Abstract BACKGROUND: We have shown previously that inhibition of JNK1 renders HT29 colon adenocarcinoma cells more sensitive to hypoxia and/or oxaliplatin due, in part, to impaired autophagy induction. We also showed that the autophagy inhibitor chloroquine sensitizes HT29 to oxaliplatin in vitro and in mouse xenograft model. PURPOSE: Here we investigate further by molecular approaches, including modulation of signaling through JNK, how inhibition of autophagy affects sensitivity of hypoxic colon cancer cells to oxaliplatin. We employed a panel of HT29-derived cell lines: lines stably expressing dominant negative constructs for either JNK1 (HTJ1.3) or JNK2 (HTJ2.2) singly, derivatives of these lines in which the other JNK gene is silenced by viral delivery of shRNA construct (HTJ1s2 and HTJ2s1), as well as HT29-derived lines with down-regulated BclX (HTBX) or Beclin-1 (HTB). RESULTS: We found that down-regulation of JNK1 leads to inhibition of autophagy accompanied by increase in apoptotic (versus necrosis) cell death. Clonogenic assays revealed higher resistance of HTJ1s2 cells to oxaliplatin under hypoxia, whereas HTJ2s1 cells demonstrated slight increase in sensitivity to the drug in this setting. Down-regulation of either Beclin-1 or BclX resulted in significant sensitization to oxaliplatin in oxic and, even more so, in hypoxic conditions (three-folds on average), while exerting differing effects on autophagy induction. CONCLUSIONS: Our data demonstrate that JNK1 plays an important role in autophagy induction under hypoxic conditions in the HT29 colon adenocarcinoma cell line and that its inhibition increases apoptotic cell death in this setting. Inhibition of autophagy induction by down-regulation of JNK1 and knock-down of Beclin-1 or BclX resulted in reversal of oxaliplatin resistance in hypoxic HT29 cells. These findings suggest that targeting these proteins may be of value in colon cancer treatment. Citation Format: {Authors}. {Abstract title} [abstract]. In: Proceedings of the 103rd Annual Meeting of the American Association for Cancer Research; 2012 Mar 31-Apr 4; Chicago, IL. Philadelphia (PA): AACR; Cancer Res 2012;72(8 Suppl):Abstract nr 2267. doi:1538-7445.AM2012-2267
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