Reactive oxygen species‑mediated activation of the Src‑epidermal growth factor receptor‑Akt signaling cascade prevents bortezomib‑induced apoptosis in hepatocellular carcinoma cells

Hou,Jinlin; Cui,Anguo; Song,Peiying; Hua,Hui; Luo,Ting; Jiang,Yangfu

doi:10.3892/mmr.2014.2736

Reactive oxygen species‑mediated activation of the Src‑epidermal growth factor receptor‑Akt signaling cascade prevents bortezomib‑induced apoptosis in hepatocellular carcinoma cells

Authors:
- Jinlin Hou
- Anguo Cui
- Peiying Song
- Hui Hua
- Ting Luo
- Yangfu Jiang
View Affiliations

Affiliations: State Key Labaoratory of Biotherapy, Section of Oncogene, Chengdu, Sichuan 610041, P.R. China, Laboratory of Stem Cell Biology, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China, Cancer Center, West China Hospital, Sichuan University, Chengdu, Sichuan 610041, P.R. China
Published online on: October 22, 2014 https://doi.org/10.3892/mmr.2014.2736
Pages: 712-718

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

Proteasomes are essential for numerous cellular processes, including the cell cycle, regulation of gene expression and responses to cellular stress. Proteasome inhibitors are promising anticancer agents. The proteasome inhibitor bortezomib effectively suppresses certain types of cancer, including multiple myeloma and mantle cell lymphoma. However, bortezomib poorly inhibits solid tumors, including hepatocellular carcinoma. The activation of Akt represents an adaptive response that prevents bortezomib‑induced cell apoptosis. In the present study, bortezomib induced phosphorylation of EGFR, Src and Akt in hepatoma cells and inhibition of Src reduced bortezomib‑induced EGFR and Akt phosphorylation. Treatment of hepatoma cells with bortezomib led to an increase in the levels of intracellular reactive oxygen species (ROS). The ROS scavenger N‑acetyl‑L‑cysteine inhibits bortezomib‑induced ROS production and abrogates the phosphorylation of Src, epidermal growth factor receptor and Akt. The combination of bortezomib and saracatinib, a Src inhibitor, synergistically induced hepatoma cell apoptosis. The present study concluded that ROS mediated the activation of the Src‑EGFR‑Akt cascade by bortezomib. The combination of the Src inhibitor and bortezomib holds promise in the treatment of hepatocellular carcinoma.

View Figures

View References

1	Walz J, Erdmann A, Kania M, Typke D, Koster AJ and Baumeister W: 26S proteasome structure revealed by three-dimensional electron microscopy. J Struct Biol. 121:19–29. 1998. View Article : Google Scholar : PubMed/NCBI
2	Kim HM, Yu Y and Cheng Y: Structure characterization of the 26S proteasome. Biochim Biophys Acta. 1809:67–79. 2011. View Article : Google Scholar : PubMed/NCBI
3	Pickart CM and Eddins MJ: Ubiquitin: structures, functions, mechanisms. Biochim Biophys Acta. 1695:55–72. 2004. View Article : Google Scholar : PubMed/NCBI
4	Pereira SG and Oakley F: Nuclear factor-κB1: regulation and function. Int J Biochem Cell Bio. 40:1425–1430. 2008.
5	Zhang M, Pickart CM and Coffino P: Determinants of proteasome recognition of ornithine decarboxylase, a ubiquitin-independent substrate. EMBO J. 22:1488–1496. 2003. View Article : Google Scholar : PubMed/NCBI
6	Tsvetkov P, Reuven N and Shaul Y: Ubiquitin-independent p53 proteasomal degradation. Cell Death Differ. 17:103–108. 2010. View Article : Google Scholar : PubMed/NCBI
7	Lee YS, Wan J, Kim BJ, Bae MA and Song BJ: Ubiquitin-dependent degradation of p53 protein despite phosphorylation at its N terminus by acetaminophen. J Pharmacol Exp Ther. 317:202–208. 2006. View Article : Google Scholar : PubMed/NCBI
8	Jariel-Encontre I, Bossis G and Piechaczyk M: Ubiquitin-independent degradation of proteins by the proteasome. Biochim Biophys Acta. 1786:153–177. 2008.PubMed/NCBI
9	Papandreou CN and Logothetis CJ: Bortezomib as a potential treatment for prostate cancer. Cancer Res. 64:5036–5043. 2004. View Article : Google Scholar : PubMed/NCBI
10	Richardson PG and Anderson KC: Bortezomib: a novel therapy approved for multiple myeloma. Clin Adv Hematol Oncol. 1:596–600. 2003.PubMed/NCBI
11	O’Connor OA: Marked clinical activity of the proteasome inhibitor bortezomib in patients with follicular and mantle-cell lymphoma. Clin Lymphoma Myeloma. 6:191–199. 2005.
12	Ko BS, Chang TC, Chen CH, et al: Bortezomib suppresses focal adhesion kinase expression via interrupting nuclear factor-kappa B. Life Sci. 86:199–206. 2010. View Article : Google Scholar : PubMed/NCBI
13	Lara PN Jr, Davies AM, Mack PC, et al: Proteasome inhibition with PS-341 (bortezomib) in lung cancer therapy. Semin Oncol. 31:40–46. 2004. View Article : Google Scholar : PubMed/NCBI
14	Chen KF, Yeh PY, Yeh KH, Lu YS, Huang SY and Cheng AL: Downregulation of phospho-Akt is a major molecular determinant of bortezomib-induced apoptosis in hepatocellular carcinoma cells. Cancer Res. 68:6698–6707. 2008. View Article : Google Scholar : PubMed/NCBI
15	Lin YC, Chen KC, Chen CC, Cheng AL and Chen KF: CIP2A-mediated Akt activation plays a role in bortezomib-induced apoptosis in head and neck squamous cell carcinoma cells. Oral Oncol. 48:585–593. 2012. View Article : Google Scholar : PubMed/NCBI
16	McCubrey JA, Steelman LS, Abrams SL, et al: Roles of the RAF/MEK/ERK and PI3K/PTEN/AKT pathways in malignant transformation and drug resistance. Adv Enzyme Regul. 46:249–279. 2006. View Article : Google Scholar : PubMed/NCBI
17	Haynes MP, Li L, Sinha D, et al: Src kinase mediates phosphatidylinositol 3-kinase/Akt-dependent rapid endothelial nitric-oxide synthase activation by estrogen. J Biol Chem. 278:2118–2123. 2003. View Article : Google Scholar : PubMed/NCBI
18	Mao W, Irby R, Coppola D, et al: Activation of c-Src by receptor tyrosine kinases in human colon cancer cells with high metastatic potential. Oncogene. 15:3083–3090. 1997. View Article : Google Scholar : PubMed/NCBI
19	Yang F, Jove V, Chang S, et al: Bortezomib induces apoptosis and growth suppression in human medulloblastoma cells, associated with inhibition of AKT and NF-κB signaling and synergizes with an ERK inhibitor. Cancer Biol Ther. 13:349–357. 2012.PubMed/NCBI
20	Markovic SN, Geyer SM, Dawkins F, et al: A phase II study of bortezomib in the treatment of metastatic malignant melanoma. Cancer. 103:2584–2589. 2005. View Article : Google Scholar : PubMed/NCBI
21	Vivanco I and Sawyers CL: The phosphatidylinositol 3-kinase AKT pathway in human cancer. Nat Rev Cancer. 2:489–501. 2002. View Article : Google Scholar : PubMed/NCBI
22	Hay N: The Akt-mTOR tango and its relevance to cancer. Cancer Cell. 8:179–183. 2005. View Article : Google Scholar : PubMed/NCBI
23	Junttila MR, Puustinen P, Niemelä M, et al: CIP2A inhibits PP2A in human malignancies. Cell. 130:51–62. 2007. View Article : Google Scholar : PubMed/NCBI
24	Chen K, Liu C, Lin Y, et al: CIP2A mediates effects of bortezomib on phospho-Akt and apoptosis in hepatocellular carcinoma cells. Oncogene. 29:6257–6266. 2010. View Article : Google Scholar : PubMed/NCBI
25	An J and Rettig MB: Epidermal growth factor receptor inhibition sensitizes renal cell carcinoma cells to the cytotoxic effects of bortezomib. Mol Cancer Ther. 6:61–69. 2007. View Article : Google Scholar : PubMed/NCBI
26	Lorch JH, Thomas TO and Schmoll HJ: Bortezomib inhibits cell-cell adhesion and cell migration and enhances epidermal growth factor receptor inhibitor-induced cell death in squamous cell cancer. Cancer Res. 67:727–734. 2007. View Article : Google Scholar : PubMed/NCBI
27	Codony-Servat J, Tapia MA, Bosch M, et al: Differential cellular and molecular effects of bortezomib, a proteasome inhibitor, in human breast cancer cells. Mol Cancer Ther. 5:665–675. 2006. View Article : Google Scholar : PubMed/NCBI
28	Sloss CM, Wang F, Liu R, et al: Proteasome inhibition activates epidermal growth factor receptor (EGFR) and EGFR-independent mitogenic kinase signaling pathways in pancreatic cancer cells. Clin Cancer Res. 14:5116–5123. 2008. View Article : Google Scholar
29	Cascone T, Morelli MP, Morgillo F, et al: Synergistic anti-proliferative and pro-apoptotic activity of combined therapy with bortezomib, a proteasome inhibitor, with anti-epidermal growth factor receptor (EGFR) drugs in human cancer cells. J Cell Physiol. 216:698–707. 2008. View Article : Google Scholar
30	Wilhelm SM, Adnane L, Newell P, Villanueva A, Llovet JM and Lynch M: Preclinical overview of sorafenib, a multikinase inhibitor that targets both Raf and VEGF and PDGF receptor tyrosine kinase signaling. Mol Cancer Ther. 7:3129–3140. 2008. View Article : Google Scholar : PubMed/NCBI
31	Chen KF, Yu HC, Liu TH, Lee SS, Chen PJ and Cheng AL: Synergistic interactions between sorafenib and bortezomib in hepatocellular carcinoma involve PP2A-dependent Akt inactivation. J Hepatol. 52:88–95. 2010. View Article : Google Scholar : PubMed/NCBI
32	Saad F and Lipton A: SRC kinase inhibition: targeting bone metastases and tumor growth in prostate and breast cancer. Cancer Treat Rev. 36:177–184. 2010. View Article : Google Scholar : PubMed/NCBI
33	Martinez Flores K, Uribe Marin BC, Souza Arroyo V, et al: Hepatocytes display a compensatory survival response against cadmium toxicity by a mechanism mediated by EGFR and Src. Toxicol In Vitro. 27:1031–1042. 2013.PubMed/NCBI
34	Yu C, Rahmani M, Dent P and Grant S: The hierarchical relationship between MAPK signaling and ROS generation in human leukemia cells undergoing apoptosis in response to the proteasome inhibitor Bortezomib. Exp Cell Res. 295:555–566. 2004. View Article : Google Scholar
35	Vasant C, Rajaram R and Ramasami T: Apoptosis of lymphocytes induced by chromium (VI/V) is through ROS-mediated activation of Src-family kinases and caspase-3. Free Radic Biol. 35:1082–1100. 2003. View Article : Google Scholar : PubMed/NCBI
36	Morgillo F, D’Aiuto E, Troiani T, et al: Antitumor activity of bortezomib in human cancer cells with acquired resistance to anti-epidermal growth factor receptor tyrosine kinase inhibitors. Lung Cancer. 71:283–290. 2011. View Article : Google Scholar
37	Kim A, Park S, Lee JE, et al: The dual PI3K and mTOR inhibitor NVP-BEZ235 exhibits anti-proliferative activity and overcomes bortezomib resistance in mantle cell lymphoma cells. Leuk Res. 36:912–920. 2012. View Article : Google Scholar : PubMed/NCBI