RT Journal Article
SR Electronic
T1 Late-breaking abstract: EGFR specific single chain antibody deliver siRNAs restore gefitinib sensitivity in resistant NSCLC cells
JF European Respiratory Journal
JO Eur Respir J
FD European Respiratory Society
SP 3287
VO 44
IS Suppl 58
A1 Lu, Yuan
A1 Liu, Li
A1 Zhang, Rui
A1 Ren, Xinling
YR 2014
UL http://erj.ersjournals.com/content/44/Suppl_58/3287.abstract
AB To overcome TKIs resistance, we developed scFv and s-9R, two human single chain antibodies against EGFR(fig.1A). Here, we showed s-9R could deliver FAM-siRNA into EGFR positve NSCLC cells(fig.1B). A549, H1993 and H1975 were either transfected with KRAS, MET or EGFR siRNA or co-incubated with scFv/s-9R/ indicated siRNA mixtures, following treated with gefitinib. Compared with the control groups, the target genes' expression level were down-regulated in the s-9R/siRNA mixture groups detected by RT-qPCR and western blot(fig.1C and D). In vivo, s-9R/siRNA mixtures restored gefitinib sensitivity in TKIs resistance cells measured by MTT(fig.1E).In vitro, the fusion proteins were gradually cleaned from normal tissues and enriched in the tumor till 48h assessed by Living image system(fig.2A). The co-administration of s-9R/siRNA and gefitinib treatment successfully suppressed the xenograft tumor progression (fig.2B) and prolonged the life span of nude mouse(fig.2C and D).Therefore, the study provides not only a new scFv derivative to deliver siRNA into EGFR-positive NSCLC cells but also a flexible method to overcome TKIs resistance.