Piperlongumine induces G2/M phase arrest and apoptosis in cholangiocarcinoma cells through the ROS-JNK-ERK signaling pathway
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Cholangiocarcinoma (CCA) is an aggressive, metastatic bile duct cancer. CCA is difficult to diagnose, and responds poorly to current radio- and chemo-therapy. Piperlongumine (PL) is a naturally-occurring small molecule selectively toxic to cancer cells by targeting reactive oxygen species (ROS). In this study, we demonstrated the potential anticancer activity of PL in CCA. PL markedly induced death in CCA cell lines in a dose- and time-dependent manner through the activation of caspase-3 and PARP. PL also stimulated ROS accumulation in CCA. Co-exposure of PL with the ROS scavenger N-acetyl-l-cysteine or GSH completely blocked PL-induced apoptosis in CCA cell lines. Increased p21 via the p53-independent pathway in PL-treated CCA cells led to G2/M phase arrest and cell apoptosis. In addition, the study showed that PL trigger CCA cell lines death through JNK-ERK activation. Furthermore, the different antioxidant capacity of CCA cell lines also indicates the susceptibility of the cells to PL treatment. Our findings reveal that PL exhibits anti-tumor activity and has potential to be used as a chemotherapeutic agent against CCA.
KeywordsCholangiocarcinoma Piperlongumine Anti-tumor activity Reactive oxygen species Cell apoptosis
A three letter name (not an abbreviation); “p” stands for the phosphorylated (active) form
B cell lymphoma gene 2
BCL-2-associated protein X
BCL-2-associated death promoter
Phosphorylated (active) extracellular-signal-regulated kinase
Phosphorylated (active) C-Jun NH2-terminal kinase
Poly (ADP-ribose) polymerase
21 kDa cyclin-dependent kinase inhibitor
53 kDa tumor suppressor protein
Revolutions per minute
The authors thank (a) Professor Banchob Sripa of the Department of Pathology, Faculty of Medicine, Khon Kaen University for providing the cholangiocarcinoma cells; (b) the Biochemistry Laboratory, the Center for Scientific and Technological Equipment, Suranaree University of Technology for providing the research facilities; (c) Professor Jong-Lyel Roh of Department of Otolaryngology, Asan Medical Center, University of Ulsan College of Medicine, Seoul, Korea for providing some of the piperlongumine; and, (d) Mr. Bryan Roderick Hamman for assistance with the English-language presentation of the manuscript.
This work was supported by grants from Suranaree University of Technology (Grant Number: SUT1-102-57-24-23) and the Young Research Grants, Thailand Research Fund (MRG6080055) to CS. HC was supported by Grants (NRF-2015K1A4A3046807 and 2008-0062286) from the National Research Foundation of Korea and a Grant (2017-307) from the Asan Institute for Life Sciences, Seoul, Korea.
ST carried out all the experiments, performed data analysis, and prepared the manuscript draft. WS was provided advice on the cytotoxicity tests and data analysis. KJL supported and provided advice on the western blot analysis. HC was a grant holder, conceived the research, provided guidance on data analysis, revised and approved the manuscript. CT was a grant holder, designed the experiment, provided advice on the cell cycle analysis, ROS measurement, apoptosis assay, gene expression, and data analysis, revised and approved the manuscript.
Compliance with ethical standards
Conflict of interest
The authors declare no conflicts of interest.
- 1.Kamangar F, Dores GM, Anderson WF (2006) Patterns of cancer incidence, mortality, and prevalence across five continents: defining priorities to reduce cancer disparities in different geographic regions of the world. J Clin Oncol 24(14):2137–2150. doi: 10.1200/JCO.2005.05.2308 CrossRefPubMedGoogle Scholar
- 8.Raj L, Ide T, Gurkar AU, Foley M, Schenone M, Li X, Tolliday NJ, Golub TR, Carr SA, Shamji AF, Stern AM, Mandinova A, Schreiber SL, Lee SW (2011) Selective killing of cancer cells by a small molecule targeting the stress response to ROS. Nature 475(7355):231–234. doi: 10.1038/nature10167 CrossRefPubMedPubMedCentralGoogle Scholar
- 9.Xiong XX, Liu JM, Qiu XY, Pan F, Yu SB, Chen XQ (2015) Piperlongumine induces apoptotic and autophagic death of the primary myeloid leukemia cells from patients via activation of ROS-p38/JNK pathways. Acta Pharmacol Sin 36(3):362–374. doi: 10.1038/aps.2014.141 CrossRefPubMedPubMedCentralGoogle Scholar
- 13.Lee H-N, Jin H-O, Park J-A, Kim J-H, Kim J-Y, Kim B, Kim W, Hong S-E, Lee Y-H, Chang YH, Hong S-I, Hong YJ, Park I-C, Surh Y-J, Lee JK (2015) Heme oxygenase-1 determines the differential response of breast cancer and normal cells to piperlongumine. Mol Cell Biol 38(4):327–335. doi: 10.14348/molcells.2015.2235 Google Scholar
- 14.Duan C, Zhang B, Deng C, Cao Y, Zhou F, Wu L, Chen M, Shen S, Xu G, Zhang S, Duan G, Yan H, Zou X (2016) Piperlongumine induces gastric cancer cell apoptosis and G2/M cell cycle arrest both in vitro and in vivo. Tumour biol 37(8):10793–10804. doi: 10.1007/s13277-016-4792-9 CrossRefPubMedGoogle Scholar
- 17.Thongsom S, Chaocharoen W, Silsirivanit A, Wongkham S, Sripa B, Choe H, Suginta W, Talabnin C (2016) YKL-40/chitinase-3-like protein 1 is associated with poor prognosis and promotes cell growth and migration of cholangiocarcinoma. Tumour biol 37(7):9451–9463. doi: 10.1007/s13277-016-4838-z CrossRefPubMedGoogle Scholar
- 18.Jin HO, Lee YH, Park JA, Lee HN, Kim JH, Kim JY, Kim B, Hong SE, Kim HA, Kim EK, Noh WC, Kim JI, Chang YH, Hong SI, Hong YJ, Park IC, Lee JK (2014) Piperlongumine induces cell death through ROS-mediated CHOP activation and potentiates TRAIL-induced cell death in breast cancer cells. J Cancer Res Clin Oncol 140(12):2039–2046. doi: 10.1007/s00432-014-1777-1 CrossRefPubMedGoogle Scholar
- 21.Majewski N, Nogueira V, Bhaskar P, Coy PE, Skeen JE, Gottlob K, Chandel NS, Thompson CB, Robey RB, Hay N (2004) Hexokinase-mitochondria interaction mediated by Akt is required to inhibit apoptosis in the presence or absence of Bax and Bak. Mol Cell 16(5):819–830. doi: 10.1016/j.molcel.2004.11.014 CrossRefPubMedGoogle Scholar
- 26.Maruyama M, Kobayashi N, Westerman KA, Sakaguchi M, Allain JE, Totsugawa T, Okitsu T, Fukazawa T, Weber A, Stolz DB, Leboulch P, Tanaka N (2004) Establishment of a highly differentiated immortalized human cholangiocyte cell line with SV40T and hTERT. Transplantation 77(3):446–451. doi: 10.1097/01.TP.0000110292.73873.25 CrossRefPubMedGoogle Scholar
- 35.Chung JY, Hong SM, Choi BY, Cho H, Yu E, Hewitt SM (2009) The expression of phospho-AKT, phospho-mTOR, and PTEN in extrahepatic cholangiocarcinoma. Clin Cancer Res (2):660–667. doi: 10.1158/1078-0432.ccr-08-1084
- 37.Kongpetch S, Puapairoj A, Ong CK, Senggunprai L, Prawan A, Kukongviriyapan U, Chan-On W, Siew EY, Khuntikeo N, Teh BT, Kukongviriyapan V (2016) Haem oxygenase 1 expression is associated with prognosis in cholangiocarcinoma patients and with drug sensitivity in xenografted mice. Cell Prolif 49(1):90–101. doi: 10.1111/cpr.12228 CrossRefPubMedGoogle Scholar