Strong enhancement by IGF1-R antagonists of hepatocellular carcinoma cell migration inhibition by Sorafenib and/or vitamin K1
- 186 Downloads
Emerging evidence indicates that combining Sorafenib with vitamin K1 (VK1) may result in a synergistic inhibition of hepatocellular carcinoma (HCC) cell migration and proliferation. Despite this synergy, its benefits may be limited due to drug resistance resulting from cross-talk with the tumor microenvironment. Insulin-like growth factor-1 (IGF1) signaling acts as an important modulator of HCC cell growth, motility and drug resistance. Therefore, we aimed to explore the effects of Sorafenib in combination with VK1 and/or IGF1-R antagonists on HCC cells.
Scratch wound migration assays were performed to assess the motility of HCC-derived PLC/PRF/5, HLF and Hep3B cells. The synergistic, additive or antagonistic effects of Sorafenib, VK1 and IGF1-R antagonists on HCC cell motility were assessed using CompuSyn software. The effects mediated by these various compounds on HCC cytoskeleton organization were evaluated using DyLight 554 Phalloidin staining. Proliferation and migration-associated signaling pathways were analyzed in PLC/PRF/5 cells using Erk1/2 and Akt activation kits and Western blotting (Mek, JNK, Akt, Paxillin and p38), respectively.
The effects of the IGF1-R antagonists GSK1838705A and OSI-906 on HCC cell migration inhibition after Sorafenib and/or VK1 administration, individually or in combination, were evaluated. We found a synergistic effect in PLC/PRF/5, HLF and Hep3B cells for combinations of fixed doses of GSK1838705A or OSI-906 together with different doses of Sorafenib and/or VK1. The levels of synergy were found to be stronger at higher Sorafenib and/or VK1 concentrations and lower or absent at lower concentrations, with some variation among the different cell lines tested. In addition, we found that in PLC/PRF/5 and HLF cells IGF1-R blockage strongly enhanced the reduction and redistribution of F-actin induced by Sorafenib and/or VK1 through alterations in the phosphorylation levels of some of the principal proteins involved in the MAPK signaling cascade, which is essential for cell migration.
Our results indicate that modulation of the efficacy of Sorafenib through combinations with VK1 and/or IGF1-R antagonists results in synergistic inhibition of HCC cell migration.
KeywordsHepatocellular carcinoma Combination therapy Sorafenib Vitamin K1 IGF1 Microenvironment Cytoskeleton
This research was supported by the Italian Ministry of Public Health (n.11/2016).
This research was supported by the Italian Ministry of Public Health (n.11/2016).
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest.
- 2.M. Shimada, K. Takenaka, T. Gion, Y. Fujiwara, K. Kajiyama, T. Maeda, K. Shirabe, T. Nishizaki, K. Yanaga, K. Sugimachi, Prognosis of recurrent hepatocellular carcinoma: A 10-year surgical experience in Japan. Gastroenterology 111, 720-726 (1996). https://doi.org/10.1053/gast.1996.v111.pm8780578 CrossRefPubMedGoogle Scholar
- 4.S. Aishima, Y. Basaki, Y. Oda, Y. Kuroda, Y. Nishihara, K. Taguchi, A. Taketomi, Y. Maehara, F. Hosoi, Y. Maruyama, A. Fotovati, S. Oie, M. Ono, T. Ueno, M. Sata, H. Yano, M. Kojiro, M. Kuwano, M. Tsuneyoshi, High expression of insulin-like growth factor binding protein-3 is correlated with lower portal invasion and better prognosis in human hepatocellular carcinoma. Cancer Sci. 97, 1182-1190 (2006). https://doi.org/10.1111/j.1349-7006.2006.00322.x CrossRefPubMedGoogle Scholar
- 5.G. Giannelli, F. Pierri, P. Trerotoli, F. Marinosci, G. Serio, O. Schiraldi, S. Antonaci, Occurrence of portal vein tumor thrombus in epatocellular carcinoma affects prognosis and survival. A retrospettive clinical study of 150 cases. Hepatol. Res. 24, 50-59 (2002). https://doi.org/10.1016/S1386-6346(02)00027-X CrossRefPubMedGoogle Scholar
- 10.S.M. Wilhelm, C. Carter, L. Tang, D. Wilkie, A. McNabola, H. Rong, C. Chen, X. Zhang, P. Vincent, M. McHugh, Y. Cao, J. Shujath, S. Gawlak, D. Eveleigh, B. Rowley, L. Liu, L. Adnane, M. Lynch, D. Auclair, I. Taylor, R. Gedrich, A. Voznesensky, B. Riedl, L.E. Post, G. Bollag, P.A. Trail, 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, 7099-7109 (2004). https://doi.org/10.1158/0008-5472.CAN-04-1443 CrossRefPubMedGoogle Scholar
- 11.J.M. Llovet, S. Ricci, V. Mazzaferro, P. Hilgard, E. Gane, J.F. Blanc, A.C. de Oliveira, A. Santoro, J.L. Raoul, A. Forner, M. Schwartz, C. Porta, S. Zeuzem, L. Bolondi, T.F. Greten, P.R. Galle, J.F. Seitz, I. Borbath, D. Häussinger, T. Giannaris, M. Shan, M. Moscovici, D. Voliotis, J. Bruix, SHARP Investigators Study Group. Sorafenib in advanced hepatocellular carcinoma. N. Engl. J. Med. 359, 378–390 (2008)CrossRefPubMedGoogle Scholar
- 12.S.M. Wilhelm, L. Adnane, P. Newell, A. Villanueva, J.M. Llovet, M. Lynch, 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). https://doi.org/10.1158/1535-7163.MCT-08-0013 CrossRefPubMedGoogle Scholar
- 13.L. Gao, C. Shay, F. Lv, X. Wang, Y. Teng. Implications of FGF19 on sorafenib-mediated nitric oxide production in hepatocellular carcinoma cells - a short report. Cell. Oncol. 41, 85-91 (2018). https://doi.org/10.1007/s13402-017-0354-4.
- 19.T. Shibayama-Imazu, S. Sakairi, A. Watanabe, T. Aiuchi, S. Nakajo, K. Nakaya, Vitamin K(2) selectively induced apoptosis in ovarian TYK-nu and pancreatic MIA PaCa-2 cells out of eight solid tumor cell lines through a mechanism different from geranylgeraniol. J. Cancer Res. Clin. Oncol. 129, 1-11 (2003). https://doi.org/10.1007/s00432-002-0393-7 CrossRefPubMedGoogle Scholar
- 20.T. Yokoyama, K. Miyazawa, M. Naito, J. Toyotake, T. Tauchi, M. Itoh, A. You, Y. Hayashi, M.M. Georgescu, Y. Kondo, S. Kondo, K. Ohyashiki, Vitamin K2 induces autophagy and apoptosis simultaneously in leukemia cells. Autophagy 4, 629-640 (2008). https://doi.org/10.4161/auto.5941 CrossRefPubMedGoogle Scholar
- 22.S. Kuriyama, M. Hitomi, H. Yoshiji, T. Nonomura, T. Tsujimoto, A. Mitoro, T. Akahane, M. Ogawa, S. Nakai, T. Masaki, N. Uchida, Vitamins K2, K3 and K5 exert in vivo antitumor effects on hepatocellular carcinoma by regulating the expression of G1 phase-related cell cycle molecules. Int. J. Oncol. 27, 505-511 (2005)PubMedGoogle Scholar
- 26.R. D'Alessandro, M.G. Refolo, C. Lippolis, G. Giannuzzi, N. Carella, C. Messa, A. Cavallini, B.I. Carr, Antagonism of sorafenib and regorafenib actions by platelet factors in hepatocellular carcinoma cell lines. BMC Cancer 14, 351 (2014). https://doi.org/10.1186/1471-2407-14-351 CrossRefPubMedPubMedCentralGoogle Scholar
- 28.C. Lippolis, M.G. Refolo, R. D'Alessandro, N. Carella, C.A. Messa, A. Cavallini, B.I. Carr, Resistance to multikinase inhibitor actions mediated by insulin like growth factor-1. J. Exp. Clin. Cancer Res. 34, 90 (2015). https://doi.org/10.1186/s13046-015-0210-1 CrossRefPubMedPubMedCentralGoogle Scholar
- 29.P. Sabbatini, S. Korenchuk, J.L. Rowand, A. Groy, Q. Liu, D. Leperi, C. Atkins, M. Dumble, J. Yang, K. Anderson, R.G. Kruger, R.R. Gontarek, K.R. Maksimchuk, S. Suravajjala, R.R. Lapierre, J.B. Shotwell, J.W. Wilson, S.D. Chamberlain, S.K. Rabindran, R. Kumar, GSK1838705A inhibits the insulin-like growth factor-1 receptor and anaplastic lymphoma kinase and shows antitumor activity in experimental models of human cancers. Mol. Cancer Ther. 8, 2811-2820 (2009). https://doi.org/10.1158/1535-7163.MCT-09-0423 CrossRefPubMedGoogle Scholar
- 33.B.I. Carr, R. D'Alessandro, M.G. Refolo, P.A. Iacovazzi, C. Lippolis, C. Messa, A. Cavallini, M. Correale, A. Di Carlo, Effects of low concentrations of Regorafenib and Sorafenib on human HCC cell AFP, migration, invasion, and growth in vitro. J. Cell. Physiol. 228, 1344-1350 (2013). https://doi.org/10.1002/jcp.24291 CrossRefPubMedPubMedCentralGoogle Scholar
- 34.T.C. Chou, in Synergism and Antagonism in Chemotherapy, ed. by T. C. Chou, D. C. Rideout. The median-effect principle and the combination index for quantitation of synergism and antagonism (Academic Press, San Diego, 1991), pp. 61–102Google Scholar
- 39.A.H. Rosendahl, C. Gundewar, K. Said Hilmersson, L. Ni, M.A. Saleem, R. Andersson, Conditionally immortalized human pancreatic stellate cell lines demonstrate enhanced proliferation and migration in response to IGF-I. Exp. Cell Res. 330, 300-310 (2015). https://doi.org/10.1016/j.yexcr.2014.09.033 CrossRefPubMedGoogle Scholar
- 46.Y.P. Ching, V.Y. Leong, M.F. Lee, H.T. Xu, D.Y. Jin, I.O. Ng, P21-activated protein kinase is overexpressed in hepatocellular carcinoma and enhances cancer metastasis involving c-Jun NH2-terminal kinase activation and paxillin phosphorylation. Cancer Res. 67, 3601-3608 (2007). https://doi.org/10.1158/0008-5472.CAN-06-3994 CrossRefPubMedGoogle Scholar