title: The PEI-introduced CS shell/PMMA core nanoparticle for silencing the expression of E6/E7 oncogenes in human cervical cells.
abstract: In this study, we examined the potential of cationic nanoparticle - polyethyleneimine-introduced chitosan shell/poly (methyl methacrylate) core nanoparticles (CS-PEI) for siRNA delivery. Initially, DNA delivery was performed to validate the capability of CS-PEI for gene delivery in the human cervical cancer cell line, SiHa. siRNA delivery were subsequently carried out to evaluate the silencing effect on targeted E6 and E7 oncogenes. Physicochemical properties including size, zeta potential and morphology of CS-PEI/DNA and CS-PEI/siRNA complexes, were analyzed. The surface charges and sizes of the complexes were observed at different N/P ratios. The hydrodynamic sizes of the CS-PEI/DNA and CS-PEI/siRNA were approximately 300-400 and 400-500nm, respectively. Complexes were positively charged depending on the amount of added CS-PEI. AFM images revealed the mono-dispersed and spherical shapes of the complexes. Gel retardation assay confirmed that CS-PEI nanoparticles completely formed complexes with DNA and siRNA at a N/P ratio of 1.6. For DNA transfection, CS-PEI provided the highest transfection result. Localization of siRNA delivered through CS-PEI was confirmed by differential interference contrast (DIC) confocal imaging. The silencing effect of siRNA specific to HPV 16 E6/E7 oncogene was examined at 18 and 24h post-transfection. The results demonstrated the capacity of CS-PEI to suppress the expression of HVP oncogenes.
authors.affiliation: National Nanotechnology Center, National Science and Technology Development Agency, Pathumthani, Thailand. nattika@nanotec.or.th
authors.forename: Nattika
authors.initials: N
authors.lastname: Saengkrit
authors.forename: Phakorn
authors.initials: P
authors.lastname: Sanitrum
authors.forename: Noppawan
authors.initials: N
authors.lastname: Woramongkolchai
authors.forename: Somsak
authors.initials: S
authors.lastname: Saesoo
authors.forename: Nuttaporn
authors.initials: N
authors.lastname: Pimpha
authors.forename: Saowaluk
authors.initials: S
authors.lastname: Chaleawlert-Umpon
authors.forename: Tewin
authors.initials: T
authors.lastname: Tencomnao
authors.forename: Satit
authors.initials: S
authors.lastname: Puttipipatkhachorn
mesh_terms: D045744:Cell Line, Tumor; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects / Q000235:genetics; D015967:Gene Expression Regulation, Viral / Q000187:drug effects / Q000235:genetics; D020868:Gene Silencing; D052162:Human papillomavirus 16 / Q000235:genetics / Q000378:metabolism*; D006801:Humans; D020366:Methylmethacrylate / Q000737:chemistry* / Q000494:pharmacology; D053758:Nanoparticles / Q000737:chemistry*; D009856:Oncogene Proteins, Viral / Q000096:biosynthesis* / Q000235:genetics; D050725:Papillomavirus E7 Proteins / Q000096:biosynthesis* / Q000235:genetics; D030361:Papillomavirus Infections / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000628:therapy / Q000821:virology; D010316:Particle Size; D011094:Polyethyleneimine / Q000737:chemistry* / Q000494:pharmacology; D034741:RNA, Small Interfering / Q000737:chemistry* / Q000494:pharmacology; D012097:Repressor Proteins / Q000096:biosynthesis* / Q000235:genetics; D002583:Uterine Cervical Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000473:pathology / Q000628:therapy / Q000821:virology
chemical_list: C068651:E6 protein, Human papillomavirus type 16; D009856:Oncogene Proteins, Viral; D050725:Papillomavirus E7 Proteins; D034741:RNA, Small Interfering; D012097:Repressor Proteins; C059731:oncogene protein E7, Human papillomavirus type 16; D020366:Methylmethacrylate; D011094:Polyethyleneimine