ORIGINAL PAPER Inhibition of autophagy enhances DENSpm-induced apoptosis in human colon cancer cells in a p53 independent manner Ajda Coker Gurkan 1 & Elif Damla Arisan 1 & Pinar Obakan Yerlikaya 1 & Halime Ilhan 1 & Narcin Palavan Unsal 1 Accepted: 20 December 2017 # International Society for Cellular Oncology 2018 Abstract Purpose One of the recently developed polyamine (PA) analogues, N 1 ,N 11 -diethylnorspermine (DENSpm), has been found to act as an apoptotic inducer in melanoma, breast, prostate and colon cancer cells. Also, its potential to induce autophagy has been established. Unfolded protein responses and starvation of amino acids are known to trigger autophagy. As yet, however, the molecular mechanism underlying PA deficiency-induced autophagy is not fully clarified. Here, we aimed to determine the apoptotic effect of DENSpm after autophagy inhibition by 3- methyladenine (3-MA) or siRNA-mediated Beclin-1 silencing in colon cancer cells. Methods The apoptotic effects of DENSpm after 3-MA treatment or Beclin-1 silencing were determined by PI and AnnexinV/PI staining in conjunction with flow cytometry. Intracellular PA levels were measured by HPLC, whereas autophagy and the expression profiles of PA key players were determined in HCT116, SW480 and HT29 colon cancer cells by Western blotting. Results We found that DENSpm-induced autophagy was inhibited by 3-MA treatment and Beclin-1 silencing, and that apoptotic cell death was increased by PA depletion and spermidine/spermine N 1 -acetyltransferase (SSAT) up- regulation. We also found that autophagy inhibition led to DENSpm-induced apoptosis through Atg5 down-regula- tion, p62 degradation and LC3 lipidation in both HCT116 and SW480 cells. p53 deficiency did not alter the response of the colon cancer cells to DENSpm-induced apoptotic cell death under autophagy suppression conditions. Conclusions From our results we conclude that DENSpm-induced apoptotic cell death is increased when autophagy is inhibited by 3-MA or Beclin-1 siRNA through PA depletion and PA catabolic activation in colon cancer cells, regardless p53 mutation status. Keywords Colon cancer . Apoptosis . Autophagy . Polyamine analogue . DENSpm Abbreviations AO Acridine Orange AZI Antizyme Inhibitor BENSpm N 1 ,N 11 -bis (ethyl) norspermine BH Bcl-2 homology CHENSpm N 1 -ethyl-N 11 -((cycloheptyl) methyl)-4,8- diazaundecane DiOC 6 3,3′-Dihexyloxacarbocyanine Iodide DENSpm N 1 ,N 11 -diethylnorspermine DMSO Dimethylsulfoxide GFP Green Fluorescence Protein MDC Monodansyl Cadaverin MTT 3–4,5-Dimethyl-2-thiazolyl-2,5-diphenyl-2H- tetrazolium bromide ODC Ornithine Decarboxylase PAO Polyamine Oxidase PBS Phosphate-buffered saline PI Propidium Iodide Put Putrescine PVDF Polyvinyldifluoride SDS-PAGE Sodium dodecyl sulphate polyacrylamide gel electrophoresis Spd Spermidine Spm Spermine SSAT Spermidine/spermine N 1 -acetyltransferase TBS Tris-buffered saline * Ajda Coker Gurkan a.coker@iku.edu.tr 1 Molecular Biology and Genetics Department, Science and Letters Faculty, Istanbul Kultur University, Atakoy Campus, 34156 Istanbul, Turkey Cellular Oncology https://doi.org/10.1007/s13402-017-0369-x