Inflammation Promotes Expression of Stemness-Related Properties in HBV-Related Hepatocellular Carcinoma
Te-Sheng Chang, Chi-Long Chen, Yu-Chih Wu, Jun-Jen Liu, Yung Che Kuo, Kam-Fai Lee, Sin-Yi Lin, Sey-En Lin, Shui-Yi Tung, Liang-Mou Kuo, Ying-Huang Tsai, Yen-Hua Huang, Te-Sheng Chang, Chi-Long Chen, Yu-Chih Wu, Jun-Jen Liu, Yung Che Kuo, Kam-Fai Lee, Sin-Yi Lin, Sey-En Lin, Shui-Yi Tung, Liang-Mou Kuo, Ying-Huang Tsai, Yen-Hua Huang
Abstract
The expression of cancer stemness is believed to reduce the efficacy of current therapies against hepatocellular carcinoma (HCC). Understanding of the stemness-regulating signaling pathways incurred by a specific etiology can facilitate the development of novel targets for individualized therapy against HCC. Niche environments, such as virus-induced inflammation, may play a crucial role. However, the mechanisms linking inflammation and stemness expression in HCC remain unclear. Here we demonstrated the distinct role of inflammatory mediators in expressions of stemness-related properties involving the pluripotent octamer-binding transcription factor 4 (OCT4) in cell migration and drug resistance of hepatitis B virus-related HCC (HBV-HCC). We observed positive immunorecognition for macrophage chemoattractant protein 1 (MCP-1)/CD68 and OCT4/NANOG in HBV-HCC tissues. The inflammation-conditioned medium (inflamed-CM) generated by lipopolysaccharide-stimulated U937 human leukemia cells significantly increased the mRNA and protein levels of OCT4/NANOG preferentially in HBV-active (HBV+HBsAg+) HCC cells. The inflamed-CM also increased the side population (SP) cell percentage, green fluorescent protein (GFP)-positive cell population, and luciferase activity of OCT4 promoter-GFP/luciferase in HBV-active HCC cells. Furthermore, the inflamed-CM upregulated the expressions of insulin-like growth factor-I (IGF-I)/IGF-I receptor (IGF-IR) and activated IGF-IR/Akt signaling in HBV-HCC. The IGF-IR phosphorylation inhibitor picropodophyllin (PPP) suppressed inflamed-CM-induced OCT4 and NANOG levels in HBV+HBsAg+ Hep3B cells. Forced expression of OCT4 significantly increased the secondary sphere formation and cell migration, and reduced susceptibility of HBV-HCC cells to cisplatin, bleomycin, and doxorubicin. Taking together, our results show that niche inflammatory mediators play critical roles in inducing the expression of stemness-related properties involving IGF-IR activation, and the upregulation of OCT4 contributes to cancer migration and drug resistance of HBV-HCC cells. Findings in this paper would provide potential targets for a therapeutic strategy targeting on inflammatory environment for HBV-HCC.
Conflict of interest statement
Competing Interests: The authors have declared that no competing interests exist.
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References
- Coussens LM, Werb Z. Inflammation and cancer. Nature. 2002;420: 860–867.
- Berasain G, Castillo J, Perugorria MJ, Latasa MU, Prieto J, Avila MA. Inflammation and liver cancer. Ann NY Acad Sci. 2009;1155: 206–221. 10.1111/j.1749-6632.2009.03704.x
- Arndt W, Sandra K, Ina N, Henning S, Jochem K, Maria H, et al. Trends in epidemiology, treatment, and survival of hepatocellular carcinoma patients between 1998 and 2009: an analysis of 1066 cases of a German HCC registry. J Clin Gastroenterol. 2014;48: 279–289. 10.1097/MCG.0b013e3182a8a793
- Yang JD, Nakamura I, Roberts LR. The tumor microenvironment in hepatocellular carcinoma: Current status and therapeutic targets. Semin Cancer Biol. 2011;21: 35–43. 10.1016/j.semcancer.2010.10.007
- Ajani JA, Song S, Hochster HS, Steinburg IB. Cancer Stem Cells: The promise and the potential. Semin Oncol. 2015;42: S3–S17.
- Tanno T, Matsui W. Development and maintenance of cancer stem cells under chronic inflammation. J Nippon Med Sch. 2011;78: 138–145.
- Shigdar S, Li Y, Bhattacharya S, O'Connor M, Pu C, Lin J, et al. Inflammation and cancer stem cells. Cancer Lett. 2014;345: 271–278. 10.1016/j.canlet.2013.07.031
- Csermely P, Hodsagi J, Korcsmaros T, Módos D, Perez-Lopez AR, Szalay K, et al. Cancer stem cells display extremely large evolvability: Alternating plastic and rigid networks as a potential mechanism: Network models, novel therapeutic target strategies, and the contributions of hypoxia, inflammation and cellular senescence. Semin Cancer Biol. 2015;30: 42–51. 10.1016/j.semcancer.2013.12.004
- Jinushi M, Chiba S, Yoshiyama H, Masutomi K, Kinoshita I, Dosaka-Akita H, et al. Tumor-associated macrophages regulate tumorigenicity and anticancer drug responses of cancer stem/initiating cells. Proc Natl Acad Sci USA. 2011;108: 12425–12430. 10.1073/pnas.1106645108
- Ma N, Thanan R, Kobayashi H, Hamman O, Wishahi M, Leithy TE, et al. Nitrative DNA damage and Oct3/4 expression in urinary bladder cancer with Schistosoma haematobium infection. Biochem Bioph Res Co. 2011;414: 344–349.
- Knight B, Matthews VB, Akhurst B, Croager EJ, Klinken E, Abraham LJ, et al. Liver inflammation and cytokine production, but not acute phase protein synthesis accompany the adult liver progenitor (oval) cell response to chronic liver injury. Immunol Cell Biol. 2005;83: 364–374.
- He G, Dhar D, Nakagawa H, Font-Burgada J, Ogata H, Jiang Y, et al. Identification of liver cancer progenitors whose malignant progression depends on autocrine IL-6 signaling. Cell. 2013;155: 384–396. 10.1016/j.cell.2013.09.031
- Boyault S, Rickman DS, de Reyniès A, Balabaud C, Rebouissou S, Jeannot E, et al. Transcriptome classification of HCC is related to gene alterations and to new therapeutic targets. Hepatology. 2007;45: 42–52.
- Chang TS, Wu YC, Chi CC, Su WC, Chang PJ, Lee KF, et al. Activation of IL6/IGF-IR confers poor prognosis of HBV-related hepatocellular carcinoma through induction of OCT4/NANOG expression. Clin Cancer Res. 2015;21: 201–210. 10.1158/1078-0432.CCR-13-3274
- Bendall SC, Stewart MH, Menendez P. George D, Vijayaragavan K, Werbowetski-Ogilvie T, et al. IGF and FGF cooperatively establish the regulatory stem cell niche of pluripotent human cells in vitro. Nature. 2007; 448: 1015–1023.
- Wang L, Schulz TC, Sherrer ES, Dauphin DS, Shin S, Nelson AM, et al. Self-renewal of human embryonic stem cells requires insulin-like growth factor-1 receptor and ERBB2 receptor signaling. Blood. 2007;110: 4111–4119.
- Huang YH, Chin CC, Ho HN, Chou CK, Shen CN, Kuo HC, et al. Pluripotency of mouse spermatogonial stem cells maintained by IGF-I-dependent pathway. FASEB J. 2009;23: 2076–2087. 10.1096/fj.08-121939
- Wang XQ, Ongkeko WM, Chen L, Yang ZF, Lu P, Chen KK, et al. Octomer 4 (Oct4) mediates chemotherapeutic drug resistance in liver cancer cells through a potential Oct4–AKT–ATP-binding cassette G2 pathway. Hepatology. 2010;52: 528–539. 10.1002/hep.23692
- Desbois-Mouthon C, Baron A, Eggelpoel MB, Fartoux L, Venot C, Bladt F, et al. Insulin-like growth factor-1 receptor inhibition induces a resistance mechanism via the epidermal growth factor receptor/HER3/AKT signaling pathway: rational basis for cotargeting insulin-like growth factor-1 receptor and epidermal growth factor receptor in hepatocellular carcinoma. Clin Cancer Res. 2009;15: 5445–5456. 10.1158/1078-0432.CCR-08-2980
- Ramanan V, Scull MA, Sheahan TP, Rice CM, Bhatia SN. New methods in tissue engineering: Improved models for viral infection. Annu Rev Virol. 2014;1: 475–499.
- Chiao H, Foster S, Thomas R, Lipton J, Star RA. Alpha-melanocyte-stimulating hormone reduces endotoxin-induced liver inflammation. J Clin Invest. 1996;97: 2038–2044.
- Tarao K, Takemiya S, Tamai S, Sugimasa Y, Ohkawa S, Akaike M, et al. Relationship between the recurrence of hepatocellular carcinoma (HCC) and serum alanine aminotransferase levels in hepatectomized patients with hepatitis C virus-associated cirrhosis and HCC. Cancer. 1997;79: 688–694.
- Matsumoto K, Yoshimoto J, Sugo H, Kojima K, Futagawa S, Matsumoto T. Relationship between the histological degrees of hepatitis and the postoperative recurrence of hepatocellular carcinoma in patients with hepatitis C. Hepatol Res. 2002;23: 196–201.
- Cescon M, Cucchetti A, Grazi GL, Ferrero A, Viganò L, Ercolani G, et al. Role of hepatitis B virus infection in the prognosis after hepatectomy for hepatocellular carcinoma in patients with cirrhosis. Arch Surg. 2009;144: 906–913. 10.1001/archsurg.2009.99
- Wu HC, Tsai HW, Teng CF, Hsieh WC, Lin YJ, Wang HC, et al. Ground-glass hepatocytes co-expressing hepatitis B virus X protein and surface antigens exhibit enhanced oncogenic effects and tumorigenesis. Hum Pathol. 2014;45: 1294–1301. 10.1016/j.humpath.2013.10.039
- Sell S, Leffert HL. Liver cancer stem cells. J Clin Oncol. 2008;26: 2800–2805. 10.1200/JCO.2007.15.5945
- Takahashi K, Yamanaka S. Induction of pluripotent stem cells from mouse embryonic and adult fibroblast cultures by defined factors. Cell. 2006:126: 663–676.
- Shan J, Shen J, Liu L, Xia F, Xu C, Duan G, et al. Nanog regulates self-renewal of cancer stem cells through the insulin-like growth factor pathway in human hepatocellular carcinoma. Hepatology. 2012;56: 1004–1014. 10.1002/hep.25745
- Chiba T, Kita K, Zheng YW, Yokosuka O, Saisho H, Iwama A, et al. Side population purified from hepatocellular carcinoma cells harbors cancer stem cell–like properties. Hepatology. 2012;44: 240–251.
- Tovar V, Alsinet C, Villanueva A, Hoshida Y, Chiang DY, Solé M, et al. IGF activation in a molecular subclass of hepatocellular carcinoma and pre-clinical efficacy of IGF-IR blockage. J Hepatol. 2010;52: 550–559. 10.1016/j.jhep.2010.01.015
- Llovet JM, Ricci S, Mazzaferro V, Hilgard P, Gane E, Blanc JF, et al. Sorafenib in advanced hepatocellular carcinoma. N Engl J Med. 2008;359: 378–390. 10.1056/NEJMoa0708857
- Fransvea E, Paradiso A, Antonaci S, Giannelli G. HCC heterogeneity: molecular pathogenesis and clinical implications. Cell Oncol. 2009;31: 227–233. 10.3233/CLO-2009-0473
- Villanueva A, Newell P, Chiang DY, Friedman SL, Llovet JM. Genomics and signaling pathways in hepatocellular carcinoma. Semin Liver Dis. 2007;27: 55–76.
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