The major vault protein mediates resistance to epidermal growth factor receptor inhibition in human hepatoma cells
Introduction
Liver cancer is the fifth most common cancer worldwide, with more than 500,000 new cases per year [1]. Complete surgical resection of tumors or liver transplantation is only possible in a minority of patients; for patients with advanced disease, the prognosis is extremely poor, with an overall median survival of only a few months. Response rates to classical chemotherapy are low, and even with combination regimens, durable remission has remained elusive [2]. Thus, there is a strong need for additional therapeutic options.
In recent years, rationally designed, molecularly targeted drugs have become available. These agents are designed to target growth or survival pathways hyper-activated in cancer cells. Tyrosine kinases are thought to be excellent molecular oncology targets because they transduce growth and survival signals and are hyper-activated in most, if not all, human malignancies [3]. The ErbB receptor tyrosine kinase family, comprising EGFR (=ErbB1) and ErbB2, -3, and -4, has been of central interest in the development of targeted anticancer strategies. Trastuzumab (Herceptin), a monoclonal antibody against ErbB2, is successfully being used in patients with ErbB2-overexpressing breast cancer, and overexpression of ErbB2 through gene amplification is a good predictor of favorable response [4]. Several preclinical and clinical studies have addressed the efficacy of EGFR-targeting agents, including tyrosine kinase inhibitors (TKIs), such as gefitinib (Iressa, ZD1839) and erlotinib (Tarceva, OSI-774), as well as monoclonal anti-EGFR antibodies, such as cetuximab, for the treatment of non-small cell lung cancer (NSCLC), head and neck cancer, colon carcinoma, glioblastoma, and other tumors [5], [6]. Although NSCLC patients with activating mutations in the kinase domain of EGFR respond favorably to EGFR TKIs, leading to their approval for this subset of malignancies, the molecular basis determining the response of tumor cells to EGFR-targeting drugs in other settings is only partially understood and is discussed controversially.
In HCC, the small molecule inhibitor sorafenib (Nexavar, BAY 43-9006), which targets multiple kinases, including Raf, c-kit, vascular endothelial growth factor receptor (VEGFR), and platelet-derived growth factor receptor (PDGFR), is the first agent that has been found to improve survival of patients with advanced disease [7], which demonstrates that targeted agents may improve the treatment opportunities for this devastating disease [8]. Several lines of evidence also argue for an important role of deregulated EGFR signaling in HCC development. EGFR and its ligands TGFα, HB-EGF, amphiregulin and betacellulin are elevated in HCC [9], [10], [11], and have been demonstrated to drive liver tumorigenesis in cell culture and animal models [12], [13], [14]. Hepatoma cells have been shown to respond with growth inhibition and cell death induction to inhibition of EGFR with gefitinib [15], [16], erlotinib [17] or cetuximab [18], and gefitinib prevented HCC development in cirrhotic rat livers [19]. Clinical phase II trials with EGFR inhibitors, however, have shown that cetuximab has no antitumor activity and that erlotinib improved disease control in only a subset of patients [20], [21], [22], raising the question as to which mechanisms determine the response of HCC to EGFR inhibition.
In the present study, we have used a panel of extensively characterized HCC-derived cell lines [23], [24] to identify molecular factors contributing to resistance against the EGFR inhibitor gefitinib. Our data demonstrate a previously unrecognized role of the major vault protein (MVP) for EGFR-inhibitor resistance in hepatoma cells.
Section snippets
Chemicals
Gefitinib was kindly provided by AstraZeneca (London, UK). All other chemicals were from Sigma (St. Louis, MO), unless stated otherwise.
Cell lines and cell culture
Hep3B human hepatocellular carcinoma cells were grown in RPMI-1640 medium (Invitrogen, Carlsbad, CA) with 10% fetal calf serum (FCS). HepG2 human hepatoblastoma cells were grown in minimal essential medium (MEM, Invitrogen) with 10% FCS plus 1 mM sodium pyruvate and 1% non-essential amino acids. The HCC1.2, HCC1.1, HCC2, and HCC3 cell lines were established and
Sensitivity to the EGFR inhibitor gefitinib widely differs between HCC cell lines
Six hepatoma cell lines, HCC2, HCC3, HCC1.2, HCC1.1, Hep3B and HepG2, were treated with increasing concentrations of gefitinib and subjected to MTT assays. HCC2, HCC3, HCC1.2 and HCC1.1 were established from HCC surgery specimens in our institute and have maintained many characteristics of the original tumors [23]. HCC1.1 and 1.2 were established from the same patient and were recently described as a human model of hepatocellular epithelial-to-mesenchymal transition (EMT) [28]. Gefitinib showed
Discussion
Conventional chemotherapy has been largely ineffective in HCC [2]. Recent success with the multikinase inhibitor sorafenib has demonstrated that targeted agents can improve treatment strategies for HCC. There is substantial evidence suggesting EGFR as a rational target for HCC therapy, as it has been reported to be associated with liver cancer development and recurrence [33]. Although single-agent treatment with the EGFR-targeting agent erlotinib has not been successful, combinations of
Acknowledgements
This work was supported by grants from the Jubilaeumsfonds der Österreichischen Nationalbank to M.G. (Projects 9595 and 14211) and from the Initiative for Cancer Research of the Medical University of Vienna to M.G., B.M., T.G. and W.B.
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Cited by (24)
CHD4 promotes acquired chemoresistance and tumor progression by activating the MEK/ERK axis
2023, Drug Resistance UpdatesNovel cancer stem cell marker MVP enhances temozolomide-resistance in glioblastoma
2022, Translational OncologyCitation Excerpt :With respect to malignant progression, increased levels of MVP and vault particles have been reported in several cancer types. However, it is still controversial that the increased MVP/vault expression is related to chemotherapy resistance [38], but it is certain that MVP is associated with resistance according to several papers [10,22]. MVP is a major component of the vault complex, which plays a pivotal role in chemoresistance by allowing intracellular drugs to enter the nucleus and by regulating MAPK/ERK and phosphoinositide 3-kinase/Akt signaling [17,23,24].
Wilms' tumor 1 enhances Cisplatin-resistance of advanced NSCLC
2014, FEBS LettersCitation Excerpt :Min Han et al. reported that down-regulation of MVP may allow the intracellular drug to enter the nucleus, which eliminated one of the best characterized barriers to chemotherapy [37]. Losert et al. demonstrated that in HCC, MVP was a key marker of sensitivity, suggesting MVP a novel player in resistance against EGFR inhibition [38]. Izquierdo et al. has clarified that MVP owned a greatest individual value as a marker of in vitro resistance to Cisplatin and Carboplatin [39,40].
Proteomic differences between hepatocellular carcinoma and nontumorous liver tissue investigated by a combined gel-based and label-free quantitative proteomics study
2013, Molecular and Cellular ProteomicsCitation Excerpt :It has been associated with several signaling pathways, including PI3K/Akt, MAPK, and STAT suggesting regulatory roles in these signaling processes (83–85). More recently, MVP has been found to be involved in resistance to epidermal growth factor inhibition of several HCC-derived cell lines (86). In our current study, we observed a significant up-regulation of MVP in HCC tissue that was verified in two independent patient cohorts.
Molecular characterization of EGFR and EGFRvIII signaling networks in human glioblastoma tumor xenografts
2012, Molecular and Cellular ProteomicsCitation Excerpt :MVP has been suggested to contribute to resistance toward chemotherapeutic agents in lung cancer, as the name suggests (44, 45). MVP expression is significantly increased across malignant brain tumors when compared with nonmalignant brain tissues (46), and MVP expression correlates with EGFR inhibitor (gefitinib) resistance (45), suggesting that increased expression of MVP may contribute to therapeutic resistance in EGFRvIII expressing tumors. GBP1 is a large GTP-binding interferon-inducible protein belonging to the dynamin family (47).
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These authors contributed equally to the study.