Functional gene silencing mediated by chitosan/siRNA nanocomplexes.
Chitosan/siRNA nanoparticles to knock down FHL2 gene expression were reported in this work. The physicochemical properties such as particle size, surface charge, morphology and complex stability of chitosan nanoparticle-incorporated siRNA were evaluated. Nanoparticles which were formulated with chitosan/siRNA exhibited irregular, lamellar and dendritic structures with a hydrodynamic radius size of about 148 nm and net positive charges with zeta-potential value of 58.5 mV. The knockdown effect of the chitosan/siRNA nanoparticles on gene expression in FHL2 over-expressed human colorectal cancer Lovo cells was investigated. The result showed that FHL2 siRNA formulated within chitosan nanoparticles could knock down about 69.6% FHL2 gene expression, which is very similar to the 68.8% reduced gene expression when siRNA was transfected with liposome Lipofectamine. Western analysis further showed significant FHL-2 protein expression reduced by the chitosan/siRNA nanoparticles. The results also showed that blocking FHL2 expression by siRNA could also inhibit the growth and proliferation of human colorectal cancer Lovo cells. The current results demonstrated that chitosan-based siRNA nanoparticles were a very efficient delivery system for siRNA in vivo as previously reported.
Ji
A M
AM
Department of Pharmaceutical Science, Zhujiang Hospital, Southern Medical University, Guangzhou 510282, People's Republic of China.
Su
D
D
Che
O
O
Li
W S
WS
Sun
L
L
Zhang
Z Y
ZY
Yang
B
B
Xu
F
F
D015153:Blotting, Western; D045744:Cell Line, Tumor; D048271:Chitosan / Q000737:chemistry*; D004586:Electrophoresis; D020868:Gene Silencing / Q000187:drug effects*; D018398:Homeodomain Proteins / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D060850:LIM-Homeodomain Proteins; D018625:Microscopy, Atomic Force; D008855:Microscopy, Electron, Scanning; D009124:Muscle Proteins / Q000235:genetics / Q000378:metabolism*; D053758:Nanoparticles / Q000737:chemistry* / Q000648:ultrastructure; D049329:Nanostructures / Q000737:chemistry* / Q000648:ultrastructure; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000494:pharmacology*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014157:Transcription Factors / Q000235:genetics / Q000378:metabolism*
C105987:FHL2 protein, human; D018398:Homeodomain Proteins; D060850:LIM-Homeodomain Proteins; D009124:Muscle Proteins; D034741:RNA, Small Interfering; D014157:Transcription Factors; D048271:Chitosan