Abstract
Purpose
Deoxycholic acid (DCA), a secondary bile acid, is reportedly increased in the serum of patients with nonalcoholic steatohepatitis and animals with experimentally induced hepatocellular carcinoma (HCC), but its contribution to malignant behaviors of HCC has not been precisely clarified. This study aimed to examine the effect of DCA on hepatic stellate cells (HSCs), a major component of nonparenchymal cells in the liver, and its subsequent indirect effect on HCC cells.
Methods
LX2 cells, a human HSC line, were treated with DCA in vitro. Then, HuH7 cells, a human hepatoma cell line, were incubated in conditioned media of DCA-treated LX2 to investigate the subsequent effect focusing on malignant behaviors.
Results
DCA resulted in cellular senescence in LX2 with the decreased cell proliferation via cell cycle arrest at G0/1 phase, together with the induction of senescence-associated secretory phenotype (SASP) factors. To investigate the influence of SASP factors secreted by HSCs in response to DCA, HCC cells were treated with conditioned media that promoted cell migration and invasion via induction of epithelial mesenchymal transition. These changes were attenuated in the presence of neutralizing antibody against IL8 or TGFβ. Pathological analysis of surgical specimens from HCC patients revealed that senescent HSCs were detected in the stroma surrounding HCC.
Conclusion
Our data suggest an important role of HSC senescence caused by DCA for the malignant biological behaviors of HCC via induction of SASP factors, particularly IL8 and TGFβ.
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References
Aranha MM, Cortez-Pinto H, Costa A, da Silva IB, Camilo ME, de Moura MC, Rodrigues CM (2008) Bile acid levels are increased in the liver of patients with steatohepatitis. Eur J Gastroenterol Hepatol 20:519–525. https://doi.org/10.1097/MEG.0b013e3282f4710a
Ascha MS, Hanouneh IA, Lopez R, Tamimi TA, Feldstein AF, Zein NN (2010) The incidence and risk factors of hepatocellular carcinoma in patients with nonalcoholic steatohepatitis. Hepatology 51:1972–1978. https://doi.org/10.1002/hep.23527
Bilusic M et al (2019) Phase I trial of HuMax-IL8 (BMS-986253), an anti-IL-8 monoclonal antibody, in patients with metastatic or unresectable solid tumors. J Immunother Cancer. https://doi.org/10.1186/s40425-019-0706-x
Bray F, Ferlay J, Soerjomataram I, Siegel RL, Torre LA, Jemal A (2018) Global cancer statistics 2018: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries. CA Cancer J Clin 68:394–424. https://doi.org/10.3322/caac.21492
Campisi J (2005) Senescent cells, tumor suppression, and organismal aging: good citizens, bad neighbors. Cell 120:513–522. https://doi.org/10.1016/j.cell.2005.02.003
Cook JW, Kennaway EI, Kennaway NM (1940) Production of tumours in mice by deoxycholic acid. Nature 145:627–627. https://doi.org/10.1038/145627a0
Coppe JP et al (2008) Senescence-associated secretory phenotypes reveal cell-nonautonomous functions of oncogenic RAS and the p53 tumor suppressor. PLoS Biol 6:2853–2868. https://doi.org/10.1371/journal.pbio.0060301
d'Adda di Fagagna F (2008) Living on a break: cellular senescence as a DNA-damage response. Nat Rev Cancer 8:512–522. https://doi.org/10.1038/nrc2440
Evangelou K, Gorgoulis VG (2017) Sudan Black B, the specific histochemical stain for lipofuscin: a novel method to detect senescent cells. Methods Mol Biol 1534:111–119. https://doi.org/10.1007/978-1-4939-6670-7_10
Fabregat I, Roncero C, Fernandez M (2007) Survival and apoptosis: a dysregulated balance in liver cancer. Liver Int 27:155–162. https://doi.org/10.1111/j.1478-3231.2006.01409.x
Frey N, Venturelli S, Zender L, Bitzer M (2018) Cellular senescence in gastrointestinal diseases: from pathogenesis to therapeutics. Nat Rev Gastroenterol Hepatol 15:81–95. https://doi.org/10.1038/nrgastro.2017.146
Friedman SL (2008) Hepatic stellate cells: protean, multifunctional, and enigmatic cells of the liver. Physiol Rev 88:125–172. https://doi.org/10.1152/physrev.00013.2007
Fu XT et al (2015) Macrophage-secreted IL-8 induces epithelial-mesenchymal transition in hepatocellular carcinoma cells by activating the JAK2/STAT3/Snail pathway. Int J Oncol 46:587–596. https://doi.org/10.3892/ijo.2014.2761
Guo M et al (2013) The role of vimentin intermediate filaments in cortical and cytoplasmic mechanics. Biophys J 105:1562–1568. https://doi.org/10.1016/j.bpj.2013.08.037
Honma N, Genda T, Matsuda Y, Yamagiwa S, Takamura M, Ichida T, Aoyagi Y (2006) MEK/ERK signaling is a critical mediator for integrin-induced cell scattering in highly metastatic hepatocellular carcinoma cells. Lab Invest 86:687–696. https://doi.org/10.1038/labinvest.3700427
Huang WD et al (2006) Nuclear receptor-dependent bile acid signaling is required for normal liver regeneration. Science 312:233–236. https://doi.org/10.1126/science.1121435
Hustedt N, Durocher D (2016) The control of DNA repair by the cell cycle. Nat Cell Biol 19:1–9. https://doi.org/10.1038/ncb3452
Jemal A et al (2017) Annual report to the nation on the status of cancer, 1975–2014. Featuring Survival J Natl Cancer Inst. https://doi.org/10.1093/jnci/djx030
Karimian A, Ahmadi Y, Yousefi B (2016) Multiple functions of p21 in cell cycle, apoptosis and transcriptional regulation after DNA damage. DNA Repair (Amst) 42:63–71. https://doi.org/10.1016/j.dnarep.2016.04.008
Kaul Z, Cesare AJ, Huschtscha LI, Neumann AA, Reddel RR (2012) Five dysfunctional telomeres predict onset of senescence in human cells. Embo Rep 13:52–59. https://doi.org/10.1038/embor.2011.227
Kim JH (2020) Interleukin-8 in the tumor immune niche: lessons from comparative oncology. Adv Exp Med Biol 1240:25–33. https://doi.org/10.1007/978-3-030-38315-2_2
Kim I, Morimura K, Shah Y, Yang Q, Ward JM, Gonzalez FJ (2007) Spontaneous hepatocarcinogenesis in farnesoid X receptor-null mice vol 28. Carcinogenesis. https://doi.org/10.1093/carcin/bgl249
Knisely AS et al (2006) Hepatocellular carcinoma in ten children under 5 years of age with bile salt export pump deficiency. Hepatology 44:478–486. https://doi.org/10.1002/hep.21287
Kuilman T et al (2008) Oncogene-induced senescence relayed by an interleukin-dependent inflammatory network. Cell 133:1019–1031. https://doi.org/10.1016/j.cell.2008.03.039
Liu H et al (2015) Ursodeoxycholic acid induces apoptosis in hepatocellular carcinoma xenografts in mice. World J Gastroenterol 21:10367–10374. https://doi.org/10.3748/wjg.v21.i36.10367
Llovet JM, Bruix J (2008) Molecular targeted therapies in hepatocellular carcinoma. Hepatology 48:1312–1327. https://doi.org/10.1002/hep.22506
Lowe SW, Cepero E, Evan G (2004) Intrinsic tumour suppression. Nature 432:307–315. https://doi.org/10.1038/nature03098
Mannaerts I, Schroyen B, Verhulst S, Van Lommel L, Schuit F, Nyssen M, van Grunsven LA (2013) Gene expression profiling of early hepatic stellate cell activation reveals a role for Igfbp3 in cell migration. PLoS ONE 8:e84071. https://doi.org/10.1371/journal.pone.0084071
Mariotti A, Perotti A, Sessa C, Ruegg C (2007) N-cadherin as a therapeutic target in cancer Expert. Opin Inv Drug 16:451–465. https://doi.org/10.1517/13543784.16.4.451
Mikula M, Proell V, Fischer AN, Mikulits W (2006) Activated hepatic stellate cells induce tumor progression of neoplastic hepatocytes in a TGF-beta dependent fashion. J Cell Physiol 209:560–567. https://doi.org/10.1002/jcp.20772
Nguyen PT, Kudo Y, Yoshida M, Iizuka S, Ogawa I, Takata T (2011) N-cadherin expression is correlated with metastasis of spindle cell carcinoma of head and neck region. J Oral Pathol Med 40:77–82. https://doi.org/10.1111/j.1600-0714.2010.00966.x
Nguyen PT, Tsunematsu T, Yanagisawa S, Kudo Y, Miyauchi M, Kamata N, Takata T (2013) The FGFR1 inhibitor PD173074 induces mesenchymal-epithelial transition through the transcription factor AP-1. Brit J Cancer 109:2248–2258. https://doi.org/10.1038/bjc.2013.550
Nurse P (2000) A long twentieth century of the cell cycle and beyond. Cell 100:71–78. https://doi.org/10.1016/S0092-8674(00)81684-0
Oda K, Uto H, Mawatari S, Ido A (2015) Clinical features of hepatocellular carcinoma associated with nonalcoholic fatty liver disease: a review of human studies Clin. J Gastroenterol 8:1–9. https://doi.org/10.1007/s12328-014-0548-5
Saga K et al (2018) Secondary unconjugated bile acids induce hepatic stellate cell activation. Int J Mol Sci. https://doi.org/10.3390/ijms19103043
Son H, Moon A (2010) Epithelial-mesenchymal transition and cell invasion. Toxicol Res 26:245–252. https://doi.org/10.5487/TR.2010.26.4.245
Sun L et al (2016) Bile acids promote diethylnitrosamine-induced hepatocellular carcinoma via increased inflammatory signaling. Am J Physiol Gastrointest Liver Physiol 311:G91–G104. https://doi.org/10.1152/ajpgi.00027.2015
Taura K et al (2008) Hepatic stellate cells secrete angiopoietin 1 that induces angiogenesis in liver fibrosis. Gastroenterology 135:1729–1738. https://doi.org/10.1053/j.gastro.2008.07.065
van Zijl F et al (2009) Hepatic tumor-stroma crosstalk guides epithelial to mesenchymal transition at the tumor edge. Oncogene 28:4022–4033. https://doi.org/10.1038/onc.2009.253
Xing S et al (2018) Isoviolanthin extracted from dendrobium officinale reverses TGF-beta1-mediated epithelial(-)mesenchymal transition in hepatocellular carcinoma cells via deactivating the TGF-beta/Smad and PI3K/Akt/mTOR signaling pathways. Int J Mol Sci. https://doi.org/10.3390/ijms19061556
Xu J, Lamouille S, Derynck R (2009) TGF-beta-induced epithelial to mesenchymal transition. Cell Res 19:156–172. https://doi.org/10.1038/cr.2009.5
Yamada S, Takashina Y, Watanabe M, Nagamine R, Saito Y, Kamada N, Saito H (2018) Bile acid metabolism regulated by the gut microbiota promotes non-alcoholic steatohepatitis-associated hepatocellular carcinoma in mice. Oncotarget 9:9925–9939. https://doi.org/10.18632/oncotarget.24066
Yin C, Evason KJ, Asahina K, Stainier DY (2013) Hepatic stellate cells in liver development, regeneration, and cancer. J Clin Invest 123:1902–1910. https://doi.org/10.1172/JCI66369
Yoshimoto S et al (2013) Obesity-induced gut microbial metabolite promotes liver cancer through senescence secretome. Nature 499:97–101. https://doi.org/10.1038/nature12347
Acknowledgements
This work was supported in part by grants-in-aid from the Japan Society for the Promotion of Science to P.T. Nguyen, and K. Kanno. We are grateful the great support from Dr. Akihiro Kawahara and Sayaka Yonezawa for this study. The results here are in part based upon data generated by the TCGA Research Network: https://www.cancer.gov/tcga.
Funding
This work was supported in part by grants-in-aid from the Japan Society for the Promotion of Science to P.T. Nguyen and K. Kanno.
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PTN, ST, NK, and KK conceived and planned the experiments. PTN and QTP carried out the experiments. MM and KA contributed to sample preparation. YK, MK, TK, TO, and MI contributed to the interpretation of the results. PTN and KK took the lead in writing the manuscript.
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Nguyen, P.T., Kanno, K., Pham, Q.T. et al. Senescent hepatic stellate cells caused by deoxycholic acid modulates malignant behavior of hepatocellular carcinoma. J Cancer Res Clin Oncol 146, 3255–3268 (2020). https://doi.org/10.1007/s00432-020-03374-9
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DOI: https://doi.org/10.1007/s00432-020-03374-9