma C3045-5G DMKEL- POPCL Avanti 850457P Rho-DOPEL Avanti 810150C Sonication

11. Encapsulation (siRNA) DOX

12. Transfection (receptor-mediated gene transfer) siRNA

13. Cell viability assay

14. Figure 1. Gel-retardation assay to determine optimal ratio of siRNA and Liposome generation. 1234 5 67 1.siRNA ( Control) 2.siRNA : Liposome = 1:20 3.siRNA : Liposome = 1:30 4.siRNA : Liposome = 1:40 5.siRNA : Liposome = 1:50 6.siRNA : Liposome = 1:60 7.siRNA : Liposome = 1:70 siRNA : Liposome = w:w

15. Figure 2. Gel-retardation assay to determine optimal ratio of siRNA and Liposome generation with SDS. abc de fg h i kj a.siRNA (Control) b.siRNA : Liposome = 1:20 c.siRNA : Liposome = 1:20 + SDS d.siRNA : Liposome = 1:40 e.siRNA : Liposome = 1:40 + SDS f.siRNA : Liposome = 1:60 g.siRNA : Liposome = 1:60 + SDS h.siRNA : Liposome = 1:70 i.siRNA : Liposome = 1:70 + SDS j.siRNA : Liposome = 1:80 k.siRNA : Liposome = 1:80 + SDS siRNA : Liposome = w:w

16. A549 Huh7 Figure 2. Cellular specific binding of Liposome to galactose-binding receptor in vitro. A549 siRNA-lipoplexes (siRNA + Liposome, Neutral complex ) Drug-liposome (Doxorubicin + Liposome, Positive complex) Huh7 A549Huh7

17. Dox-lipo (µl) Absorbance (450 nm) A549 Dox-lipo (µl) Absorbance (450 nm) Huh7 100% 91.10% 78% 60.99% 45.11% 39.01% Figure 3. Cytotoxicity of Dox-liposome on A549 and Huh7.

18. 1.Optimal ratio of siRNA and liposome is 1:70. 2.siRNA-liposome can bind to the hepatoma cell (Huh7) specifically. 3.DOX-liposome can bind to both lung adenocarcinoma (A549) and hepatoma cell. 4.Cytotoxicity of DOX-liposome is highly effective in hepatoma cell than lung adenocarcinoma cell.

19. 1.Find a optimal ratio of si-RNA, Doxorubicin and liposome. 2. Measure the decrement of expression level of vimentin using therapeutic vimentin-siRNA for cancer cell.