Abstract
Purpose
Immunosuppressive therapy after liver transplantation for hepatocellular carcinoma (HCC) is one of the major contributory factors for HCC recurrence and metastasis. Sirolimus, a potent immunosuppressant, has been reported to be an effective inhibitor in a variety of tumors. The present study is designed to explore whether sirolimus could block the growth and metastatic progression of HCC.
Methods
MHCC97H cells were used as targets to explore the effect of sirolimus on cell cycle progression, apoptosis, proliferation, and its antiangiogenic mechanism. LCI-D20, a highly metastatic model of human HCC in nude mice, was also used as the model tumor to explore the effect of sirolimus on tumor growth and metastatic progression.
Results
In vitro, sirolimus induced cell cycle arrest at the G1 checkpoint and blocked proliferation of MHCC97H cells but did not induce apoptosis. In vivo, sirolimus prevented tumor growth and metastatic progression in LCI-D20. Intratumoral microvessel density and circulating levels of VEGF in tumor-bearing mice were also significantly reduced in sirolimus treatment group. Quantitative RT-PCR showed that sirolimus down-regulated the mRNA expression of VEGF and HIF-1a, but not of bFGF, and TGF-b in MHCC97H cells. Furthermore, western blot analysis confirmed that sirolimus also decreased expression of HIF-1a at protein level, in parallel with the down-regulation of the levels of VEGF protein excretion in a time-dependent manner as compared to untreated control cells following anoxia.
Conclusions
The immunosuppressive macrolide sirolimus prevents the growth and metastatic progression of HCC, and suppresses VEGF synthesis and secretion by downregulating HIF-1a expression. Sirolimus may be useful for clinical application in patients who received a liver transplant for HCC.
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Abbreviations
- HCC:
-
Hepatocellular carcinoma
- LT:
-
Liver transplantation
- CNI:
-
Calcineurin inhibitor
- MVD:
-
Microvessel density
References
Blancher C, Moore JW, Robertson N, Harris AL (2001) Effects of ras and von Hippel-Lindau (VHL) gene mutations on hypoxiainducible factor (HIF)-1 alpha, HIF-2 alpha, and vascular endothelial growth factor expression and their regulation by the phosphatidylinositol 3′-kinase/Akt signaling pathway. Cancer Res 61:7349–7355
Boffa DJ, Luan F, Thomas D, Yang H, Sharma VK, Lagman M et al (2004) Rapamycin inhibits the growth and metastatic progression of non-small cell lung cancer. Clin Cancer Res 10:293–300. doi:10.1158/1078-0432.CCR-0629-3
Breier G, Blum S, Peli J, Groot M, Wild C, Risau W et al (2002) Transforming growth factor-beta and Ras regulate the VEGF/VEGF-receptor system during tumor angiogenesis. Int J Cancer 97:142–148. doi:10.1002/ijc.1599
Burke B, Tang N, Corke KP, Tazzyman D, Ameri K, Wells M et al (2002) Expression of HIF-1alpha by human macrophages: implications for the use of macrophages in hypoxiaregulated cancer gene therapy. J Pathol 196:204–212. doi:10.1002/path.1029
Chan S, Scheulen ME, Johnston S, Mross K, Cardoso F, Dittrich C et al (2005) Phase II study of temsirolimus (CCI-779), a novel inhibitor of mTOR, in heavily pretreated patients with locally advanced or metastatic breast cancer. J Clin Oncol 23:5314–5322. doi:10.1200/JCO.2005.66.130
Clarke K, Smith K, Gullick WJ, Harris AL (2001) Mutant epidermal growth factor receptor enhances induction of vascular endothelial growth factor by hypoxia and insulin-like growth factor-1 via a PI3 kinase dependent pathway. Br J Cancer 84:1322–1399. doi:10.1054/bjoc.2001.1805
El-Serag HB, Mason AC (1999) Rising incidence of hepatocellular carcinoma in the United States. N Engl J Med 340:745–750. doi:10.1056/NEJM199903113401001
Freise CE, Ferrell L, Liu T, Ascher NL, Roberts JP (1999) Effect of systemic cyclosporine on tumor recurrence after liver transplantation in a model of hepatocellular carcinoma. Transplantation 67:510–513. doi:10.1097/00007890-199902270-00003
Guba M, von Breitenbuch P, Steinbauer M, Koehl G, Flegel S, Hornung M et al (2002) Rapamycin inhibits primary and metastatic tumor growth by antiangiogenesis: involvement of vascular endothelial growth factor. Nat Med 8:128–135. doi:10.1038/nm0202-128
Hale DA, Gottschalk R, Fukuzaki T, Wood ML, Maki T, Monaco AP (1997) Superiority of sirolimus (rapamycin) over cyclosporine in augmenting allograft and xenograft survival in mice treated with antilymphocyte serum and donor-specific bone marrow. Transplantation 63:359–364. doi:10.1097/00007890-199702150-00005
Hojo M, Morimoto T, Maluccio M, Asano T, Morimoto K, Lagman M et al (1999) Cyclosporine induces cancer progression by a cell-autonomous mechanism. Nature 397:530–534. doi:10.1038/17401
Hudes G, Carducci M, Tomczak P, Dutcher J, Figlin R, Kapoor A, Global ARCC Trial et al (2007) Temsirolimus, interferon alfa, or both for advanced renal-cell carcinoma. N Engl J Med 356:2271–2281. doi:10.1056/NEJMoa066838
Hudson CC, Liu M, Chiang GG, Otterness DM, Loomis DC, Kaper F et al (2002) Regulation of hypoxia-inducible factor 1alpha expression and function by the mammalian target of rapamycin. Mol Cell Biol 22:7004–7014. doi:10.1128/MCB.22.20.7004-7014.2002
Jain RK, Safabakhsh N, Sckell A, Chen Y, Jiang P, Benjamin L et al (1998) Endothelial cell death, angiogenesis, and microvascular function after castration in an androgendependent tumor: role of vascular endothelial growth factor. Proc Natl Acad Sci USA 95:10820–10825. doi:10.1073/pnas.95.18.10820
Kneteman NM, Oberholzer J, Al Saghier M, Meeberg GA, Blitz M, Ma MM et al (2004) Sirolimus-based immunosuppression for liver transplantation in the presence of extended criteria for hepatocellular carcinoma. Liver Transpl 10:1301–1311. doi:10.1002/lt.20237
Luan FL, Hojo M, Maluccio M, Yamaji K, Suthanthiran M (2002) Rapamycin blocks tumor progression: unlinking immunosuppression from antitumor efficacy. Transplantation 73:1565–1572. doi:10.1097/00007890-200205270-00008
Mahalati K, Kahan BD (2001) Clinical pharmacokinetics of sirolimus. Clin Pharmacokinet 40:573–585. doi:10.2165/00003088-200140080-00002
Olthoff KM (1998) Surgical options for hepatocellular carcinoma: resection and transplantation. Liver Transpl Surg 4:S98–S104
Rathmell WK, Wright TM, Rini BI (2005) Molecularly targeted therapy in renal cell carcinoma. Expert Rev Anticancer Ther 5:1031–1040. doi:10.1586/14737140.5.6.1031
Semela D, Piguet AC, Kolev M, Schmitter K, Hlushchuk R, Djonov V et al (2007) Vascular remodeling and antitumoral effects of mTOR inhibition in a rat model of hepatocellular carcinoma. J Hepatol 46:840–848. doi:10.1016/j.jhep.2006.11.021
Sun FX, Tang ZY, Liu KD, Xue Q, Gao DM, Yu YQ et al (1996) Establishment of a metastatic model of human hepatocellular carcinoma in nude mice via orthotopic implantation of histologically intact tissues. Int J Cancer 66:239–243. doi:10.1002/(SICI)1097-0215(19960410)66:2<239::AID-IJC17>3.0.CO;2-7
Thomas MB, Zhu AX (2005) Hepatocellular carcinoma: the need for progress. J Clin Oncol 23:2892–2899. doi:10.1200/JCO.2005.03.196
Toso C, Meeberg GA, Bigam DL (2007) De novo sirolimus-based immunosuppression after liver transplantation for hepatocellular carcinoma: long-term outcomes and side effects. Transplantation 83:1162–1168. doi:10.1097/01.tp.0000262607.95372.e0
Vivarelli M, Cucchetti A, Piscaglia F, La Barba G, Bolondi L, Cavallari A et al (2005) Analysis of risk factors for tumor recurrence after liver transplantation for hepatocellular carcinoma: key role of immunosuppression. Liver Transpl 11:497–503. doi:10.1002/lt.20391
Wiedmann MW, Caca K (2005) Molecularly targeted therapy for gastrointestinal cancer. Curr Cancer Drug Targets 5:171–193. doi:10.2174/1568009053765771
Yang R, Rescorla FJ, Reilly CR, Faught PR, Sanghvi NT, Lumeng L et al (1992) A reproducible rat liver cancer model for experimental therapy: introducing a technique of intrahepatic tumor implantation. J Surg Res 52:193–198. doi:10.1016/0022-4804(92)90072-8
Zhang JF, Liu JJ, Lu MQ, Cai CJ, Yang Y, Li H et al (2007) Rapamycin inhibits cell growth by induction of apoptosis on hepatocellular carcinoma cells in vitro. Transpl Immunol 17:162–168. doi:10.1016/j.trim.2006.12.003
Zhang L, Conejo-Garcia JR, Yang N, Huang W, Mohamed-Hadley A, Yao W et al (2002) Different effects of glucose starvation on expression and stability of VEGF mRNA isoforms in murine ovarian cancer cells. Biochem Biophys Res Commun 292:860–868. doi:10.1006/bbrc.2002.6710
Zimmerman MA, Trotter JF, Wachs M, Bak T, Campsen J, Skibba A et al (2008) Sirolimus-based immunosuppression following liver transplantation for hepatocellular carcinoma. Liver Transpl 14:633–638. doi:10.1002/lt.21420
Acknowledgments
The research was supported by grants from the Hi-tech Research and Development Program of China (2007AA02Z479), and the National Natural Science Foundation Grant of China (30700815).
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Z. Wang and J. Zhou contributed equally to this work.
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Wang, Z., Zhou, J., Fan, J. et al. Sirolimus inhibits the growth and metastatic progression of hepatocellular carcinoma. J Cancer Res Clin Oncol 135, 715–722 (2009). https://doi.org/10.1007/s00432-008-0506-z
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DOI: https://doi.org/10.1007/s00432-008-0506-z