Naslov
Increased promoter activity is responsible for increased adenovirus transduction efficacy of human laryngeal carcinoma cells resistant to vincristine

Autori
Majhen, Dragomira ; Brozović, Anamaria ; Buger, Tvrtko ; Osmak, Maja ; Ambriović-Ristov, Andreja

Vrsta, podvrsta i kategorija rada
Sažeci sa skupova, sažetak, znanstveni

Izvornik
ESGCT 2008 Abstracts, XVIth Annual Congress of the European Society of Gene and Cell Therapy, Concertgebouw, Brugge, Belgium, November 13– 16, 2008 / - : Mary Ann Liebert, Inc., 2008, 1109-1109

Skup
XVIth Annual Congress of the European Society of Gene and Cell Therapy

Mjesto i datum
Brugge, Belgija, 13.11.2008. - 16.11.2008

Vrsta sudjelovanja
Poster

Vrsta recenzije
Nije recenziran

Ključne riječi
drug resistance; gene therapy; adenovirus

Sažetak
Adenoviral gene therapy is an approach for treating cancers resistant to currently available therapies such as drug resistant cells. Adenovirus infection occurs via binding of the fiber knob domain to CAR (coxsackie and adenovirus receptor), followed by endocytosis of the virion through interaction of penton base Arg-Gly-Asp (RGD) motif with cellular integrins. On the model of human laryngeal carcinoma (HEp2)-derived vincristine resistant cell line, we observed 3-fold increased wild type adenovirus (Ad5RSVβ gal) transduction efficacy in comparison with parental HEp2 cells, while for Ad5 639RSVβ gal with short fibers, which infects cells independent of CAR, this difference was 9-fold. We found decreased expression of CAR, increased expression of α vβ 3 integrin and equal amounts of α vβ 5 integrin in VK2 in comparison to HEp2 cells. In VK2 cells we found slightly smaller amount of internalized virus as well as decreased Ad5RSVβ gal attachment as compared to HEp2 cells. Finally, Ad5CMVβ gal, being essentially the same as Ad5wtRSVβ gal except for promoter driving expression of transgene, showed equal transduction efficacy in both HEp2 and VK2 cell lines. Therefore we hypothesized that this discrepancy between internalization/attachment and transduction efficacy is likely due to the different activity of RSV and CMV promoters in HEp2 and VK2 cells. We compared the activity of these promoters after transient transfection of plasmids pRSVCAT and pCMVCAT in HEp2 and VK2 cells. In VK2 cells the activity of the RSV promoter was approximately 8-fold higher than in HEp2 cells, while we observed approximately 2-fold increased activity of CMV promoter in VK2 in comparison to HEp2 cells. We conclude that increased promoter activity is responsible for increased adenovirus transduction efficacy of cells resistant to vincristine. Our results indicate that, when evaluating transduction of adenovirus vectors for gene therapy, not only cell surface CAR and/or integrin levels should be measured but also transgene expression driven by different promoters.

Izvorni jezik
Engleski

Znanstvena područja
Biologija

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