Alantolactone induces apoptosis in RKO cells through the generation of reactive oxygen species and the mitochondrial pathway

Zhang,Yu; Bao,Yong  Li; Wu,Yin; Yu,Chun  Lei; Huang,Yan  Xin; Sun,Ying; Zheng,Li  Hua; Li,Yu  Xin

doi:10.3892/mmr.2013.1640

Alantolactone induces apoptosis in RKO cells through the generation of reactive oxygen species and the mitochondrial pathway

Authors:
- Yu Zhang
- Yong Li Bao
- Yin Wu
- Chun Lei Yu
- Yan Xin Huang
- Ying Sun
- Li Hua Zheng
- Yu Xin Li
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Affiliations: National Engineering Laboratory for Druggable Gene and Protein Screening, Northeast Normal University, Changchun, Jilin 130024, P.R. China, Research Center of Agriculture and Medicine Gene Engineering of Ministry of Education, Normal University, Changchun, Jilin 130024, P.R. China, Institute of Genetics and Cytology, Northeast Normal University, Changchun, Jilin 130024, P.R. China
Published online on: August 16, 2013 https://doi.org/10.3892/mmr.2013.1640
Pages: 967-972

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Abstract

Alantolactone, a methanol extract of Inula helenium, possesses anticancer properties in a number of cancer cell lines. However, its anticancer effect on human colorectal cancer cells and the underlying mechanisms remain to be elucidated. In the present study, the effects of alantolactone on cell viability and apoptosis in RKO human colon cancer cells were investigated. Alantolactone treatment of RKO cells was found to result in dose‑dependent inhibition of cell viability and induction of apoptosis, accompanied with the accumulation of reactive oxygen species (ROS) and the disruption of mitochondrial membrane potential. In addition, these effects were blocked with N‑acetylcysteine, a specific ROS inhibitor. Western blotting indicated that exposure of RKO cells to alantolactone is associated with the downregulation of Bcl‑2, induction of Bax and activation of caspase‑3 and ‑9. These results indicated that a ROS‑mediated mitochondria‑dependent pathway is involved in alantolactone‑induced apoptosis. From these observations, it was hypothesized that alantolactone may be used for the treatment of human colon cancer.

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1	Jemal A, Murray T, Ward E, et al: Cancer statistics, 2005. CA Cancer J Clin. 55:10–30. 2005. View Article : Google Scholar
2	Gustin DM and Brenner DE: Chemoprevention of colon cancer: current status and future prospects. Cancer Metastasis Rev. 21:323–348. 2002. View Article : Google Scholar : PubMed/NCBI
3	Fauzi AN, Norazmi MN and Yaacob NS: Tualang honey induces apoptosis and disrupts the mitochondrial membrane potential of human breast and cervical cancer cell lines. Food Chem Toxicol. 49:871–878. 2011. View Article : Google Scholar : PubMed/NCBI
4	Hsu HF, Houng JY, Kuo CF, Tsao N and Wu YC: Glossogin, a novel phenylpropanoid from Glossogyne tenuifolia, induced apoptosis in A549 lung cancer cells. Food Chem Toxicol. 46:3785–3791. 2008.PubMed/NCBI
5	Luo M, Liu X, Zu Y, et al: Cajanol, a novel anticancer agent from Pigeonpea [Cajanus cajan (L.) Millsp] roots, induces apoptosis in human breast cancer cells through a ROS-mediated mitochondrial pathway. Chem Biol Interact. 188:151–160. 2010.PubMed/NCBI
6	Cantrell CL, Abate L, Fronczek FR, et al: Antimycobacterial eudesmanolides from Inula helenium and Rudbeckia subtomentosa. Planta Med. 65:351–355. 1999. View Article : Google Scholar
7	Konishi T, Shimada Y, Nagao T, Okabe H and Konoshima T: Antiproliferative sesquiterpene lactones from the roots of Inula helenium. Biol Pharm Bull. 25:1370–1372. 2002. View Article : Google Scholar : PubMed/NCBI
8	Dirsch VM, Stuppner H and Vollmar AM: Cytotoxic sesquiterpene lactones mediate their death-inducing effect in leukemia T cells by triggering apoptosis. Planta Med. 67:557–559. 2001. View Article : Google Scholar : PubMed/NCBI
9	Lawrence NJ, McGown AT, Nduka J, Hadfield JA and Pritchard RG: Cytotoxic Michael-type amine adducts of alpha-methylene lactones alantolactone and isoalantolactone. Bioorg Med Chem Lett. 11:429–431. 2001. View Article : Google Scholar : PubMed/NCBI
10	Shi Y, Bao YL, Wu Y, et al: Alantolactone inhibits cell proliferation by interrupting the interaction between Cripto-1 and activin receptor type II A in activin signaling pathway. J Biomol Screen. 16:525–535. 2011. View Article : Google Scholar : PubMed/NCBI
11	Bellamy CO, Malcomson RD, Harrison DJ and Wyllie AH: Cell death in health and disease: the biology and regulation of apoptosis. Semin Cancer Biol. 6:3–16. 1995. View Article : Google Scholar : PubMed/NCBI
12	Reed JC: Apoptosis-regulating proteins as targets for drug discovery. Trends Mol Med. 7:314–319. 2001. View Article : Google Scholar : PubMed/NCBI
13	Kaufmann SH, Kottke TJ, Martins LM, Henzing AJ and Earnshaw WC: Analysis of caspase activation during apoptosis. Curr Protoc Cell Biol. Chapter 18(Unit 18): 22001.PubMed/NCBI
14	Danial NN and Korsmeyer SJ: Cell death: critical control points. Cell. 116:205–219. 2004. View Article : Google Scholar : PubMed/NCBI
15	Murphy KM, Ranganathan V, Farnsworth ML, Kavallaris M and Lock RB: Bcl-2 inhibits Bax translocation from cytosol to mitochondria during drug-induced apoptosis of human tumor cells. Cell Death Differ. 7:102–111. 2000. View Article : Google Scholar : PubMed/NCBI
16	Chatterjee S, Kundu S and Bhattacharyya A: Mechanism of cadmium induced apoptosis in the immunocyte. Toxicol Lett. 177:83–89. 2008. View Article : Google Scholar
17	Pathak N and Khandelwal S: Role of oxidative stress and apoptosis in cadmium induced thymic atrophy and splenomegaly in mice. Toxicol Lett. 169:95–108. 2007. View Article : Google Scholar : PubMed/NCBI
18	Martindale JL and Holbrook NJ: Cellular response to oxidative stress: signaling for suicide and survival. J Cell Physiol. 192:1–15. 2002. View Article : Google Scholar : PubMed/NCBI
19	Zhou YJ, Zhang SP, Liu CW and Cai YQ: The protection of selenium on ROS mediated-apoptosis by mitochondria dysfunction in cadmium-induced LLC-PK(1) cells. Toxicol In Vitro. 23:288–294. 2009. View Article : Google Scholar : PubMed/NCBI
20	Yang JS, Chen GW, Hsia TC, et al: Diallyl disulfide induces apoptosis in human colon cancer cell line (COLO 205) through the induction of reactive oxygen species, endoplasmic reticulum stress, caspases casade and mitochondrial-dependent pathways. Food Chem Toxicol. 47:171–179. 2009. View Article : Google Scholar
21	Wei A, Zhou D, Xiong C, Cai Y and Ruan J: A novel non-aromatic B-ring flavonoid: isolation, structure elucidation and its induction of apoptosis in human colon HT-29 tumor cell via the reactive oxygen species-mitochondrial dysfunction and MAPK activation. Food Chem Toxicol. 49:2445–2452. 2011. View Article : Google Scholar
22	Wang L, Xu ML, Hu JH, Rasmussen SK and Wang MH: Codonopsis lanceolata extract induces G0/G1 arrest and apoptosis in human colon tumor HT-29 cells - involvement of ROS generation and polyamine depletion. Food Chem Toxicol. 49:149–154. 2011. View Article : Google Scholar
23	Zhou L, Bao YL, Zhang Y, et al: Knockdown of TSP50 inhibits cell proliferation and induces apoptosis in P19 cells. IUBMB Life. 62:825–832. 2010. View Article : Google Scholar : PubMed/NCBI
24	Zhang Y, Zhou L, Bao YL, et al: Butyrate induces cell apoptosis through activation of JNK MAP kinase pathway in human colon cancer RKO cells. Chem Biol Interact. 185:174–181. 2010. View Article : Google Scholar : PubMed/NCBI
25	Rizzuto R, Nakase H, Darras B, et al: A gene specifying subunit VIII of human cytochrome c oxidase is localized to chromosome 11 and is expressed in both muscle and non-muscle tissues. J Biol Chem. 264:10595–10600. 1989.PubMed/NCBI
26	Zafarullah M, Li WQ, Sylvester J and Ahmad M: Molecular mechanisms of N-acetylcysteine actions. Cell Mol Life Sci. 60:6–20. 2003. View Article : Google Scholar
27	Rego AC and Oliveira CR: Mitochondrial dysfunction and reactive oxygen species in excitotoxicity and apoptosis: implications for the pathogenesis of neurodegenerative diseases. Neurochem Res. 28:1563–1574. 2003. View Article : Google Scholar : PubMed/NCBI
28	Shen Z, Shen J, Chen M, Li Q and Hong C: Morphological changes of mitochondria in apoptosis of esophageal carcinoma cells induced by As(2)O(3). Zhonghua Bing Li Xue Za Zhi. 29:200–203. 2000.(In Chinese).
29	Gryfe R, Swallow C, Bapat B, et al: Molecular biology of colorectal cancer. Curr Probl Cancer. 21:233–300. 1997. View Article : Google Scholar
30	Sadowska AM, Manuel Y-Keenoy B and De Backer WA: Antioxidant and anti-inflammatory efficacy of NAC in the treatment of COPD: discordant in vitro and in vivo dose-effects: a review. Pulm Pharmacol Ther. 20:9–22. 2007. View Article : Google Scholar : PubMed/NCBI
31	Budihardjo I, Oliver H, Lutter M, Luo X and Wang X: Biochemical pathways of caspase activation during apoptosis. Annu Rev Cell Dev Biol. 15:269–290. 1999. View Article : Google Scholar : PubMed/NCBI
32	Kroemer G and Martin SJ: Caspase-independent cell death. Nat Med. 11:725–730. 2005. View Article : Google Scholar : PubMed/NCBI
33	Adams JM and Cory S: The Bcl-2 protein family: arbiters of cell survival. Science. 281:1322–1326. 1998. View Article : Google Scholar : PubMed/NCBI
34	Yin XM: Signal transduction mediated by Bid, a pro-death Bcl-2 family proteins, connects the death receptor and mitochondria apoptosis pathways. Cell Res. 10:161–167. 2000. View Article : Google Scholar : PubMed/NCBI
35	Nomura M, Shimizu S, Ito T, et al: Apoptotic cytosol facilitates Bax translocation to mitochondria that involves cytosolic factor regulated by Bcl-2. Cancer Res. 59:5542–5548. 1999.PubMed/NCBI