Figure 1.

Reovirus preferentially replicates in KRas-transfected immortalized normal human pancreatic epithelial cells. (a) KRas transfected HPNE cells. Immunoblotting demonstrates KRas levels in HPNE cells. (b) Reovirus replicates preferentially in HPNE-KRas cells. Cells were treated with Reolysin for 48 h and stained with an anti-reovirus antibody. Immunocytochemistry reveals reovirus replication in KRas-transfected cells. Fluorescent intensity was quantified in HPNE and HPNE-KRas cells using Image-Pro Plus software version 6.2.1. Mean±S.D., n=5. *Indicates a significant difference compared with HPNE-vector cells. (c) KRas-transfected cells display higher levels of ER stress-related gene expression that can be further induced with reovirus exposure. HPNE-vector and HPNE-KRas cells were treated with 100 plaque-forming units (PFU)/cell Reolysin for 48 h. Gene expression was determined by qRT-PCR. Mean±S.D., n=3. ^#Represents a significant difference compared with vector control cells. *Indicates a significant difference compared with corresponding controls. (d) HPNE-KRas cells are sensitive to Reolysin-mediated cell death. Cells were treated for 72 h with the indicated concentrations of Reolysin, and cell viability was determined by MTT assay (left panel). Cells were treated for 48 h with Reolysin, and apoptosis was measured by PI-FACS analysis (right panel). Mean±S.D., n=3. *Indicates a significant difference compared with HPNE-vector cells treated with the same concentration of Reolysin P<0.05