Melatonin induces apoptosis in cholangiocarcinoma cell lines by activating the reactive oxygen species-mediated mitochondrial pathway

Laothong,Umawadee; Hiraku,Yusuke; Oikawa,Shinji; Intuyod,Kitti; Murata,Mariko; Pinlaor,Somchai

doi:10.3892/or.2015.3738

Melatonin induces apoptosis in cholangiocarcinoma cell lines by activating the reactive oxygen species-mediated mitochondrial pathway

Authors:
- Umawadee Laothong
- Yusuke Hiraku
- Shinji Oikawa
- Kitti Intuyod
- Mariko Murata
- Somchai Pinlaor
View Affiliations

Affiliations: Department of Parasitology, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand, Department of Environmental and Molecular Medicine, Mie University Graduate School of Medicine, Mie 514‑8507, Japan, Liver Fluke and Cholangiocarcinoma Research Center, Faculty of Medicine, Khon Kaen University, Khon Kaen 40002, Thailand
Published online on: January 20, 2015 https://doi.org/10.3892/or.2015.3738
Pages: 1443-1449

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

We previously demonstrated that melatonin could be used as a chemopreventive agent for inhibiting cholangiocarcinoma (CCA) development in a hamster model. However, the cytotoxic activity of melatonin in cancer remains unclear. In the present study, we investigated the effect of melatonin on CCA cell lines. Human CCA cell lines (KKU-M055 and KKU-M214) were treated with melatonin at concentrations of 0.5, 1 and 2 mM for 48 h. Melatonin treatment exerted a cytotoxic effect on CCA cells by inhibiting CCA cell viability in a concentration-dependent manner. Treatment with melatonin, especially at 2 mM, increased intracellular reactive oxygen species (ROS) production and in turn led to increased oxidative DNA damage and 8-oxodG formation. Moreover, melatonin treatment enhanced the production of cytochrome c leading to apoptosis in a concentration-dependent manner, as indicated by increased expression of apoptosis-related proteins caspase-3 and caspase-7. In conclusion, melatonin acts as a pro-oxidant by activating ROS-dependent DNA damage and thus leading to the apoptosis of CCA cells.

View Figures

View References

1	Anderson CD, Pinson CW, Berlin J and Chari RS: Diagnosis and treatment of cholangiocarcinoma. Oncologist. 9:43–57. 2004. View Article : Google Scholar : PubMed/NCBI
2	Tyson GL and El-Serag HB: Risk factors for cholangiocarcinoma. Hepatology. 54:173–184. 2011. View Article : Google Scholar : PubMed/NCBI
3	IARC. A Review of Human Carcinogens: Opisthorchis viverrini and Clonorchis sinensis. IARC Monogr Eval Carcinog Risks Hum. 100B:341–370. 2012.
4	Gerl R and Vaux DL: Apoptosis in the development and treatment of cancer. Carcinogenesis. 26:263–270. 2005. View Article : Google Scholar
5	Kaufmann SH and Earnshaw WC: Induction of apoptosis by cancer chemotherapy. Exp Cell Res. 256:42–49. 2000. View Article : Google Scholar : PubMed/NCBI
6	Hannun YA: Apoptosis and the dilemma of cancer chemotherapy. Blood. 89:1845–1853. 1997.PubMed/NCBI
7	Hail N Jr and Lotan R: Cancer chemoprevention and mitochondria: targeting apoptosis in transformed cells via the disruption of mitochondrial bioenergetics/redox state. Mol Nutr Food Res. 53:49–67. 2009. View Article : Google Scholar
8	Turrens JF: Mitochondrial formation of reactive oxygen species. J Physiol. 552:335–344. 2003. View Article : Google Scholar : PubMed/NCBI
9	Green DR and Reed JC: Mitochondria and apoptosis. Science. 281:1309–1312. 1998. View Article : Google Scholar : PubMed/NCBI
10	Desagher S and Martinou JC: Mitochondria as the central control point of apoptosis. Trends Cell Biol. 10:369–377. 2000. View Article : Google Scholar : PubMed/NCBI
11	Kasibhatla S and Tseng B: Why target apoptosis in cancer treatment? Mol Cancer Ther. 2:573–580. 2003.PubMed/NCBI
12	Zhang HM and Zhang Y: Melatonin: a well-documented anti-oxidant with conditional pro-oxidant actions. J Pineal Res. 57:131–146. 2014. View Article : Google Scholar : PubMed/NCBI
13	Bartsch C, Bartsch H and Karasek M: Melatonin in clinical oncology. Neuro Endocrinol Lett. 23:30–38. 2002.PubMed/NCBI
14	Jung B and Ahmad N: Melatonin in cancer management: progress and promise. Cancer Res. 66:9789–9793. 2006. View Article : Google Scholar : PubMed/NCBI
15	Laothong U, Pinlaor P, Boonsiri P, et al: Melatonin inhibits cholangiocarcinoma and reduces liver injury in Opisthorchis viverrini-infected and N-nitrosodimethylamine-treated hamsters. J Pineal Res. 55:257–266. 2013. View Article : Google Scholar : PubMed/NCBI
16	Sripa B, Leungwattanawanit S, Nitta T, et al: Establishment and characterization of an opisthorchiasis-associated cholangiocarcinoma cell line (KKU-100). World J Gastroenterol. 11:3392–3397. 2005. View Article : Google Scholar : PubMed/NCBI
17	Sanchez-Barcelo EJ, Cos S, Fernandez R and Mediavilla MD: Melatonin and mammary cancer: a short review. Endocr Relat Cancer. 10:153–159. 2003. View Article : Google Scholar : PubMed/NCBI
18	Hill SM, Frasch T, Xiang S, Yuan L, Duplessis T and Mao L: Molecular mechanisms of melatonin anticancer effects. Integr Cancer Ther. 8:337–346. 2009. View Article : Google Scholar
19	Barrenetxe J, Delagrange P and Martinez JA: Physiological and metabolic functions of melatonin. J Physiol Biochem. 60:61–72. 2004. View Article : Google Scholar : PubMed/NCBI
20	Bejarano I, Espino J, Barriga C, Reiter RJ, Pariente JA and Rodriguez AB: Pro-oxidant effect of melatonin in tumour leucocytes: relation with its cytotoxic and pro-apoptotic effects. Basic Clin Pharmacol Toxicol. 108:14–20. 2011. View Article : Google Scholar
21	Rodriguez C, Martin V, Herrera F, et al: Mechanisms involved in the pro-apoptotic effect of melatonin in cancer cells. Int J Mol Sci. 14:6597–6613. 2013. View Article : Google Scholar : PubMed/NCBI
22	Carbajo-Pescador S, Garcia-Palomo A, Martin-Renedo J, Piva M, Gonzalez-Gallego J and Mauriz JL: Melatonin modulation of intracellular signaling pathways in hepatocarcinoma HepG2 cell line: role of the MT1 receptor. J Pineal Res. 51:463–471. 2011. View Article : Google Scholar : PubMed/NCBI
23	Han Y, Demorrow S, Invernizzi P, et al: Melatonin exerts by an autocrine loop antiproliferative effects in cholangiocarcinoma: its synthesis is reduced favoring cholangiocarcinoma growth. Am J Physiol Gastrointest Liver Physiol. 301:G623–G633. 2011. View Article : Google Scholar : PubMed/NCBI
24	Motilva V, Garcia-Maurino S, Talero E and Illanes M: New paradigms in chronic intestinal inflammation and colon cancer: role of melatonin. J Pineal Res. 51:44–60. 2011. View Article : Google Scholar : PubMed/NCBI
25	Proietti S, Cucina A, Reiter RJ and Bizzarri M: Molecular mechanisms of melatonin’s inhibitory actions on breast cancers. Cell Mol Life Sci. 70:2139–2157. 2013. View Article : Google Scholar
26	Laothong U, Pinlaor P, Hiraku Y, et al: Protective effect of melatonin against Opisthorchis viverrini-induced oxidative and nitrosative DNA damage and liver injury in hamsters. J Pineal Res. 49:271–282. 2010. View Article : Google Scholar : PubMed/NCBI
27	Buyukavci M, Ozdemir O, Buck S, Stout M, Ravindranath Y and Savasan S: Melatonin cytotoxicity in human leukemia cells: relation with its pro-oxidant effect. Fundam Clin Pharmacol. 20:73–79. 2006. View Article : Google Scholar : PubMed/NCBI
28	Bejarano I, Espino J, Marchena AM, et al: Melatonin enhances hydrogen peroxide-induced apoptosis in human promyelocytic leukaemia HL-60 cells. Mol Cell Biochem. 353:167–176. 2011. View Article : Google Scholar : PubMed/NCBI
29	Lissoni P, Paolorossi F, Tancini G, et al: A phase II study of tamoxifen plus melatonin in metastatic solid tumour patients. Br J Cancer. 74:1466–1468. 1996. View Article : Google Scholar : PubMed/NCBI
30	Martin-Renedo J, Mauriz JL, Jorquera F, Ruiz-Andres O, Gonzalez P and Gonzalez-Gallego J: Melatonin induces cell cycle arrest and apoptosis in hepatocarcinoma HepG2 cell line. J Pineal Res. 45:532–540. 2008. View Article : Google Scholar : PubMed/NCBI
31	Osseni RA, Rat P, Bogdan A, Warnet JM and Touitou Y: Evidence of prooxidant and antioxidant action of melatonin on human liver cell line HepG2. Life Sci. 68:387–399. 2000. View Article : Google Scholar
32	Kim W, Jeong JW and Kim JE: CCAR2 deficiency augments genotoxic stress-induced apoptosis in the presence of melatonin in non-small cell lung cancer cells. Tumour Biol. 135:10919–10929. 2014. View Article : Google Scholar