Figures & data
Figure 1. Effects of doxorubicin and cisplatin on the propagation of ZD55-IL-24.
(A) Quantitative PCR to detect the replication of ZD55-IL-24 in NCI-H460 cells.
(B) Analysis of IL-24 expression by Western blotting with the presence of doxorubicin and cisplatin. NCI-H460 cells were infected with ZD55-IL-24 at an MOI of 10 pfu/cell, and IL-24 protein was analyzed 48 hours later in cell lysates. A total of 20 μg protein was loaded on 10% polyacrylamide gels and separated. Lane 1 control, lane 2 ADM (80 nM), lane 3 DDP (200 nM), lane 4 ZD55-IL24, lane 5 ZD55-IL24+ADM (80 nM), lane 6 ZD55-IL24 + DDP (200 nM).
Figure 2. Cell killing effect of ZD55-IL-24 was enhanced when combined with Doxorubicin and cisplatin against cancer cell lines but not normal cells.
Cells were infected with 5 moi (A549 and HeLa), 10 moi (NCI-H460 and HT-1080), or 20 moi (L-02). Cell viability was measured with MTT assay 72 hours after treatment.
Figure 3. The combination induced much more tumor-specific apoptosis in NCI-H460 cells.
(A) Cell apoptotic staining. After 72 hrs of treatment, NCI-H460 cells were incubated with Hoechst 33342 for 30 minutes, and condensation and fragmentation of nuclei were observed under a fluorescence microscope (arrow).
(B) The cleavage of caspase substrate PARP was assessed. PKR expression was also determined. Lane 1 control, lane 2 ADM (80 nM), lane 3 DDP (200 nM), lane 4 ZD55-IL24, lane 5 ZD55-IL24+ADM (80 nM), lane 6 ZD55-IL24 + DDP (200 nM), lane 7 ZD55-EGFP, lane 8 ZD55-EGFP+ADM (80 nM), lane 9 ZD55-EGFP + DDP (200 nM).
Figure 4. Treatment of NCI-H460 xenograft with ZD55-IL-24 and adriamycin.
Tumor size was measured weekly after injection of ZD55-IL-24 (1 × 109 pfu/tumor) or ZD55-EGFP (1 × 109 pfu/tumor). Adriamycin was injected intraperitoneally (4 mg/kg) at the same time. Each group included at least five mice.
H&E staining of tumors obtained from mice treated with ZD55-IL-24 and/ or adriamycin. The tumor treated with PBS was used as control.
Original magnification ×400. a: PBS as control, b: ADM, c: ZD55-IL24, d: ZD55-IL24+ADM.
Immunohistochemical staining with anti-E1A was performed. Original magnification ×400. Original magnification ×400. a: PBS as control, b: ZD55-IL24, c: ZD55-IL24+ADM.
Apoptotic study in vivo by TUNEL after treatment with viruses. TUNEL assay was performed to detect apoptosis in the tumor section. Original magnification x400. a: PBS as control, b: ADM, c: ZD55-IL24, d: ZD55-IL24+ADM.
