The ERBB receptor inhibitor dacomitinib suppresses proliferation and invasion of pancreatic ductal adenocarcinoma cells
- 327 Downloads
Pancreatic ductal adenocarcinoma (PDAC), the most common malignancy of the pancreas, is the fourth most common cause of cancer-related death in the USA. Local progression, early tumor dissemination and low efficacy of current treatments are the major reasons for its high mortality rate. The ERBB family is over-expressed in PDAC and plays essential roles in its tumorigenesis; however, single-targeted ERBB inhibitors have shown limited activity in this disease. Here, we examined the anti-tumor activity of dacomitinib, a pan-ERBB receptor inhibitor, on PDAC cells.
Anti-proliferative effects of dacomitinib were determined using a cell proliferation assay and crystal violet staining. Annexin V/PI staining, radiation therapy and cell migration and invasion assays were carried out to examine the effects of dacomitinib on apoptosis, radio-sensitivity and cell motility, respectively. Quantitative reverse transcription-PCR (qRT-PCR) and Western blot analyses were applied to elucidate the molecular mechanisms underlying the anti-tumor activity of dacomitinib.
We found that dacomitinib diminished PDAC cell proliferation via inhibition of FOXM1 and its targets Aurora kinase B and cyclin B1. Moreover, we found that dacomitinib induced apoptosis and potentiated radio-sensitivity via inhibition of the anti-apoptotic proteins survivin and MCL1. Treatment with dacomitinib attenuated cell migration and invasion through inhibition of the epithelial-to-mesenchymal transition (EMT) markers ZEB1, Snail and N-cadherin. In contrast, we found that the anti-tumor activity of single-targeted ERBB agents including cetuximab (anti-EGFR mAb), trastuzumab (anti-HER2 mAb), H3.105.5 (anti-HER3 mAb) and erlotinib (EGFR small molecule inhibitor) were marginal.
Our findings indicate that dacomitinib-mediated blockade of the ERBB receptors yields advantages over single-targeted ERBB inhibition and provide a rationale for further investigation of the therapeutic potential of dacomitinib in the treatment of ERBB-driven PDAC.
KeywordsPancreatic ductal adenocarcinoma The ERBB family Dacomitinib Radio-sensitivity
This study was financially supported by a grant from the Hematology/Oncology and Stem Cell Transplantation Research Centre, Shariati Hospital, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran. Technical assistance of Ms. Azam Zaghal is gratefully acknowledged.
M.M. designed the study; F.E., H.Y., S.J., Z.A., S.H.M., D.B. and F.K. conducted the research; A.R.D., S.M.T., J.T., P.H., K.A. and A.G. analyzed the data; M.M., S.H. and V.V. wrote the paper; M.M. and S.H.G. were primarily responsible for the final content. All authors have reviewed and approved the final manuscript.
Compliance with ethical standards
The authors declare no conflict of interest.
- 7.D. Li and E.M. O’Reilly, in Seminars in oncology, (Elsevier, 2015), p. 134–143Google Scholar
- 9.M.J. Moore, D. Goldstein, J. Hamm, A. Figer, J.R. Hecht, S. Gallinger, H.J. Au, P. Murawa, D. Walde, R.A. Wolff, Erlotinib plus gemcitabine compared with gemcitabine alone in patients with advanced pancreatic cancer: A phase III trial of the National Cancer Institute of Canada clinical trials group. J. Clin. Oncol. 25, 1960–1966 (2007)CrossRefPubMedGoogle Scholar
- 10.H.Q. Xiong, A. Rosenberg, A. LoBuglio, W. Schmidt, R.A. Wolff, J. Deutsch, M. Needle, J.L. Abbruzzese, Cetuximab, a monoclonal antibody targeting the epidermal growth factor receptor, in combination with gemcitabine for advanced pancreatic cancer: A multicenter phase II trial. J. Clin. Oncol. 22, 2610–2616 (2004)CrossRefPubMedGoogle Scholar
- 11.J. Harder, G. Ihorst, V. Heinemann, R. Hofheinz, M. Moehler, P. Buechler, G. Kloeppel, C. Röcken, M. Bitzer, S. Boeck, Multicentre phase II trial of trastuzumab and capecitabine in patients with HER2 overexpressing metastatic pancreatic cancer. Br. J. Cancer 106, 1033–1038 (2012)CrossRefPubMedPubMedCentralGoogle Scholar
- 17.R.K. Schmidt-Ullrich, J.N. Contessa, G. Lammering, G. Amorino, P.-S. Lin, ERBB receptor tyrosine kinases and cellular radiation responses. Oncogene 22, 5855–5865 (2003)Google Scholar
- 18.N. Ioannou, A.M. Seddon, A. Dalgleish, D. Mackintosh, H. Modjtahedi, Expression pattern and targeting of HER family members and IGF-IR in pancreatic cancer. Front. Biosci. (Landmark Ed) 17, 2698–2724 (2012)Google Scholar
- 19.S. Ueda, S. Ogata, H. Tsuda, N. Kawarabayashi, M. Kimura, Y. Sugiura, S. Tamai, O. Matsubara, K. Hatsuse, H. Mochizuki, The correlation between cytoplasmic overexpression of epidermal growth factor receptor and tumor aggressiveness: Poor prognosis in patients with pancreatic ductal adenocarcinoma. Pancreas 29, e1–e8 (2004)Google Scholar
- 23.A.J. Saxby, A. Nielsen, C.J. Scarlett, A. Clarkson, A. Morey, A. Gill, R.C. Smith, Assessment of HER-2 status in pancreatic adenocarcinoma: Correlation of immunohistochemistry, quantitative real-time RT-PCR, and FISH with aneuploidy and survival. Am. J. Surg. Pathol. 29, 1125–1134 (2005)CrossRefPubMedGoogle Scholar
- 24.T. Hirakawa, B. Nakata, R. Amano, K. Kimura, S. Shimizu, G. Ohira, N. Yamada, M. Ohira, K. Hirakawa, HER3 overexpression as an independent indicator of poor prognosis for patients with curatively resected pancreatic cancer. Oncology 81, 192–198 (2011)Google Scholar
- 26.A.J. Gonzales, K.E. Hook, I.W. Althaus, P.A. Ellis, E. Trachet, A.M. Delaney, P.J. Harvey, T.A. Ellis, D.M. Amato, J.M. Nelson, Antitumor activity and pharmacokinetic properties of PF-00299804, a second-generation irreversible pan-erbB receptor tyrosine kinase inhibitor. Mol. Cancer Ther. 7, 1880–1889 (2008)CrossRefPubMedGoogle Scholar
- 27.Y.-L. Wu, Y. Cheng, X. Zhou, K.H. Lee, K. Nakagawa, S. Niho, F. Tsuji, R. Linke, R. Rosell, J. Corral, Dacomitinib versus gefitinib as first-line treatment for patients with EGFR-mutation-positive non-small-cell lung cancer (ARCHER 1050): A randomised, open-label, phase 3 trial. Lancet Oncol. 18, 1454–1466 (2017)CrossRefPubMedGoogle Scholar
- 28.T.S. Mok, Y. Cheng, X. Zhou, K.H. Lee, K. Nakagawa, S. Niho, M. Lee, R. Linke, R. Rosell, J. Corral, Improvement in overall survival in a randomized study that compared dacomitinib with gefitinib in patients with advanced non–small-cell lung cancer and EGFR-activating mutations. J. Clin. Oncol. 36, 2244–2250 (2018)CrossRefPubMedGoogle Scholar
- 29.J.A. Engelman, K. Zejnullahu, C.-M. Gale, E. Lifshits, A.J. Gonzales, T. Shimamura, F. Zhao, P.W. Vincent, G.N. Naumov, J.E. Bradner, PF00299804, an irreversible pan-ERBB inhibitor, is effective in lung cancer models with EGFR and ERBB2 mutations that are resistant to gefitinib. Cancer Res. 67, 11924–11932 (2007)CrossRefPubMedGoogle Scholar
- 30.H.-J. Nam, K.A. Ching, J. Kan, H.-P. Kim, S.-W. Han, S.-A. Im, T.-Y. Kim, J.G. Christensen, D.-Y. Oh, Y.-J. Bang, Evaluation of the antitumor effects and mechanisms of PF00299804, a pan-HER inhibitor, alone or in combination with chemotherapy or targeted agents in gastric cancer. Mol. Cancer Ther. 11, 439–451 (2012)CrossRefPubMedGoogle Scholar
- 32.F. Ather, H. Hamidi, M.S. Fejzo, S. Letrent, R.S. Finn, F. Kabbinavar, C. Head, S.G. Wong, Dacomitinib, an irreversible Pan-ErbB inhibitor significantly abrogates growth in head and neck cancer models that exhibit low response to cetuximab. PLoS One 8, e56112 (2013)CrossRefPubMedPubMedCentralGoogle Scholar
- 33.M. Momeny, G. Zarrinrad, F. Moghaddaskho, A. Poursheikhani, G. Sankanian, A. Zaghal, S. Mirshahvaladi, F. Esmaeili, H. Eyvani, F. Barghi, Dacomitinib, a pan-inhibitor of ErbB receptors, suppresses growth and invasive capacity of chemoresistant ovarian carcinoma cells. Sci. Rep. 7, 4204 (2017)CrossRefPubMedPubMedCentralGoogle Scholar
- 35.M. Momeny, J.M. Saunus, F. Marturana, A.E.M. Reed, D. Black, G. Sala, S. Iacobelli, J.D. Holland, D. Yu, L. Da Silva, Heregulin-HER3-HER2 signaling promotes matrix metalloproteinase-dependent blood-brain-barrier transendothelial migration of human breast cancer cell lines. Oncotarget 6, 3932 (2015)CrossRefPubMedPubMedCentralGoogle Scholar
- 38.S. Lu, F. Concha-Benavente, G. Shayan, R.M. Srivastava, S.P. Gibson, L. Wang, W.E. Gooding, R.L. Ferris, STING activation enhances cetuximab-mediated NK cell activation and DC maturation and correlates with HPV+ status in head and neck cancer. Oral Oncol. 78, 186–193 (2018)CrossRefPubMedPubMedCentralGoogle Scholar
- 43.J. Berthelet, L. Dubrez, Regulation of apoptosis by inhibitors of apoptosis (IAPs). Cells 2, 163–187 (2013)Google Scholar
- 47.N.M. Aiello, T. Brabletz, Y. Kang, M.A. Nieto, R.A. Weinberg, B.Z. Stanger, Upholding a role for EMT in pancreatic cancer metastasis. Nature 547, E7 (2017)Google Scholar
- 54.P.J. Loehrer Sr., Y. Feng, H. Cardenes, L. Wagner, J.M. Brell, D. Cella, P. Flynn, R.K. Ramanathan, C.H. Crane, S.R. Alberts, Gemcitabine alone versus gemcitabine plus radiotherapy in patients with locally advanced pancreatic cancer: An eastern cooperative oncology group trial. J. Clin. Oncol. 29, 4105–4112 (2011)CrossRefPubMedPubMedCentralGoogle Scholar
- 56.J. Maurel, M. Martin-Richard, C. Conill, M. Sánchez, L. Petriz, A. Ginès, R. Miquel, R. Gallego, R. Cajal, C. Ayuso, Phase I trial of gefitinib with concurrent radiotherapy and fixed 2-h gemcitabine infusion, in locally advanced pancreatic cancer. Int. J. Radiat. Oncol. Biol. Phys. 66, 1391–1398 (2006)CrossRefPubMedGoogle Scholar
- 57.L. Zhang, S. Yuan, Expression of c-erbB-2 oncogene protein, epidermal growth factor receptor, and TGF-beta1 in human pancreatic ductal adenocarcinoma. Hepatobiliary Pancreat. Dis. Int. 1, 620–623 (2002)Google Scholar
- 58.M.E. Valsecchi, M. McDonald, J.R. Brody, T. Hyslop, B. Freydin, C.J. Yeo, C. Solomides, S.C. Peiper, A.K. Witkiewicz, Epidermal growth factor receptor and insulinlike growth factor 1 receptor expression predict poor survival in pancreatic ductal adenocarcinoma. Cancer 118, 3484–3493 (2012)Google Scholar
- 64.K. Kimura, T. Sawada, M. Komatsu, M. Inoue, K. Muguruma, T. Nishihara, Y. Yamashita, N. Yamada, M. Ohira, K. Hirakawa, Antitumor effect of trastuzumab for pancreatic cancer with high HER-2 expression and enhancement of effect by combined therapy with gemcitabine. Clin. Cancer Res. 12, 4925–4932 (2006)CrossRefPubMedGoogle Scholar
- 66.H.J. Jacobsen, T.T. Poulsen, A. Dahlman, I. Kjær, K. Koefoed, J.W. Sen, D. Weilguny, B. Bjerregaard, C.R. Andersen, I.D. Horak, Pan-HER, an antibody mixture simultaneously targeting EGFR, HER2, and HER3, effectively overcomes tumor heterogeneity and plasticity. Clin. Cancer Res. 21, 4110–4122 (2015)CrossRefPubMedGoogle Scholar
- 68.M. Martin, F.A. Holmes, B. Ejlertsen, S. Delaloge, B. Moy, H. Iwata, G. von Minckwitz, S.K. Chia, J. Mansi, C.H. Barrios, Neratinib after trastuzumab-based adjuvant therapy in HER2-positive breast cancer (ExteNET): 5-years analysis of a randomised, double-blind, placebo-controlled, phase 3 trial. Lancet Oncol. 18, 1688–1700 (2017)CrossRefPubMedGoogle Scholar
- 69.K.L. Reckamp, G. Giaccone, D.R. Camidge, S.M. Gadgeel, F.R. Khuri, J.A. Engelman, M. Koczywas, A. Rajan, A.K. Campbell, D. Gernhardt, A phase 2 trial of dacomitinib (PF-00299804), an oral, irreversible pan-HER (human epidermal growth factor receptor) inhibitor, in patients with advanced non–small cell lung cancer after failure of prior chemotherapy and erlotinib. Cancer 120, 1145–1154 (2014)CrossRefPubMedPubMedCentralGoogle Scholar
- 72.A. Aghdassi, M. Sendler, A. Guenther, J. Mayerle, C.-O. Behn, C.-D. Heidecke, H. Friess, M. Büchler, M. Evert, M.M. Lerch, Recruitment of histone deacetylases HDAC1 and HDAC2 by the transcriptional repressor ZEB1 downregulates E-cadherin expression in pancreatic cancer. Gut 61, 439–448 (2012)CrossRefPubMedGoogle Scholar
- 78.M. Momeny, M. Malehmir, M. Zakidizaji, R. Ghasemi, H. Ghadimi, M.A. Shokrgozar, A.H. Emami, S. Nafissi, A. Ghavamzadeh, S.H. Ghaffari, Silibinin inhibits invasive properties of human glioblastoma U87MG cells through suppression of cathepsin B and nuclear factor kappa B-mediated induction of matrix metalloproteinase 9. Anti-Cancer Drugs 21, 252–260 (2010)CrossRefPubMedGoogle Scholar
- 79.D. Brix, K. Clemmensen, T. Kallunki, When good turns bad: Regulation of invasion and metastasis by ErbB2 receptor tyrosine kinase. Cells 3, 53–78 (2014)Google Scholar
- 83.H.-T. Guan, X.-H. Xue, Z.-J. Dai, X.-J. Wang, A. Li, Z.-Y. Qin, Down-regulation of survivin expression by small interfering RNA induces pancreatic cancer cell apoptosis and enhances its radiosensitivity. World J. Gastroenterol. 12, 2901–2907 (2006)Google Scholar
- 84.S.-H. Wei, K. Dong, F. Lin, X. Wang, B. Li, J.-j. Shen, Q. Zhang, R. Wang, H.-Z. Zhang, Inducing apoptosis and enhancing chemosensitivity to gemcitabine via RNA interference targeting mcl-1 gene in pancreatic carcinoma cell. Cancer Chemother. Pharmacol. 62, 1055–1064 (2008)CrossRefPubMedGoogle Scholar
- 87.P. Seshacharyulu, M.P. Ponnusamy, S. Rachagani, I. Lakshmanan, D. Haridas, Y. Yan, A.K. Ganti, S.K. Batra, Targeting EGF-receptor (s)-STAT1 axis attenuates tumor growth and metastasis through downregulation of MUC4 mucin in human pancreatic cancer. Oncotarget 6, 5164 (2015)CrossRefPubMedGoogle Scholar