Loss of Mitochondrial DNA by Gemcitabine Triggers Mitophagy and Cell Death

• Published in: Biological & pharmaceutical bulletin 2019/12/01, Vol.42(12), pp.1977-1987
• Main Authors: Inamura, Akihiro, Muraoka-Hirayama, Sanae, Sakurai, Koichi
• Format: Article
• Language: English
• Subjects: 1-Phosphatidylinositol 3-kinase; AKT protein; Animals; Antimetabolites, Antineoplastic - pharmacology; Apoptosis; Autophagy; Autophagy-Related Protein 5 - metabolism; Autophagy-Related Protein 7 - metabolism; Beta cells; Cell death; Cell Death - drug effects; Cell Line, Tumor; Chemotherapy; Deoxycytidine - analogs & derivatives; Deoxycytidine - pharmacology; Deoxyribonucleic acid; DNA; DNA damage; DNA, Mitochondrial; Gemcitabine; Inhibitors; Insulinoma; Kinases; Localization; Membranes; Mitochondria - drug effects; Mitochondria - metabolism; Mitochondrial DNA; mitochondrial membrane potential; Mitophagy; Mitophagy - drug effects; Neuroendocrine tumors; Pancreas; Phagocytosis; Rats; Superoxide; Therapeutic applications
• ISSN: 0918-6158; 1347-5215
• DOI: 10.1248/bpb.b19-00312

Gemcitabine (2,2-difluorodeoxycytidine nucleic acid), an anticancer drug exhibiting a potent ability to kill cancer cells, is a frontline chemotherapy drug. Although some chemotherapeutic medicines are known to induce nuclear DNA damage, no investigation into mitochondrial DNA (mtDNA) damage currently exists. When we treated insulinoma pancreatic β-cells (line INS-1) with high mitochondrial activity with gemcitabine for 24 h, the mtDNA contents were decreased. Gemcitabine induced a decrease in the number of mitochondria and the average potential of mitochondrial membrane in the cell but increased the superoxide anion radical levels. We observed that treatment with gemcitabine to induce cell death accompanied by autophagy-related protein markers, Atg5 and Atg7; these were significantly prevented by the autophagy inhibitors. The localization of Atg5 co-occurred with the location of mitochondria with membranes having high potential and mitophagy in cells treated with gemcitabine. The occurrence of mitophagy was inhibited by the inhibitors of the phosphatidylinositol 3-kinase/Akt pathway. Our results led us to the conclusion that gemcitabine induced cell death through mitophagy with the loss of mtDNA. These findings may provide a rationale for the combination of mtDNA damage with mitophagy in future clinical applications for cancer cells.