NFκB activation demarcates a subset of hepatocellular carcinoma patients for targeted therapy
- 532 Downloads
Hepatocellular carcinoma (HCC) is the fifth most common cancer and the third leading cause of cancer death worldwide. It is a heterogeneous disorder and >80 % of the tumors develop in patients with liver cirrhosis, resulting from chronic inflammation and/or fibrosis. Here, we set out to identify novel targets for HCC therapy and to define a subgroup of patients that might benefit most from it.
Cellular pathway activation profiling of 45 transcription factors in a HCC-derived cell line (HEP3B), in vitro analysis of NFκB reporter activity in additional HCC-derived cell lines and pathway-focused integrative analyses of publicly available primary HCC-derived expression profiling data (GSE6764, GSE9843, E-TABM-36 and E-TABM-292) were employed to reveal a role of NFκB in HCC development. In order to identify potential targeting agents, a luciferase-based NFκB reporter screening assay was established in HEP3B cells. After screening of a drug library through this assay, a potent NFκB pathway inhibitor was identified and characterized using an array of additional in vitro assays.
Using cellular pathway activation profiling, we found a high activation of NFκB-mediated signaling in HCC-derived cell lines and in primary HCC tumors. Through NFκB inhibitor screening we observed a highly efficacious NFκB pathway inhibitory potential of ornithogalum in HCC-derived HEP3B cells. Although its active component still remains to be defined, ornithogalum has been found to inhibit endoplasmic reticulum (ER) and oxidative stress responses. ER stress, oxidative stress and NFκB signaling were found to be enhanced in a subset of HCCs, as well as in (precancerous) liver cirrhosis tissues.
From our data we conclude that NFκB signaling is activated in precancerous cirrhosis tissues and in a subset of HCCs. We found that ornithogalum exhibits NFκB targeting and stress relieving activities. NFκB inhibitors, including the active component of ornithogalum, may serve as putative preventive and targeted therapeutic agents for at least a subset of HCCs in which the NFκB pathway is activated. These latter notions require further investigation in a translational context.
KeywordsHepatocellular carcinoma NFkB pathway Targeted therapy Ornithogalum ER stress Oxidative stress Anti-inflammatory
This work was supported by the Department of Atomic Energy, Government of India, through research grant No. 6/6/2008/R&D-II-230R and the Department of Biotechnology, Government of India through research grant BT/PR4500/PID/6/676/2012 to Dr. Kumaresan Ganesan, Madurai Kamaraj University. We acknowledge the award of CSIR-NET fellowship to Vignesh Ramesh. Instrumentation support of the UGC-CEGS, UGC-CAS, DBT-IPLS, DST-PURSE and UGC-NRCBS program and the central facilities of the School of Biological Sciences, Madurai Kamaraj University, are also acknowledged. The authors thank Dr. Piyush Trivedi for providing the drug screening library and Mrs. Jaishree, Publication Division, IIT-Madras, for editorial assistance.
Compliance with ethical standards
Conflict of interest
The authors declare that they have no conflict of interest
- 2.L. Zender, M. S. Spector, W. Xue, P. Flemming, C. Cordon-Cardo, J. Silke, S. T. Fan, J. M. Luk, M. Wigler, G. J. Hannon, D. Mu, R. Lucito, S. Powers, S. W. Lowe, Identification and validation of oncogenes in liver cancer using an integrative oncogenomic approach. Cell 125, 1253–1267 (2006)CrossRefPubMedPubMedCentralGoogle Scholar
- 6.S. Fatima, N. P. Lee, J. M. Luk, Dickkopfs and Wnt/β-catenin signalling in liver cancer. World. J. Clin. Oncol. 2, 311–325 (2011)Google Scholar
- 8.Y. Totoki, K. Tatsuno, S. Yamamoto, Y. Arai, F. Hosoda, S. Ishikawa, S. Tsutsumi, K. Sonoda, H. Totsuka, T. Shirakihara, H. Sakamoto, L. Wang, H. Ojimo, K. Shimada, T. Kosuge, T. Okusaka, K. Kato, J. Kusuda, T. Yoshida, H. Aburatani, T. Shibata, High-resolution characterization of a hepatocellular carcinoma genome. Nat. Genet. 43, 464–469 (2011)CrossRefPubMedGoogle Scholar
- 15.L. Fredriksson, B. Herpers, G. Benedetti, Q. Matadin, J. C. Puigvert, H. de Bont, S. Dragovic, N. P. Vermeulen, J. N. Commandeur, E. Danen, M. de Graauw, B. van de Water, Diclofenac inhibits tumor necrosis factor-α-induced nuclear factor-κB activation causing synergistic hepatocyte apoptosis. Hepatology 53, 2027–2041 (2011)CrossRefPubMedGoogle Scholar
- 16.S. C. Miller, R. Huang, S. Sakamuru, S. J. Shukla, M. S. Attene-Ramos, P. Shinn, D. Van Leer, W. Leister, C. P. Austin, M. Xia, Identification of known drugs that act as inhibitors of NF-kappaB signaling and their mechanism of action. Biochem. Pharmacol. 79, 1272–1280 (2010)CrossRefPubMedPubMedCentralGoogle Scholar
- 17.S. Shishodia, D. Koul, B. B. Aggarwal, Cyclooxygenase (COX)-2 inhibitor celecoxib abrogates TNF-induced NF-kappa B activation through inhibition of activation of I kappa B alpha kinase and Akt in human non-small cell lung carcinoma: correlation with suppression of COX-2 synthesis. J. Immunol. 173, 2011–2022 (2004)CrossRefPubMedGoogle Scholar
- 21.P. N. Verma, I. Vaid, Encyclopaedia of Homeopathic Pharmacopoeia, Vol. IB (Jain Publishers, New Delhi, India, 2002), p. 1864Google Scholar
- 24.A. Subramanian, P. Tamayo, V. K. Mootha, S. Mukherjee, B. L. Ebert, M. A. Gillette, A. Paulovich, S. L. Pomeroy, T. R. Golub, E. S. Lander, J. P. Mesirov, Gene set enrichment analysis: a knowledge-based approach for interpreting genome-wide expression profiles. Proc. Natl. Acad. Sci. U. S. A. 102, 15545–15550 (2005)CrossRefPubMedPubMedCentralGoogle Scholar
- 25.J. Lamb, E. D. Crawford, D. Peck, J. W. Modell, I. C. Blat, M. J. Wrobel, J. Lerner, J. P. Brunet, A. Subramanian, K. N. Ross, M. Reich, H. Hieronymus, G. Wei, S. A. Armstrong, S. J. Haggarty, P. A. Clemons, R. Wei, S. A. Carr, E. S. Lander, T. R. Golub, The connectivity map: using gene-expression signatures to connect small molecules, genes, and disease. Science 313, 1929–1935 (2006)CrossRefPubMedGoogle Scholar
- 32.S. Lüth, J. Schrader, S. Zander, A. Carambia, J. Buchkremer, S. Huber, K. Reifenberg, K. Yamamura, P. Schirmacher, A. W. Lohse, J. Herkel, Chronic inflammatory IFN-γ signaling suppresses hepatocarcinogenesis in mice by sensitizing hepatocytes for apoptosis. Cancer Res. 71, 3763–3771 (2011)CrossRefPubMedGoogle Scholar
- 33.J. F. Rossi, S. Negrier, N. D. James, I. Kocak, R. Hawkins, H. Davis, U. Prabhakar, X. Qin, P. Mulders, B. Berns, A phase I/II study of siltuximab (CNTO 328), an anti-interleukin-6 monoclonal antibody, in metastatic renal cell cancer. Br. J. Cancer 103, 1154–1162 (2010)CrossRefPubMedPubMedCentralGoogle Scholar
- 37.W. Yeo, T. S. Mok, B. Zee, T. W. Leung, P. B. Lai, W. Y. Lau, J. Koh, F. K. Mo, S. C. Yu, A. T. Chan, P. Hui, B. Ma, K. C. Lam, W. M. Ho, H. T. Wong, A. Tang, P. J. Johnson, A randomized phase III study of doxorubicin versus cisplatin/interferon α-2b/doxorubicin/fluorouracil (PIAF) combination chemotherapy for unresectable hepatocellular carcinoma. J. Natl. Cancer Inst. 97, 1532–1538 (2005)CrossRefPubMedGoogle Scholar
- 39.Y. Zhou, C. Garcia-Prieto, D. A. Carney, R. H. Xu, H. Pelicano, Y. Kang, W. Yu, C. Lou, S. Kondo, J. Liu, D. M. Harris, Z. Estrov, M. J. Keating, Z. Jin, P. Huang, OSW-1: a natural compound with potent anticancer activity and a novel mechanism of action. J. Natl. Cancer Inst. 97, 1781–1785 (2005)CrossRefPubMedGoogle Scholar
- 44.M. Shuda, N. Kondoh, N. Imazeki, K. Tanaka, T. Okada, K. Mori, A. Hada, M. Arai, T. Wakatsuki, O. Matsubara, N. Yamamoto, M. Yamamoto, Activation of the ATF6, XBP1 and grp78 genes in human hepatocellular carcinoma: a possible involvement of the ER stress pathway in hepatocarcinogenesis. J. Hepatol. 38, 605–614 (2003)CrossRefPubMedGoogle Scholar
- 47.J. Jin, X. Jin, C. Qian, Y. Ruan, H. Jiang, Signaling network of OSW-1-induced apoptosis and necroptosis in hepatocellular carcinoma. Mol. Med. Rep. 7, 1646–1650 (2013)Google Scholar
- 49.J. M. Luk, C. T. Lam, A. F. Siu, B. Y. Lam, I. O. Ng, M. Y. Hu, C. M. Che, S. T. Fan, Proteomic profiling of hepatocellular carcinoma in Chinese cohort reveals heat-shock proteins (Hsp27, Hsp70, GRP78) up-regulation and their associated prognostic values. Proteomics 6, 1049–1057 (2006)CrossRefPubMedGoogle Scholar