Autophagy is induced by adenoviral-mediated interferon a treatment in interferon resistant bladder cancer and normal urothelial cells as a cell death protective mechanism but not by the bystander factors produced

We have previously shown that adenoviral-mediated interferon alpha (Ad-IFN alpha) treatment is highly cytotoxic to tumor cells which are resistant to the IFN alpha protein. We now report that autophagy is produced after Ad-IFNa treatment of either IFN resistant bladder cancer cells (UC9 and KU7) or the normal urothelial cell line (TERT-NHUC). After Ad-IFNa infection autophagosomes, an early stage of autophagy, were seen in cancer cells whereas autophagolysosomes, a later stage of autophagy, were observed mostly in normal cells by electron microscopy. Conditioned medium from either normal or bladder cancer cells obtained after Ad-IFNa infection, however, produced no autophagy when placed on the bladder cancer cells, although again marked cytotoxicity was observed. This indicated that the autophagy seen was related to the direct effect of Ad-IFNa transfection and expression rather than to the bystander factors produced. In addition, autophagic changes were seen using LysoTracker Red DND-99 in both normal and cancer cells. We also documented that Ad-IFNa treatment produces the autophagic protein form, light chain 3 (LC3)-II, in cancer cells but not normal cells, which in turn was inhibited by the autophagic inhibitor, 3-methyladenine (3-MA). This inhibition of autophagy resulted in a significant increase in apoptotic cell death as measured by the sub-G1 population. We hypothesize that the autophagy seen in normal urothelial cells is a protective response and is allowed to be completed, providing a survival mechanism after Ad-IFN treatment, whereas the autophagy produced in IFN resistant cancer cells is not allowed to be completed and is insufficient to significantly suppress cytotoxicity. Cancer Gene Therapy (2010) 17, 579-584; doi:10.1038/cgt.2010.14; published online 21 May 2010