Retrovirally mediated RNA interference targeting the M2 subunit of ribonucleotide reductase: A novel therapeutic strategy in pancreatic cancer.

摘要: Abstract Background Ribonucleotide reductase M2 subunit (RRM2) overexpression enhances tumor chemoresistance and cellular invasiveness. We hypothesized that the RNA interference (RNAi) induced by retrovirally delivered small interfering (siRNA) would sensitize pancreatic adenocarcinoma cells to gemcitabine attenuate their invasive potential. Methods Stable suppression of RRM2 expression in PANC1, MIAPaCa2, BxPC3, Capan2 was exposure a novel replication-deficient retrovirus, engineered express RRM2-specific siRNA (psiRRM2), confirmed Western blot analysis. Single-base mismatch vector (psiControl) served as control. activity quantified, 50% inhibitory concentrations were calculated. TUNEL staining caspase profiling performed after exposure. Cellular invasiveness quantified Matrigel Boyden chamber. NF-κB matrix metalloproteinase-9 (MMP-9) measured. Results stably specifically suppressed psiRRM2, but not psiControl transfectants. psiRRM2 transfectants exhibited lower concentrations, increased gemcitabine-induced apoptosis, greater caspase-3 activation, relative Invasiveness attenuated transfectants, activity, MMP-9 expression, Conclusions gene silencing attenuates chemoresistance. Retroviral delivery can efficiently induce stable RNAi, allowing dissection function potentially representing new therapeutic modality.