Transfection of immortalized keratinocytes by low toxic poly(2-(dimethylamino)ethyl methacrylate)-based polymers.
Skin carcinoma are among the most spread diagnosed tumours in the world. In this study, we investigated the transfection of immortalized keratinocytes, used as an in vitro model for skin carcinoma, using antisense technology and poly(2-(dimethylamino)ethyl methacrylate) (PDMAEMA)-based polymers, with original architecture and functionalities. We tested PDMAEMA polymers with different structures: linear, with two (DEA-PDMAEMA) or three (TEA-PDMAEMA) arms. The cytotoxicity of these polymers was assessed over a wide range of apparent M n (from 7600 to 64 600). At a N/P ratio of 7.38, cytotoxicity increases with the M n. Keratinocytes were transfected with a fluorescent oligonucleotide and then analyzed by flow cytometry. For the three architectures tested, the percentage of transfected cells and abundance of internalized oligonucleotide were closely related to the M n of the polymer. Confocal microscopy and FACS analyses showed a wide spread fine granular distribution of the oligonucleotide up to 3 days post-transfection. Then, we assessed the silencing efficiency of the polymers, targeting GFP in GFP expressing keratinocytes. The maximal silencing effect (±40%) was obtained using a DEA-PDMAEMA polymer (M n = 30 300). These results suggest that PDMAEMA-based polymers can be efficiently used to transfect immortalized keratinocytes and, thus, open new perspectives in the therapy of skin carcinoma.
['Van Overstraeten-Schlögel|Nancy|N|', 'Ho-Shim|Yong|Y|', 'Tevel|Virginie|V|', 'Bontems|Sébastien|S|', 'Dubois|Philippe|P|', 'Raes|Martine|M|']
D049109:Cell Proliferation D005456:Fluorescent Dyes D020868:Gene Silencing D015316:Genetic Therapy D049452:Green Fluorescent Proteins D057809:HEK293 Cells D006576:Heterocyclic Compounds, 4 or More Rings D006801:Humans D015603:Keratinocytes D008689:Methacrylates D015394:Molecular Structure D009757:Nylons D016376:Oligonucleotides, Antisense D012333:RNA, Messenger D012878:Skin Neoplasms D014162:Transfection
TRANSFECTION KERATINOCYTES SKIN CARCINOMA 2-(DIMETHYLAMINO)ETHYL METHACRYLATE ANTISENSE OLIGONUCLEOTIDE HOSTASOL