Top

Published in:

01-10-2013 | Original Article

Reversibility of regorafenib effects in hepatocellular carcinoma cells

Authors: Rosalba D’Alessandro, Maria G. Refolo, Catia Lippolis, Caterina Messa, Aldo Cavallini, Roberta Rossi, Leonardo Resta, Antonio Di Carlo, Brian I. Carr

Published in: Cancer Chemotherapy and Pharmacology | Issue 4/2013

Abstract

Purpose

Multikinase growth inhibitors inhibit their target kinases with varying potency. Patients often require lower doses or therapy breaks due to drug toxicities. To evaluate the effects of drug withdrawal on hepatocellular carcinoma cells after incubation with growth-inhibitory concentrations of regorafenib, cell growth, migration and invasion, and signaling were examined.

Methods

Cell proliferation, motility, and invasion were analyzed by MTT, wound healing, and invasion assays, respectively, and MAPK pathway protein markers were analyzed by Western blot.

Results

After regorafenib removal, cell growth, migration, and invasion recovered. Repeated drug exposure resulted in changes in cell growth patterns. Recovery could be blocked by sub-growth-inhibitory concentrations of either doxorubicin or vitamin K1. Recovery of growth was associated with increased phospho-JNK, phospho-p38, and phospho-STAT3 levels. The recovery of growth, migration, and signaling were blocked by a JNK inhibitor.

Conclusions

Removal of regorafenib from growth-inhibited cells resulted in a JNK-dependent recovery of growth and migration.

Available only for authorised users

Ahmad T, Eisen T (2004) Kinase inhibition with BAY 43-9006 in renal cell carcinoma. Clin Cancer Res 10:6388S–6392SPubMedCrossRef

Cervello M, Bachvarov D, Lampiasi N, Cusimano A, Azzolina A, McCubrey JA, Montalto G (2012) Molecular mechanisms of sorafenib action in liver cancer cells. Cell Cycle 11:2843–2855PubMedCrossRef

Wilhelm SM, Carter C, Tang L, Wilkie D, McNabola A, Rong H, Chen C, Zhang X, Vincent P, McHugh M, Cao Y, Shujath J, Gawlak S, Eveleigh D, Rowley B, Liu L, Adnane L, Lynch M, Auclair D, Taylor I, Gedrich R, Voznesensky A, Riedl B, Post LE, Bollag G, Trail PA (2004) BAY 43-9006 exhibits broad spectrum oral antitumor activity and targets the RAF/MEK/ERK pathway and receptor tyrosine kinases involved in tumor progression and angiogenesis. Cancer Res 64:7099–7109PubMedCrossRef

Liu L, Cao Y, Chen C, Zhang X, McNabola A, Wilkie D, Wilhelm S, Lynch M, Carter C (2006) Sorafenib blocks the RAF/MEK/ERK pathway, inhibits tumor angiogenesis, and induces tumor cell apoptosis in hepatocellular carcinoma model PLC/PRF/5. Cancer Res 66:11851–11858PubMedCrossRef

Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, de Oliveira AC, Santoro A, Raoul JL, Forner A, Schwartz M, Porta C, Zeuzem S, Bolondi L, Greten TF, Galle PR, Seitz JF, Borbath I, Häussinger D, Giannaris T, Shan M, Moscovici M, Voliotis D, Bruix J, SHARP Investigators Study Group (2008) Sorafenib in advanced hepatocellular carcinoma. N Engl J Med 359:378–390PubMedCrossRef

Cheng AL, Kang YK, Chen Z, Tsao CJ, Qin S, Kim JS, Luo R, Feng J, Ye S, Yang TS, Xu J, Sun Y, Liang H, Liu J, Wang J, Tak WY, Pan H, Burock K, Zou J, Voliotis D, Guan Z (2009) Efficacy and safety of sorafenib in patients in the Asia-Pacific region with advanced hepatocellular carcinoma: a phase III randomised, double-blind, placebo-controlled trial. Lancet Oncol 10:25–34PubMedCrossRef

Otsuka T, Eguchi Y, Kawazoe S, Yanagita K, Ario K, Kitahara K, Kawasoe H, Kato H, Mizuta T, The saga liver cancer study group (2012) Skin toxicities and survival in advanced hepatocellular carcinoma patients treated with sorafenib. Hepatol Res 42:879–886PubMedCrossRef

Edmonds K, Hull D, Spencer-Shaw A, Koldenhof J, Chrysou M, Boers-Doets C, Molassiotis A (2012) Strategies for assessing and managing the adverse events of sorafenib and other targeted therapies in the treatment of renal cell and hepatocellular carcinoma: recommendations from a European nursing task group. Eur J Oncol Nurs 16:172–184PubMedCrossRef

Porta C, Paglino C, Imarisio I, Bonomi L (2007) Uncovering Pandora’s vase: the growing problem of new toxicities from novel anticancer agents. The case of sorafenib and sunitinib. Clin Exp Med 7:127–134PubMedCrossRef

10.

Wilhelm SM, Dumas J, Adnane L, Lynch M, Carter CA, Schütz G, Thierauch KH, Zopf D (2011) Regorafenib (BAY 73-4506): a new oral multikinase inhibitor of angiogenic, stromal and oncogenic receptor tyrosine kinases with potent preclinical antitumor activity. Int J Cancer 129:245–255PubMedCrossRef

11.

Strumberg D, Schultheis B (2012) Regorafenib and cancer. Expert Opinion Invest Drugs 21:879–889CrossRef

12.

Demetri GD, Reichardt P, Kang YK, Blay JY, Rutkowski P, Gelderblom H, Hohenberger P, Leahy M, von Mehren M, Joensuu H, Badalamenti G, Blackstein M, Le Cesne A, Schöffski P, Maki RG, Bauer S, Nguyen BB, Xu J, Nishida T, Chung J, Kappeler C, Kuss I, Laurent D, Casali PG, GRID study investigators (2013) Efficacy and safety of regorafenib for advanced gastrointestinal stromal tumours after failure of imatinib and sunitinib (GRID): an international, multicentre, randomised, placebo-controlled, phase 3 trial. Lancet 381:295–302PubMedCrossRef

13.

Grothey A, Cutsem EV, Sobrero A, Siena S, Falcone A, Ychou M, Humblet Y, Bouché O, Mineur L, Barone C, Adenis A, Tabernero J, Yoshino T, Lenz HJ, Goldberg RM, Sargent DJ, Cihon F, Cupit L, Wagner A, Laurent D, CORRECT Study Group (2013) Regorafenib monotherapy for previously treated metastatic colorectal cancer (CORRECT): an international, multicentre, randomised, placebo-controlled, phase 3 trial. Lancet 381:303–312PubMedCrossRef

14.

Brahimi F, Rachid Z, Qiu Q, McNamee JP, Li YJ, Tari AM, Jean-Claude BJ (2004) Multiple mechanisms of action of ZR2002 in human breast cancer cells: a novel combi-molecule designed to block signaling mediated by the ERB family of oncogenes and to damage genomic DNA. Int J Cancer 112:484–491PubMedCrossRef

15.

Mancuso MR, Davis R, Norberg SM, O’Brien S, Sennino B, Nakahara T, Yao VJ, Inai T, Brooks P, Freimark B, Shalinsky DR, Hu-Lowe DD, McDonald DM (2006) Rapid vascular regrowth in tumors after reversal of VEGF inhibition. J Clin Invest 116:2610–2621PubMedCrossRef

16.

Steeghs N, Rabelink TJ, Op’t Roodt J, Batman E, Cluitmans FH, Weijl NI, de Koning E, Gelderblom H (2010) Reversibility of capillary density after discontinuation of bevacizumab treatment. Ann Oncol 21:1100–1105PubMedCrossRef

17.

Carr BI, Cavallini A, Lippolis C, D’Alessandro R, Messa C, Refolo MG, Tafaro A (2013) Fluoro-Sorafenib (Regorafenib) effects on hepatoma cells: growth inhibition, quiescence and recovery. J Cell Physiol 228:292–297PubMedCrossRef

18.

Sharma SV, Lee DY, Li B, Quinlan MP, Takahashi F, Maheswaran S, McDermott U, Azizian N, Zou L, Fischbach MA, Wong KK, Brandstetter K, Wittner B, Ramaswamy S, Classon M, Settleman J (2010) A chromatin-mediated reversible drug tolerant state in cancer cell subpopulations. Cell 141:69–80PubMedCrossRef

19.

Carr BI, D’Alessandro R, Refolo MG, Iacovazzi PA, Lippolis C, Messa C, Cavallini A, Correale M, Di Carlo A (2013) Effects of low concentrations of Regorafenib and Sorafenib on human HCC cell AFP, migration, invasion and growth in vitro. J Cell Physiol 228:1344–1350PubMedCrossRef

20.

Ebos JM, Lee CR, Cruz-Munoz W, Bjarnason GA, Christensen JG, Kerbel RS (2009) Accelerated metastasis after short-term treatment with a potent inhibitor of tumor angiogenesis. Cancer Cell 15:232–239PubMedCrossRef

21.

Cao H, Phan H, Yang LX (2012) Improved chemotherapy for hepatocellular carcinoma. Anticancer Res 32:1379–1386PubMed

22.

Varela M, Real MI, Burrel M, Forner A, Sala M, Brunet M, Ayuso C, Castells L, Montaná X, Llovet JM, Bruix J (2007) Chemoembolization of hepatocellular carcinoma with drug eluting beads: efficacy and doxorubicin pharmacokinetics. J Hepatology 46:474–481CrossRef

23.

Wei G, Wang M, Hyslop T, Wang Z, Carr BI (2010) Vitamin K enhancement of Sorafenib-mediated HCC cell growth inhibition in vitro and in vivo. Int J Cancer 127:2949–2958PubMedCrossRef

24.

Paraiso KH, Fedorenko IV, Cantini LP, Munko AC, Hall M, Sondak VK, Messina JL, Flaherty KT, Smalley KS (2010) Recovery of phospho-ERK activity allows melanoma cells to escape from BRAF inhibitor therapy. Br J Cancer 102:1724–1730PubMedCrossRef

25.

Gedaly R, Angulo P, Chen C, Creasy KT, Spear BT, Hundley J, Daily MF, Shah M, Evers BM (2012) The role of PI3 K/mTOR inhibition in combination with sorafenib in hepatocellular carcinoma treatment. Anticancer Res 32:2531–2536PubMed

26.

Ibrahim N, Yu Y, Walsh WR, Yang JL (2012) Molecular targeted therapies for cancer: sorafenib mono-therapy and its combination with other therapies. Oncol Rep 27:1303–1311PubMed

27.

Kudo M (2012) Targeted therapy for liver cancer: updated review in 2012. Curr Cancer Drug Targets 12:1062–1072PubMed

28.

Garlick DS, Greiner DL, Davis RJ (2011) The role of JNK in the development of hepatocellular carcinoma. Genes Dev 25:634–636PubMedCrossRef

29.

Bode AM, Dong Z (2007) The functional contrariety of JNK. Mol Carcinog 46:591–598PubMedCrossRef

30.

Lee YC, Chang AY, Lin-Feng MH, Tsou WI, Chiang IH, Lai MZ (2012) Paxillin phosphorylation by JNK and p38 is required for NFAT activation. Eur J Immunol 42:2165–2175PubMedCrossRef

31.

Zhang YH, Wang SQ, Sun CR, Wang M, Wang B, Tang JW (2011) Inhibition of JNK1 expression decreases migration and invasion of mouse hepatocellular carcinoma cell line in vitro. Med Oncol 28:966–972PubMedCrossRef

32.

Ranganathan AC, Zhang L, Adam AP, Aguirre-Ghiso JA (2006) Functional coupling of p38-dependent protein kinase-like endoplasmatic reticulum kinase to drug resistance of dormant carcinoma cells. Cancer Res 66:1702–1710PubMedCrossRef

33.

Wagner EF, Nebreda AR (2009) Signal integration by JNK and p38 MAPK pathway in cancer development. Nature Rev Cancer 9:537–549CrossRef

34.

Kim JH, Lee SC, Ro J, Kang HS, Kim HS, Yoon S (2010) Jnk signaling pathway-mediated regulation of Stat3 activation is linked to the development of doxorubicin resistance in cancer cell lines. Biochem Pharmacol 79:373–380PubMedCrossRef

Title: Reversibility of regorafenib effects in hepatocellular carcinoma cells
Authors: Rosalba D’Alessandro
Maria G. Refolo
Catia Lippolis
Caterina Messa
Aldo Cavallini
Roberta Rossi
Leonardo Resta
Antonio Di Carlo
Brian I. Carr
Publication date: 01-10-2013
Publisher: Springer Berlin Heidelberg
Published in: Cancer Chemotherapy and Pharmacology / Issue 4/2013
Print ISSN: 0344-5704
Electronic ISSN: 1432-0843
DOI: https://doi.org/10.1007/s00280-013-2269-8

Webinar | 19-02-2024 | 17:30 (CET)

Keynote webinar | Spotlight on antibody–drug conjugates in cancer

Watch now

Antibody–drug conjugates (ADCs) are novel agents that have shown promise across multiple tumor types. Explore the current landscape of ADCs in breast and lung cancer with our experts, and gain insights into the mechanism of action, key clinical trials data, existing challenges, and future directions.

Dr. Véronique Diéras

Prof. Fabrice Barlesi

Developed by: Springer Medicine

At a glance: The STEP trials

Springer Medicine

Abstract

Purpose

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 4/2013

Disruption of ZO-1/claudin-4 interaction in relation to inflammatory responses in methotrexate-induced intestinal mucositis

Inhibition of sorcin reverses multidrug resistance of K562/A02 cells and MCF-7/A02 cells via regulating apoptosis-related proteins

A multicenter phase II study of belotecan, a new camptothecin analogue, in elderly patients with previously untreated, extensive-stage small cell lung cancer

A clinical investigation of inhibitory effect of panobinostat on CYP2D6 substrate in patients with advanced cancer

Pharmacogenetic determinants associated with sunitinib-induced toxicity and ethnic difference in Korean metastatic renal cell carcinoma patients

Receptor tyrosine kinase alterations and therapeutic opportunities in squamous cell carcinoma of the lung

Keynote webinar | Spotlight on antibody–drug conjugates in cancer