Inhibition of autophagy enhances DENSpm-induced apoptosis in human colon cancer cells in a p53 independent manner.

Abstract

PURPOSE:

One of the recently developed polyamine (PA) analogues, N 1 ,N11-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 N1-acetyltransferase (SSAT) upregulation. We also found that autophagy inhibition led to DENSpm-induced apoptosis through Atg5 down-regulation, 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.

• Authors: Ajda Coker Gurkan, Elif Damla Arisan, Pinar Obakan Yerlikaya, Halime Ilhan, Narcin Palavan Unsal
• Journal: Cellular oncology (Dordrecht) (Cell Oncol (Dordr)) Vol. 41 Issue 3 Pg. 297-317 (Jun 2018) ISSN: 2211-3436 [Electronic] Netherlands
• PMID: 29492901 (Publication Type: Journal Article)

Chemical References

• Antineoplastic Agents
• Beclin-1
• Polyamines
• RNA, Small Interfering
• TP53 protein, human
• Tumor Suppressor Protein p53
• N(1),N(11)-diethylnorspermine
• Spermine
• 3-methyladenine
• Caspase 3
• Adenine

Topics

• Adenine (analogs & derivatives, pharmacology)
• Antineoplastic Agents (pharmacology, therapeutic use)
• Apoptosis (drug effects)
• Autophagy (drug effects)
• Beclin-1 (antagonists & inhibitors, genetics)
• Caspase 3 (metabolism)
• Cell Cycle Checkpoints (drug effects)
• Cell Line, Tumor
• Cell Survival (drug effects)
• Colonic Neoplasms (drug therapy, metabolism, physiopathology)
• HCT116 Cells
• HT29 Cells
• Humans
• Membrane Potential, Mitochondrial (drug effects)
• Polyamines (metabolism)
• RNA, Small Interfering (pharmacology)
• Spermine (analogs & derivatives, pharmacology, therapeutic use)
• Tumor Suppressor Protein p53 (physiology)