NaCl induced high cationic hydroxyethylated cholesterol-based nanoparticle-mediated synthetic small interfering RNA transfer into prostate carcinoma PC-3 cells.
The RNA interference (RNAi) effect is an alternative technology to antisense DNA as an experimental method of down-regulating a specific target protein. Optimal gene therapy for tumors must deliver synthetic small interfering RNA (siRNA) to tumor cells with high efficiency and minimal toxicity. Previously, we reported that cationic nanoparticles composed of cholesteryl-3beta-carboxyamidoethylene-N-hydroxyethylamine and Tween 80 (NP-OH) could deliver plasmid DNA with high transfection efficiency. In this study, to apply NP-OH for siRNA transfection, we optimized the charge ratio (+/-) of NP-OH/siRNA and nanoplex-forming solution, and evaluated the transfection efficiency into PC-3 cells. Positively charged nanoplex prepared in the presence of sodium chloride exhibited efficient siRNA transfer. The distribution of siRNA after transfection was strongly detected both in the cytoplasm and nucleus. Furthermore, we demonstrated that NP-OH nanoplex of bcl-2 siRNA significantly inhibited tumor growth compared with control siRNA, and the efficacy was comparable to commercial products. The results of the experiments showed that NP-OH nanoparticles have potential as a viable vector candidate for synthetic siRNA delivery.
Hattori
Yoshiyuki
Y
Institute of Medicinal Chemistry, Hoshi University, Tokyo, Japan. yhattori@hoshi.ac.jp
Yoshizawa
Takashi
T
Koga
Kimiko
K
Maitani
Yoshie
Y
D000970:Antineoplastic Agents / Q000008:administration & dosage / Q000737:chemistry; D015153:Blotting, Western; D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects; D002784:Cholesterol / Q000008:administration & dosage / Q000031:analogs & derivatives* / Q000737:chemistry; D004273:DNA, Neoplasm / Q000235:genetics; D004563:Electrochemistry; D004591:Electrophoresis, Polyacrylamide Gel; D005434:Flow Cytometry; D019254:Genes, bcl-2 / Q000235:genetics; D005822:Genetic Vectors; D006801:Humans; D008156:Luciferases / Q000235:genetics; D018613:Microscopy, Confocal; D053758:Nanoparticles; D010957:Plasmids / Q000235:genetics; D011136:Polysorbates; D034741:RNA, Small Interfering / Q000008:administration & dosage* / Q000737:chemistry / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D012965:Sodium Chloride / Q000494:pharmacology*; D013501:Surface-Active Agents; D014162:Transfection
D000970:Antineoplastic Agents; D004273:DNA, Neoplasm; D011136:Polysorbates; D034741:RNA, Small Interfering; D013501:Surface-Active Agents; C106621:cholesteryl-3-carboxyamidoethylene-N-hydroxyethylamine; D012965:Sodium Chloride; D002784:Cholesterol; D008156:Luciferases