Title:Melatonin Induces Apoptosis and Inhibits the Proliferation of Cancer Cells via Reactive Oxygen Species-mediated MAPK and mTOR Pathways
Volume: 7
Author(s): Reena Kasi, Pei Ling Yeo, Ng. Khuen Yen, Rhun Yian Koh, Gnanajothy Ponnudurai, Yee Lian Tiong and Soi Moi Chye*
Affiliation:
- School of Health Sciences, International Medical University, Kuala Lumpur 57000,Malaysia
Keywords:
Melatonin, apoptosis, cancer, reactive oxygen species, MAPK, mTOR.
Abstract:
Background: Recent human and animal studies have demonstrated the oncostatic properties
of N-acetyl-5-methoxytryptamine (melatonin) in different types of cancer. However, in few cancer
cell lines including colorectal cancer cell line (HT-29), acute T cell leukemia cell line (JURKAT)
and cervical cancer cell line (HeLa), precise oncostatic mechanism induced by melatonin is yet to be
described.
Objectives: The aim of this study is to investigate the effects of melatonin in HT-29, JURKAT and
HeLa cells and to determine the underlying molecular mechanism.
Methods: Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide
(MTT) assay while cell cycle, apoptosis and membrane potential were analysed by flow cytometry.
Reactive oxygen species (ROS) was detected by 2',7'-dichlorofluorescein diacetate (DCFH-DA)
staining. Protein expressions were determined by Western blot.
Results: Our results showed that melatonin suppressed cell proliferation, increased the number of sub
G1 hypodiploid cells and cell cycle arrest in HT-29, JURKAT and HeLa cells. Besides, melatonin also
induced early and late apoptosis, although there were marked variations in responses between different
cell lines (sensitivity; HeLa > HT-29 >JURKAT). Apart from that, staining with DCHF-DA
demonstrated ROS production that was induced in a dose-dependent manner in HeLa, HT-29 and
JURKAT cells. Moreover, the apoptotic process and oncostatic effect of melatonin were seen to be
associated with extracellular-signal-regulated kinase (ERK) and stress-activated protein kinase/c-Jun
NH (2)-terminal kinase (SAPK-JNK) signalling cascades in HeLa cells. In HT-29 and JURKAT cells,
melatonin induced apoptosis via activation of p38 mitogen-activated protein kinases (p38), ERK and
SAPK-JNK signalling pathways. In all three cell lines, the apoptotic event was triggered by the
mammalian target of rapamycin (mTOR)-mediated activation of the downstream target rapamycininsensitive
companion of mTOR (RICTOR) and/or regulatory-associated protein of mTOR (RAPTOR)
proteins.
Conclusion: Our findings confirm that melatonin induces apoptosis through reactive oxygen speciesmediated
dysregulated mitogen-activated protein kinase (MAPK) and mTOR signalling pathways in
these cancer cell lines.