Oleuropein induces apoptosis via activation of caspases and suppression of phosphatidylinositol 3-kinase/protein kinase B pathway in HepG2 human hepatoma cell line

Yan,Chun‑Mei; Chai,Er‑Qing; Cai,Hong‑Yi; Miao,Guo‑Ying; Ma,Wen

doi:10.3892/mmr.2015.3266

Oleuropein induces apoptosis via activation of caspases and suppression of phosphatidylinositol 3-kinase/protein kinase B pathway in HepG2 human hepatoma cell line

Authors:
- Chun‑Mei Yan
- Er‑Qing Chai
- Hong‑Yi Cai
- Guo‑Ying Miao
- Wen Ma
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Affiliations: Department of Radiotherapy Oncology, Gansu Provincial Hospital, Lanzhou, Gansu 730000, P.R. China, Cerebrovascular Diseases Center, Gansu Provincial Hospital, Lanzhou, Gansu 730000, P.R. China
Published online on: January 28, 2015 https://doi.org/10.3892/mmr.2015.3266
Pages: 4617-4624

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Abstract

Oleuropein is a polyphenol, that is found in extra‑virgin olive oil. Previous studies have shown that oleuropein inhibits cell proliferation and induces apoptosis in breast cancer, colorectal cancer and thyroid cancer. The aim of the present study was to investigate the effects of oleuropein in hepatocellular carcinoma (HCC) cells. The results of Cell Counting Kit 8 and flow cytometric analysis indicated that oleuropein effectively inhibited cell viability and induced apoptosis in HepG2 human hepatoma cells in a dose‑dependent manner, through activation of the caspase pathway. Proapoptotic Bcl‑2 family members, BAX and Bcl‑2, were involved in oleuropein‑induced apoptosis. The phosphatidylinositol 3‑kinase/protein kinase B (PI3K/AKT) signaling pathway was also shown to be involved in this process. Oleuropein was demonstrated to suppress the expression of activated AKT. In addition, AKT overexpression promoted cell survival following treatment with oleuropein, while inhibition of AKT promoted cell death. Furthermore, the data demonstrated that oleuropein induces the production of reactive oxygen species (ROS) and that the function of oleuropein is, at least partially, ROS‑dependent. These results suggest that oleuropein may be a promising novel chemotherapeutic agent in hepatocellular carcinoma.

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1	Llovet JM, Burroughs A and Bruix J: Hepatocellular carcinoma. Lancet. 362:1907–1917. 2003. View Article : Google Scholar : PubMed/NCBI
2	Singh S, Singh PP, Roberts LR and Sanchez W: Chemopreventive strategies in hepatocellular carcinoma. Nat Rev Gastroenterol Hepatol. 11:45–54. 2013. View Article : Google Scholar : PubMed/NCBI
3	Cheng JW and Lv Y: New progress of non-surgical treatments for hepatocellular carcinoma. Med Oncol. 30:3812013. View Article : Google Scholar : PubMed/NCBI
4	Bruix J and Sherman M; American Association for the Study of Liver Diseases: Management of hepatocellular carcinoma: an update. Hepatology. 53:1020–1022. 2011. View Article : Google Scholar : PubMed/NCBI
5	Bravi F, Bosetti C, Tavani A, Gallus S and La Vecchia C: Coffee reduces risk for hepatocellular carcinoma: an updated meta-analysis. Clin Gastroenterol Hepatol. 11:1143–1421. 2013. View Article : Google Scholar
6	Calvisi DF, Ladu S, Hironaka K, Factor VM and Thorgeirsson SS: Vitamin E down-modulates iNOS and NADPH oxidase in c-Myc/TGF-alpha transgenic mouse model of liver cancer. J Hepatol. 41:815–822. 2004. View Article : Google Scholar : PubMed/NCBI
7	Sawada N, Inoue M, Iwasaki M, et al: Consumption of n-3 fatty acids and fish reduces risk of hepatocellular carcinoma. Gastroenterology. 142:1468–1475. 2012. View Article : Google Scholar : PubMed/NCBI
8	Hursting SD, Thornquist M and Henderson MM: Types of dietary fat and the incidence of cancer at five sites. Prev Med. 19:242–253. 1990. View Article : Google Scholar : PubMed/NCBI
9	Liu K, Stamler J, Moss D, Garside D, Persky V and Soltero I: Dietary cholesterol, fat, and fibre, and colon-cancer mortality. An analysis of international data. Lancet. 2:782–785. 1979. View Article : Google Scholar : PubMed/NCBI
10	Ferro-Luzzi A and Sette S: The Mediterranean Diet: an attempt to define its present and past composition. Eur J Clin Nutr. 43(Suppl 2): 13–29. 1989.PubMed/NCBI
11	Andreadou I, Mikros E, Ioannidis K, et al: Oleuropein prevents doxorubicin-induced cardiomyopathy interfering with signaling molecules and cardiomyocyte metabolism. J Mol Cell Cardiol. 69:4–16. 2014. View Article : Google Scholar : PubMed/NCBI
12	Hassan ZK, Elamin MH, Omer SA, et al: Oleuropein induces apoptosis via the p53 pathway in breast cancer cells. Asian Pac J Cancer Prev. 14:6739–6742. 2013. View Article : Google Scholar
13	Bulotta S, Corradino R, Celano M, et al: Antioxidant and antigrowth action of peracetylated oleuropein in thyroid cancer cells. J Mol Endocrinol. 51:181–189. 2013. View Article : Google Scholar : PubMed/NCBI
14	Cardeno A, Sánchez-Hidalgo M, Rosillo MA and Alarcón de la Lastra C: Oleuropein, a secoiridoid derived from olive tree, inhibits the proliferation of human colorectal cancer cell through downregulation of HIF-1α. Nutr Cancer. 65:147–156. 2013. View Article : Google Scholar
15	Cotter TG: Apoptosis and cancer: the genesis of a research field. Nat Rev Cancer. 9:501–507. 2009. View Article : Google Scholar : PubMed/NCBI
16	Johnstone RW, Ruefli AA and Lowe SW: Apoptosis: a link between cancer genetics and chemotherapy. Cell. 108:153–164. 2002. View Article : Google Scholar : PubMed/NCBI
17	Kumar A, Rajendran V, Sethumadhavan R and Purohit R: AKT kinase pathway: a leading target in cancer research. Scientific World Journal. 2013:7561342013. View Article : Google Scholar : PubMed/NCBI
18	Pavlidou A and Vlahos NF: Molecular alterations of PI3K/Akt/mTOR pathway: a therapeutic target in endometrial cancer. Scientific World Journal. 2014:7097362014. View Article : Google Scholar : PubMed/NCBI
19	Franke TF, Hornik CP, Segev L, Shostak GA and Sugimoto C: PI3K/Akt and apoptosis: size matters. Oncogene. 22:8983–8998. 2003. View Article : Google Scholar : PubMed/NCBI
20	Datta SR, Dudek H, Tao X, et al: Akt phosphorylation of BAD couples survival signals to the cell-intrinsic death machinery. Cell. 91:231–241. 1997. View Article : Google Scholar : PubMed/NCBI
21	Hahn-Windgassen A, Nogueira V, Chen CC, Skeen JE, Sonenberg N and Hay N: Akt activates the mammalian target of rapamycin by regulating cellular ATP level and AMPK activity. J Biol Chem. 280:32081–32089. 2005. View Article : Google Scholar : PubMed/NCBI
22	Rokutanda S, Fujita T, Kanatani N, et al: Akt regulates skeletal development through GSK3, mTOR, and FoxOs. Dev Biol. 328:78–93. 2009. View Article : Google Scholar : PubMed/NCBI
23	Adams JM and Cory S: Life-or-death decisions by the Bcl-2 protein family. Trends Biochem Sci. 26:61–66. 2001. View Article : Google Scholar : PubMed/NCBI
24	Nicholson KM and Anderson NG: The protein kinase B/Akt signalling pathway in human malignancy. Cell Signal. 14:381–395. 2002. View Article : Google Scholar : PubMed/NCBI
25	Vlahos CJ, Matter WF, Hui KY and Brown RF: A specific inhibitor of phosphatidylinositol 3-kinase, 2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one(LY294002). J Biol Chem. 269:5241–5248. 1994.PubMed/NCBI
26	Nogueira V, Park Y, Chen CC, et al: Akt determines replicative senescence and oxidative or oncogenic premature senescence and sensitizes cells to oxidative apoptosis. Cancer Cell. 14:458–470. 2008. View Article : Google Scholar : PubMed/NCBI
27	Robey RB and Hay N: Is Akt the ‘Warburg kinase’? - Akt-energy metabolism interactions and oncogenesis. Semin Cancer Biol. 19:25–31. 2009. View Article : Google Scholar : PubMed/NCBI
28	Fini L, Hotchkiss E, Fogliano V, et al: Chemopreventive properties of pinoresinol-rich olive oil involve a selective activation of the ATM-p53 cascade in colon cancer cell lines. Carcinogenesis. 29:139–146. 2008. View Article : Google Scholar
29	La Vecchia C: Association between Mediterranean dietary patterns and cancer risk. Nutr Rev. 67(Suppl 1): S126–S129. 2009. View Article : Google Scholar : PubMed/NCBI
30	Menendez JA, Papadimitropoulou A, Vellon L and Lupu R: A genomic explanation connecting ‘Mediterranean diet’, olive oil and cancer: oleic acid, the main monounsaturated fatty acid of olive oil, induces formation of inhibitory ‘PEA3 transcription factor-PEA3 DNA binding site’ complexes at the Her-2/neu (erbB-2) oncogene promoter in breast, ovarian and stomach cancer cells. Eur J Cancer. 42:2425–2432. 2006. View Article : Google Scholar : PubMed/NCBI
31	Kaufmann SH and Earnshaw WC: Induction of apoptosis by cancer chemotherapy. Exp Cell Res. 256:42–49. 2000. View Article : Google Scholar : PubMed/NCBI
32	Wiita AP, Ziv E, Wiita PJ, et al: Global cellular response to chemotherapy-induced apoptosis. Elife. 29:e012362013.
33	Czerski L and Nunez G: Apoptosome formation and caspase activation: is it different in the heart? J Mol Cell Cardiol. 37:643–652. 2004. View Article : Google Scholar : PubMed/NCBI
34	Fulda S and Debatin KM: Apoptosis signaling in tumor therapy. Ann NY Acad Sci. 1028:150–156. 2004. View Article : Google Scholar
35	Zhang L, Yu J, Park BH, Kinzler KW and Vogelstein B: Role of BAX in the apoptotic response to anticancer agents. Science. 290:989–992. 2000. View Article : Google Scholar : PubMed/NCBI
36	Fresno Vara JA, Casado E, de Castro J, Cejas P, Belda-Iniesta C and González-Barón M: PI3K/Akt signalling pathway and cancer. Cancer Treat Rev. 30:193–204. 2004. View Article : Google Scholar : PubMed/NCBI
37	Hennessy BT, Smith DL, Ram PT, Lu Y and Mills GB: Exploiting the PI3K/AKT pathway for cancer drug discovery. Nat Rev Drug Discov. 4:988–1004. 2005. View Article : Google Scholar : PubMed/NCBI
38	Minguez B, Tovar V, Chiang D, Villanueva A and Llovet JM: Pathogenesis of hepatocellular carcinoma and molecular therapies. Curr Opin Gastroenterol. 25:186–194. 2009. View Article : Google Scholar : PubMed/NCBI
39	Saxena NK, Sharma D, Ding X, et al: Concomitant activation of the JAK/STAT, PI3K/AKT, and ERK signaling is involved in leptin-mediated promotion of invasion and migration of hepatocellular carcinoma cells. Cancer Res. 67:2497–2507. 2007. View Article : Google Scholar : PubMed/NCBI
40	Wang GL, Iakova P, Wilde M, Awad S and Timchenko NA: Liver tumors escape negative control of proliferation via PI3K/Akt-mediated block of C/EBP alpha growth inhibitory activity. Genes Dev. 18:912–925. 2004. View Article : Google Scholar : PubMed/NCBI
41	Nogueira V, Park Y, Chen CC, et al: Akt determines replicative senescence and oxidative or oncogenic premature senescence and sensitizes cells to oxidative apoptosis. Cancer Cell. 14:458–470. 2008. View Article : Google Scholar : PubMed/NCBI
42	Liu J, Zhou J and Xing D: Phosphatidylinositol 3-kinase plays a vital role in regulation of rice seed vigor via altering NADPH oxidase activity. PLoS One. 7:e338172012. View Article : Google Scholar : PubMed/NCBI