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Cooperation of deregulated Notch signaling and Ras pathway in human hepatocarcinogenesis

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Abstract

Aberrant Notch signaling and Ras pathway had been highlighted a potential role for in human cancers. Yet, relatively little was known about the roles of wild type Notch signaling and Ras in human hepatocarcinogenesis. The aim of this study was to investigate the roles of Ras-Notch signaling cooperation in hepatic cells transformation and proliferation. Hepatocellular carcinoma specimens from 25 patients were analyzed for Notch-1, Ras and Late Simian Virus 40 Factor (LSF) expression using immunohistochemistry. Results showed that Notch-1(76%, 19/25, P < 0.0001), Ras (40%, 10/25, P < 0.01) and LSF (84%, 21/25, P < 0.0001) were significantly up-regulated in hepatocellular carcinoma compared with non-cancer samples. The correlations between the expression and the biological effects of Notch1 and Ras were analyzed by genetic and pharmacological methods. Constitutively active Notch1 alone failed to transform immortalized L02 cells in vivo, it synergized with the Ras pathway to promote hepatic cells transformation. However, their cooperation increased the levels of LSF mRNA and protein, which stimulates L02 cells proliferation. These results exhibited highly aggressive progression, suggesting that Notch-Ras cooperation maybe lead to poor prognosis. Thus, combining the inhibition of the two pathways provided an attractive avenue for therapeutic intervention to overcome this advanced disease.

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Abbreviations

DAPT:

N-(N-[3,5-difluorophenacetyl]-L-alanyl)-Sphenylglycine t-butyl ester

References

  • Bruix J, Sherman M, Practice Guidelines Committee (2005) American association for the study of liver diseases management of hepatocellular carcinoma. Hepatology 42(5):1208–1236. doi:10.1002/hep.20933

    Article  PubMed  Google Scholar 

  • Daniel B, Rangarajan A, Mukherjee G, Vallikad E, Krishna S (1997) The link between integration and expression of human papillomavirus type 16 genomes and cellular changes in the evolution of cervical intraepithelial neoplastic lesions. J Gen Virol 78(Pt5):1095–1101

    PubMed  CAS  Google Scholar 

  • Deftos ML, He YW, Ojata EW, Bevan MJ (1998) Correlating Notch signaling with thymocyte maturation. Immunity 9(6):777–786

    Article  PubMed  CAS  Google Scholar 

  • Donnem T, Andersen S, Al-Shibli K, Al-Saad S, Busund LT, Bremnes RM (2010) Prognostic impact of Notch ligands and receptors in nonsmall cell lung cancer: coexpression of Notch-1 and vascular endothelial growth factor-a predicts poor survival. Cancer 116(24):5676–5685. doi:10.1002/cncr.25551

    Article  PubMed  Google Scholar 

  • Ferrari-Toninelli G, Bonini SA, Uberti D, Buizza L, Bettinsoli P, Poliani PL, Facchetti F, Memo M (2010) Targeting Notch pathway induces growth inhibition and differentiation of neuroblastoma cells. Neuro Oncol 12(12):1231–1243. doi:10.1093/neuonc/noq101

    PubMed  CAS  Google Scholar 

  • Gao J, Chen Y, Wu KC, Liu J, Zhao YQ, Pan YL, Du R, Zheng GR, Xiong YM, Xu HL, Fan DM (2010) RUNX3 directly interacts with intracellular domain of Notch1 and suppresses Notch signaling in hepatocellular carcinoma cells. Exp Cell Res 316(2):149–157. doi:10.1016/j.yexcr.2009.09.025

    Article  PubMed  CAS  Google Scholar 

  • Giovannini C, Gramantieri L, Chieco P, Minguzzi M, Lago F, Pianetti S, Ramazzotti E, Marcu KB, Bolondi L (2009) Selective ablation of Notch3 in HCC enhances doxorubicin’s death promoting effect by a p53 dependent mechanism. J Hepatol 50(5):969–979

    Article  PubMed  CAS  Google Scholar 

  • Gramantieri L, Giovannini C, Lanz A, Chieco P, Ravaioli M, Venturi A, Grazi GL, Bolondi L (2007) Aberrant Notch3 and Notch4 expression in human hepatocellular carcinoma. Liver Int 27(7):997–1007. doi:10.1111/j.1478-3231.2007.01544.x

    Article  PubMed  CAS  Google Scholar 

  • Grieco A, Pompili M, Caminiti G, Miele L, Covino M, Alfei B (2005) Prognostic factors for survival in patients with early intermediate hepatocellular carcinoma undergoing non-surgical therapy: comparison of Okuda, CLIP, and BCLC staging systems in a single Italian centre. Gut 54(3):411–418. doi:10.1136/gut.2004.048124

    Article  PubMed  CAS  Google Scholar 

  • Hansen U, Owens L, Saxena UH (2009) Transcription factors LSF and E2Fs: tandem cyclists driving G0 to S. Cell Cycle 8(14):2146–2151. doi:10.4161/cc.8.14.9089

    Article  PubMed  CAS  Google Scholar 

  • Jain S, Singhal S, Lee P, Xu R (2010) Molecular genetics of hepatocellular neoplasia. Am J Transl Res 2(1):105–118

    PubMed  CAS  Google Scholar 

  • Jehn BM, Bielke W, Pear WS, Osborne BA (1999) Protective effects of Notch-1 on TCR-induced apoptosis. J Immunol 162(2):635–638

    PubMed  CAS  Google Scholar 

  • Jemal A, Siegel R, Xu J, Ward E (2010) Cancer statistics. CA Cancer J Clin 60(5):277–300. doi:10.3322/caac.20073

    Article  PubMed  Google Scholar 

  • Jundt F, Anagnostopoulos I, Forster R, Mathas S, Stein H, Dorken B (2002) Activated Notch1 signaling promotes tumor cell proliferation and survival in Hodgkin and anaplastic large cell lymphoma. Blood 99(9):3398–3403

    Article  PubMed  CAS  Google Scholar 

  • Kimura K, Satoh K, Kanno A, Hamada S, Hirota M, Endoh M, Masamune A, Shimosegawa T (2007) Activation of Notch signaling in tumorigenesis of experimental pancreatic cancer induced by dimethylbenzanthracene in mice. Cancer Sci 98(2):155–162. doi:10.1111/j.1349-7006.2006.00369.x

    Article  PubMed  CAS  Google Scholar 

  • Koch U, Radtke F (2007) Notch and cancer: a double edged sword. Cell Mol Life Sci 64(21):2746–2762. doi:10.1007/s00018-007-7164-1

    Article  PubMed  CAS  Google Scholar 

  • Kubota Y, O’Grady P, Saito H, Takekawa M (2011) Oncogenic Ras abrogates MEK SUMOylation that suppresses the ERK pathway and cell transformation. Nat Cell Biol 13(3):282–291. doi:10.1038/ncb2169

    Article  PubMed  CAS  Google Scholar 

  • Leethanakul C, Patel V, Gillespie J, Pallente M, Ensley JF, Koontongkaew S, Liotta LA, Emmert-Buck M, Gutkind JS (2000) Distinct pattern of expression of differentiation and growth related genes in squamous cell carcinomas of the head and neck revealed by the use of laser capture microdissection and cDNA arrays. Oncogene 19(28):3220–3224

    Article  PubMed  CAS  Google Scholar 

  • Lewis HD, Leveridge M, Strack PR, Haldon CD, O’neil J, Kim H, Madin A, Hannam JC, Look AT, Kohl N, Draetta G, Harrison T, Kerby JA, Shearman MS, Beher D (2007) Apoptosis in T cell acute lymphoblastic leukemia cells after cell cycle arrest induced by pharmacological inhibition of Notch signaling. Chem Biol 14(2):209–219. doi:10.1016/j.chembiol.2006.12.010

    Article  PubMed  CAS  Google Scholar 

  • Malaney S, Daly RJ (2001) The Ras signaling pathway in mammary tumorigenesis and metastasis. J Mammary Gland Biol Neoplasia 6(1):101–113. doi:10.1023/A:1009572700317

    Article  PubMed  CAS  Google Scholar 

  • Masuda S, Kumano K, Suzuki T, Tomita T, Iwatsubo T, Natsugari H, Tojo A, Shibutani M, Mitsumori K, Hanazono Y, Ogawa S, Kurokawa M, Chiba S (2009) Dual antitumor mechanisms of Notch signaling inhibitor in a T-cell acute lymphoblastic leukemia xenograft model. Cancer Sci 100(2):2444–2450. doi:10.1111/j.1349-7006.2009.01328.x

    Article  PubMed  CAS  Google Scholar 

  • Mittal S, Subramanyam D, Dey D, Kumar RV, Rangarajan A (2009) Cooperation of Notch and Ras/MAPK signaling pathways in human breast carcinogenesis. Mol Cancer 8:128. doi:10.1186/1476-4598-8-12

    Article  PubMed  Google Scholar 

  • Moellering RE, Cornejo M, Davis TN, Del Bianco C, Aster JC, Blacklow SC, Kung AL, Gilliland DG, Verdine GL, Bradner JE (2009) Direct inhibition of the NOTCH transcription factor complex. Nature 462(7270):182–188. doi:10.1038/nature08543

    Article  PubMed  CAS  Google Scholar 

  • Nicolas M, Wolfer A, Raj K, Kummer JA, Mill P, van Noort M, Hui CC, Clevers H, Dotto GP, Radtke F (2003) Notch1 functions as a tumor suppressor in mouse skin. Nat Genet 33(3):416–421. doi:10.1038/ng1099

    Article  PubMed  CAS  Google Scholar 

  • Ning L, Wentworth L, Chen H, Weber SM (2009) Down-regulation of Notch1 signaling inhibits tumor growth in human hepatocellular carcinoma. Am J Transl Res 1(4):358–366

    PubMed  CAS  Google Scholar 

  • Pagon Z, Volker J, Cooper GM, Hansen U (2003) Mammalian transcription factor LSF is a target of ERK signaling. J Cell Biochem 89(4):733–746. doi:10.1002/jcb.10549

    Article  PubMed  CAS  Google Scholar 

  • Palomero T, Sulis ML, Cortina M, Real PJ, Barnes K, Ciofani M, Caparros E, Buteau J, Brown K, Perkins SL, Bhagat G, Agarwal AM, Basso G, Castillo M, Nagase S, Cordon-Cardo C, Parsons R, Zúñiga-Pflücker JC, Dominguez M, Ferrando AA (2007) Mutational loss of PTEN induces resistance to NOTCH1 inhibition in T-cell leukemia. Nat Med 13(10):1203–1210. doi:10.1038/nm1636

    Article  PubMed  CAS  Google Scholar 

  • Qi R, An H, Yu Y, Zhang M, Liu S, Xu H, Guo Z, Cheng T, Cao X (2003) Notch1 signaling inhibits growth of human hepatocellular carcinoma through induction of cell cycle arrest and apoptosis. Cancer Res 63(23):8323–8329

    PubMed  CAS  Google Scholar 

  • Rae FK, Stephenson SA, Nicol DL, Clements JA (2000) Novel association of a diverse range of genes with renal cell carcinoma as identified by differential display. Int J Cancer 88(5):726–732. doi:10.1002/1097-0215(20001201)

    Article  PubMed  CAS  Google Scholar 

  • Rao SS, O’Neil J, Liberator CD, Hardwick JS, Dai X, Zhang T, Tyminski E, Yuan J, Kohl NE, Richon VM, Van der Ploeg LH, Carroll PM, Draetta GF, Look AT, Strack PR, Winter CG (2009) Inhibition of NOTCH signaling by gamma secretase inhibitor engages the RB pathway and elicits cell cycle exit in T-cell acute lymphoblastic leukemia cells. Cancer Res 69(7):3060–3068. doi:10.1158/0008-5472

    Article  PubMed  CAS  Google Scholar 

  • Roy M, Pear WS, Aster JC (2007) The multifaceted role of Notch in cancer. Curr Opin Genet Dev 17(1):52–59

    Article  PubMed  CAS  Google Scholar 

  • Saxena UH, Powell CM, Fecko JK, Cacioppo R, Chou HS, Cooper GM, Hansen U (2009) Phosphorylation by cyclin C/cyclin-dependent kinase 2 following mitogenic stimulation of murine fibroblasts inhibits transcriptional activity of LSF during G1 progression. Mol Cell Biol 29(9):2335–2345

    Article  PubMed  CAS  Google Scholar 

  • Shelly LL, Fuchs C, Miele L (1999) Notch-1 prevents apoptosis in murine erythroleukemia cells and is necessary for differentiation induced by hybrid polar drugs. J Cell Biochem 73(2):164–175. doi:10.1002/(SICI)1097-4644(19990501)

    Article  PubMed  CAS  Google Scholar 

  • Shi Y, Chen L, Li J, Lv YL, Sun Q, Wang LX, Jiao SC (2011) Prognostic and predictive values of pERK1/2 and pAkt-1 expression in non-small cell lung cancer patients treated with adjuvant chemotherapy. Tumour Biol 32(2):381–390. doi:10.1007/s13277-010-0131-8

    Article  PubMed  CAS  Google Scholar 

  • Thomas MB, Zhu A (2005) Hepatocellular carcinoma: the need for progress. J Clin Oncol 23(13):2892–2899. doi:10.1200/JCO.2005.03.196

    Article  PubMed  Google Scholar 

  • Tohda S, Nara N (2001) Expression of Notch1 and Jagged1 proteins in acute myeloid leukemia cells. Leuk Lymphoma 42(3):467–472

    Article  PubMed  CAS  Google Scholar 

  • Veljkovic J, Hansen U (2004) Lineage-specific and ubiquitous biological roles of the mammalian transcription factor LSF. Gene 343(1):23–40. doi:10.1016/j.gene.2004.08.010

    Article  PubMed  CAS  Google Scholar 

  • Vilimas T, Mascarenhas J, Palomero T, Mandal M, Buonamici S, Meng F, Thompson B, Spaulding C, Macaroun S, Alegre ML, Kee BL, Ferrando A, Miele L, Aifantis I (2007) Targeting the NF-kB signaling pathway in Notch1-induced T-cell leukemia. Nat Med 13(1):70–77. doi:10.1038/nm1524

    Article  PubMed  CAS  Google Scholar 

  • Volker JL, Rameh LE, Zhu Q, DeCaprio J, Hansen U (1997) Mitogenic stimulation of resting T cells causes rapid phosphorylation of the transcription factor LSF and increased DNA-binding activity. Genes Dev 11(11):1435–1446

    Article  PubMed  CAS  Google Scholar 

  • Wang Z, Li Y, Ahmad A, Banerjee S, Azmi AS, Kong D, Wojewoda C, Miele L, Sarkar FH (2011) Down-regulation of Notch-1 is associated with Akt and FoxM1 in inducing cell growth inhibition and apoptosis in prostate cancer cells. J Cell Biochem 112(1):78–88. doi:10.1002/jcb.22770

    Article  PubMed  CAS  Google Scholar 

  • Weijzen S, Rizzo P, Braid M, Vaishnav R, Jonkheer SM, Zlobin A, Osborne BA, Gottipati S, Aster JC, Hahn WC, Rudolf M, Siziopikou K, Kast WM, Miele L (2002) Activation of Notch-1 signaling maintains the neoplastic phenotype in human Ras-transformed cells. Nat Med 8(9):979–987. doi:10.1038/nm754

    Article  PubMed  CAS  Google Scholar 

  • Weng AP, Nam Y, Wolfe MS, Pear WS, Griffin JD, Blacklow SC, Aster JC (2003) Growth suppression of Pre-T acute lymphoblastic leukemia cells by inhibition of notch signaling. Mol Cell Biol 23(2):655–664. doi:10.1128/MCB.23.2.655-664.2003

    Article  PubMed  CAS  Google Scholar 

  • Westhoff B, Colaluca IN, D’Ario G, Donzelli M, Tosoni D, Volorio S, Pelosi G, Spaggiari L, Mazzarol G, Viale G, Pece S, Di Fiore PP (2009) Alterations of the Notch pathway in lung cancer. Proc Natl Acad Sci 106(52):22293–22298. doi:10.1073/pnas.0907781106

    Article  PubMed  CAS  Google Scholar 

  • Wu Y, Cain-Hom C, Choy L, Hagenbeek TJ, de Leon GP, Chen Y, Finkle D, Venook R, Wu X, Ridgway J, Schahin-Reed D, Dow GJ, Shelton A, Stawicki S, Watts RJ, Zhang J, Choy R, Howard P, Kadyk L, Yan M, Zha J, Callahan CA, Hymowitz SG, Siebel CW (2010) Therapeutic antibody targeting of individual Notch receptors. Nature 464(7291):1052–1059. doi:10.1038/nature08878

    Article  PubMed  CAS  Google Scholar 

  • Zagouras P, Stifani S, Blaumueller CM, Carcangiu ML, Artavanis-Tsakonas S (1995) Alterations in Notch signaling in neoplastic lesions of the human cervix. Proc Natl Acad Sci 92(14):6414–6418

    Article  PubMed  CAS  Google Scholar 

  • Zhang Y, Li B, Ji ZZ, Zheng PS (2010) Notch1 regulates the growth of human colon cancers. Cancer 116(22):5207–5218. doi:10.1002/cncr.25449

    Article  PubMed  CAS  Google Scholar 

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Acknowledgments

We gratefully acknowledge the contributions of Dr.Youwei Zhang and Xiaodong Bu for HCC specimens collection, helpful suggestions and discussions and Dongfeng Liu for paraffin sections manufacture. We thank Dr. Xianfeng Cheng for support on the research. This study is supported by the National Natural Science Foundation of China (No. 30470780).

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We declare that we have no conflict of interest.

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Authors and Affiliations

  1. Department of Pathology, School of Medicine, Southeast University, Nanjing, 210009, China

    Renhua Fan, Pingsheng Chen, Jing Li & Feng Liu

  2. Department of Radiology and nuclear medicine, School of Medicine, Southeast Universtiy, Nanjing, 210009, China

    Di Zhao & Jing long Tong

  3. Department of Clinical Laboratory of Changsha Central Hospital, Changsha, 410004, China

    Renhua Fan

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  1. Renhua Fan
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Correspondence to Pingsheng Chen.

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Fan, R., Chen, P., Zhao, D. et al. Cooperation of deregulated Notch signaling and Ras pathway in human hepatocarcinogenesis. J Mol Hist 42, 473–481 (2011). https://doi.org/10.1007/s10735-011-9353-3

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