Salinomycin induces endoplasmic reticulum stress‑mediated autophagy and apoptosis through generation of reactive oxygen species in human glioma U87MG cells

Yu,Sun-Nyoung; Kim,Sang-Hun; Kim,Kwang-Youn; Ji,Jae-Hoon; Seo,Young-Kyo; Yu,Hak Sun; Ahn,Soon-Cheol

doi:10.3892/or.2017.5615

Salinomycin induces endoplasmic reticulum stress‑mediated autophagy and apoptosis through generation of reactive oxygen species in human glioma U87MG cells

Authors:
- Sun-Nyoung Yu
- Sang-Hun Kim
- Kwang-Youn Kim
- Jae-Hoon Ji
- Young-Kyo Seo
- Hak Sun Yu
- Soon-Cheol Ahn
View Affiliations

Affiliations: Department of Microbiology and Immunology, Pusan National University School of Medicine, Yangsan 50612, Republic of Korea, Department of Herbal Formula, Medical Research Center (MRC-GHF), College of Oriental Medicine, Daegu Haany University, Gyeongsan 38610, Republic of Korea, Genome Instability Research, Ajou University School of Medicine, Suwon 16502, Republic of Korea, School of Life Sciences, Ulsan National Institute of Science and Technology, Ulsan 44919, Republic of Korea, Immunoregulatory Therapeutics Group in Brain Busan 21 Project, Pusan National University, Yangsan 50612, Republic of Korea
Published online on: May 2, 2017 https://doi.org/10.3892/or.2017.5615
Pages: 3321-3328

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Salinomycin is a polyether ionophore antibiotic that has recently been shown to induce cell apoptosis in human cancer cells displaying multiple mechanisms of drug resistance. In the present study, we explored the impact of salinomycin on the apoptosis and autophagy as well as the correlation between those effects and endoplasmic reticulum (ER) stress molecular mechanisms in human glioma U87MG cells. Apoptosis, autophagy and reactive oxygen species (ROS) were analyzed using flow cytometry. In addition, expression levels of apoptosis-, autophagy- and ER stress-related proteins were determined by western blotting. The results showed that salinomycin induced apoptosis, ER stress and autophagy in glioma cancer cell lines. In addition, salinomycin also induced ROS generation, and the ROS scavenger N-acetyl-L-cysteine was found to inhibit the salinomycin-induced apoptosis, ER stress and autophagy. The inhibition of ER stress with 4-phenylbutyric acid depressed salinomycin-induced apoptosis and autophagy. Salinomycin increased the expression of autophagy marker protein, LC3B, and accumulation of acidic vesicular organelles. Furthermore, pre-treatment with the autophagy inhibitor 3-methyladenine showed potential in increasing the apoptosis rate induced by salinomycin in the U87MG cells. Taken together, these results revealed that salinomycin induced apoptosis and autophagy via ER stress mediated by ROS, suggesting that ER stress by salinomycin plays a dual function in both promoting and suppressing cell death.

View Figures

View References

1	Dong TT, Zhou HM, Wang LL, Feng B, Lv B and Zheng MH: Salinomycin selectively targets ‘CD133⁺’ cell subpopulations and decreases malignant traits in colorectal cancer lines. Ann Surg Oncol. 18:1797–1804. 2011. View Article : Google Scholar : PubMed/NCBI
2	Wang F, He L, Dai WQ, Xu YP, Wu D, Lin CL, Wu SM, Cheng P, Zhang Y, Shen M, et al: Salinomycin inhibits proliferation and induces apoptosis of human hepatocellular carcinoma cells in vitro and in vivo. PLoS One. 7:e506382012. View Article : Google Scholar : PubMed/NCBI
3	Kusunoki S, Kato K, Tabu K, Inagaki T, Okabe H, Kaneda H, Suga S, Terao Y, Taga T and Takeda S: The inhibitory effect of salinomycin on the proliferation, migration and invasion of human endometrial cancer stem-like cells. Gynecol Oncol. 129:598–605. 2013. View Article : Google Scholar : PubMed/NCBI
4	Kim KY, Yu SN, Lee SY, Chun SS, Choi YL, Park YM, Song CS, Chatterjee B and Ahn SC: Salinomycin-induced apoptosis of human prostate cancer cells due to accumulated reactive oxygen species and mitochondrial membrane depolarization. Biochem Biophys Res Commun. 413:80–86. 2011. View Article : Google Scholar : PubMed/NCBI
5	Kim SH, Choi YJ, Kim KY, Yu SN, Seo YK, Chun SS, Noh KT, Suh JT and Ahn SC: Salinomycin simultaneously induces apoptosis and autophagy through generation of reactive oxygen species in osteosarcoma U2OS cells. Biochem Biophys Res Commun. 473:607–613. 2016. View Article : Google Scholar : PubMed/NCBI
6	Xipell E, Gonzalez-Huarriz M, de Irujo JJ Martinez, García-Garzón A, Lang FF, Jiang H, Fueyo J, Gomez-Manzano C and Alonso MM: Salinomycin induced ROS results in abortive autophagy and leads to regulated necrosis in glioblastoma. Oncotarget. 7:30626–30641. 2016.PubMed/NCBI
7	Chu SH, Feng DF, Ma YB, Zhang H, Zhu ZA, Li ZQ and Jiang PC: Promoter methylation and downregulation of SLC22A18 are associated with the development and progression of human glioma. J Transl Med. 9:1562011. View Article : Google Scholar : PubMed/NCBI
8	Wei KC, Huang CY, Chen PY, Feng LY, Wu TW, Chen SM, Tsai HC, Lu YJ, Tsang NM, Tseng CK, et al: Evaluation of the prognostic value of CD44 in glioblastoma multiforme. Anticancer Res. 30:253–259. 2010.PubMed/NCBI
9	Ohgaki H and Kleihues P: Epidemiology and etiology of gliomas. Acta Neuropathol. 109:93–108. 2005. View Article : Google Scholar : PubMed/NCBI
10	Das A, Banik NL and Ray SK: N-(4-Hydroxyphenyl) retinamide induced both differentiation and apoptosis in human glioblastoma T98G and U87MG cells. Brain Res. 1227:207–215. 2008. View Article : Google Scholar : PubMed/NCBI
11	Powis G, Gasdaska JR, Gasdaska PY, Berggren M, Kirkpatrick DL, Engman L, Cotgreave IA, Angulo M and Baker A: Selenium and the thioredoxin redox system: Effects on cell growth and death. Oncol Res. 9:303–312. 1997.PubMed/NCBI
12	Sun Y and Rigas B: The thioredoxin system mediates redox-induced cell death in human colon cancer cells: Implications for the mechanism of action of anticancer agents. Cancer Res. 68:8269–8277. 2008. View Article : Google Scholar : PubMed/NCBI
13	Simon HU, Haj-Yehia A and Levi-Schaffer F: Role of reactive oxygen species (ROS) in apoptosis induction. Apoptosis. 5:415–418. 2000. View Article : Google Scholar : PubMed/NCBI
14	Rigoulet M, Yoboue ED and Devin A: Mitochondrial ROS generation and its regulation: Mechanisms involved in H₂O₂ signaling. Antioxid Redox Signal. 14:459–468. 2011. View Article : Google Scholar : PubMed/NCBI
15	Jang JH, Kim YJ, Kim H, Kim SC and Cho JH: Buforin IIb induces endoplasmic reticulum stress-mediated apoptosis in HeLa cells. Peptides. 69:144–149. 2015. View Article : Google Scholar : PubMed/NCBI
16	Ron D and Hubbard SR: How IRE1 reacts to ER stress. Cell. 132:24–26. 2008. View Article : Google Scholar : PubMed/NCBI
17	Kouroku Y, Fujita E, Tanida I, Ueno T, Isoai A, Kumagai H, Ogawa S, Kaufman RJ, Kominami E and Momoi T: ER stress (PERK/eIF2alpha phosphorylation) mediates the polyglutamine-induced LC3 conversion, an essential step for autophagy formation. Cell Death Differ. 14:230–239. 2007. View Article : Google Scholar : PubMed/NCBI
18	Szegezdi E, Logue SE, Gorman AM and Samali A: Mediators of endoplasmic reticulum stress-induced apoptosis. EMBO Rep. 7:880–885. 2006. View Article : Google Scholar : PubMed/NCBI
19	White E and DiPaola RS: The double-edged sword of autophagy modulation in cancer. Clin Cancer Res. 15:5308–5316. 2009. View Article : Google Scholar : PubMed/NCBI
20	Amaravadi RK, Yu D, Lum JJ, Bui T, Christophorou MA, Evan GI, Thomas-Tikhonenko A and Thompson CB: Autophagy inhibition enhances therapy-induced apoptosis in a Myc-induced model of lymphoma. J Clin Invest. 117:326–336. 2007. View Article : Google Scholar : PubMed/NCBI
21	Yoon MJ, Kang YJ, Kim IY, Kim EH, Lee JA, Lim JH, Kwon TK and Choi KS: Monensin, a polyether ionophore antibiotic, overcomes TRAIL resistance in glioma cells via endoplasmic reticulum stress, DR5 upregulation and c-FLIP downregulation. Carcinogenesis. 34:1918–1928. 2013. View Article : Google Scholar : PubMed/NCBI
22	Lockshin RA and Zakeri Z: Apoptosis, autophagy, and more. Int J Biochem Cell Biol. 36:2405–2419. 2004. View Article : Google Scholar : PubMed/NCBI
23	Paglin S, Hollister T, Delohery T, Hackett N, McMahill M, Sphicas E, Domingo D and Yahalom J: A novel response of cancer cells to radiation involves autophagy and formation of acidic vesicles. Cancer Res. 61:439–444. 2001.PubMed/NCBI
24	Liu M, Ma S, Liu M, Hou Y, Liang B, Su X and Liu X: Synergistic killing of lung cancer cells by cisplatin and radiation via autophagy and apoptosis. Oncol Lett. 7:1903–1910. 2014.PubMed/NCBI
25	Malhi H, Gores GJ and Lemasters JJ: Apoptosis and necrosis in the liver: A tale of two deaths? Hepatology. 43(Suppl 1): S31–S44. 2006. View Article : Google Scholar : PubMed/NCBI
26	Eisenberg-Lerner A, Bialik S, Simon HU and Kimchi A: Life and death partners: Apoptosis, autophagy and the cross-talk between them. Cell Death Differ. 16:966–975. 2009. View Article : Google Scholar : PubMed/NCBI
27	Maycotte P and Thorburn A: Autophagy and cancer therapy. Cancer Biol Ther. 11:127–137. 2011. View Article : Google Scholar : PubMed/NCBI
28	Delaunay-Moisan A and Appenzeller-Herzog C: The antioxidant machinery of the endoplasmic reticulum: Protection and signaling. Free Radic Biol Med. 83:341–351. 2015. View Article : Google Scholar : PubMed/NCBI
29	Cheng Z and Ristow M: Mitochondria and metabolic homeostasis. Antioxid Redox Signal. 19:240–242. 2013. View Article : Google Scholar : PubMed/NCBI
30	Wang CL, Liu C, Niu LL, Wang LR, Hou LH and Cao XH: Surfactin-induced apoptosis through ROS-ERS-Ca²⁺-ERK pathways in HepG2 cells. Cell Biochem Biophys. 67:1433–1439. 2013. View Article : Google Scholar : PubMed/NCBI
31	Moir RD, Gross DA, Silver DL and Willis IM: SCS3 and YFT2 link transcription of phospholipid biosynthetic genes to ER stress and the UPR. PLoS Genet. 8:e10028902012. View Article : Google Scholar : PubMed/NCBI