Skip to main content

Advertisement

SpringerLink
Log in

Cytokine expression profile in human pancreatic carcinoma cells and in surgical specimens: implications for survival

  • Original Article
  • Published:
Cancer Immunology, Immunotherapy Aims and scope Submit manuscript

Abstract

Cytokine shedding by tumor cells into the local microenvironment modulates host immune response, tumor growth, and metastasis. The study aimed to verify the hypothesis that the immunological microenvironment of pancreatic carcinoma exists in a prevalently immunosuppressive state, influencing survival. We analyzed expression profiles of pro-inflammatory (IL-1β, IL-2, IL-6, IL-8, IL-12 p40, IL-18 and IFN-γ) and anti-inflammatory (IL-10, IL-11, IL-13 and TGF-β isoforms) cytokines. The study was performed both in vitro, in five pancreatic carcinoma cell lines (real time RT-PCR), and in specimens from 65 patients, comparing tumoral versus non-tumoral pancreatic tissues (real time RT-PCR and immunohistochemistry). Furthermore, cytokines were measured in supernatants and sera (from patients and controls) by ELISA. All cell lines expressed IL-8, IL-18, TGF-β1, TGF-β2 and TGF-β3, but not IFN-γ and IL-2 transcripts. Expression of IL-1β, IL-6, IL-10, IL-11, IL-13 and IL-12 mRNA was variable. All the above cytokines were detected as soluble proteins in supernatants, except IL-13. Tumor tissues overexpressed IL-1β, IL-6, IL-8, IL-10, IL-11, IL-12 p40, IL-18, IFN-γ, TGF-β1, TGF-β2 and TGF-β3 at the mRNA level and IL-1β, IL-18, TGF-β2 and TGF-β3 also at the protein level. Conversely, non-tumor tissues had stronger RNA and protein expression of IL-13. Survival was significantly longer in patients with high IL-1β and IL-11 and moderate IL-12 expression. Serum IL-8, IL-10, IL-12, IL-18, TGF-β1 and TGF-β2 were higher in patients than in controls, as opposed to IL-1β and IL-13. Patients with low circulating levels of IL-6, IL-18 and TGF-β2 survived longer. Pancreatic cancer is characterized by peculiar cytokine expression patterns, associated with different survival probabilities.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. Alexandrakis MG, Passam FH, Pappa CA, Sfiridaki K, Tsirakis G, Damilakis J, Stathopoulos EN, Kyriakou DS (2004) Interleukin-18 in multiple myeloma patients: serum levels in relation to response to treatment and survival. Leuk Res 28:259–266

    Article  PubMed  CAS  Google Scholar 

  2. Apte RN, Voronov E (2002) Interleukin-1–a major pleiotropic cytokine in tumor-host interactions. Semin Cancer Biol 12:277–290

    Article  PubMed  CAS  Google Scholar 

  3. Barton BE (2001) IL-6-like cytokines and cancer cachexia: consequences of chronic inflammation. Immunol Res 23:41–58

    Article  PubMed  CAS  Google Scholar 

  4. Bellone G, Smirne C, Carbone A, Mareschi K, Dughera L, Farina EC, Alabiso O, Valente G, Emanuelli G, Rodeck U (2000) Production and pro-apoptotic activity of soluble CD95 ligand in pancreatic carcinoma. Clin Cancer Res 6:2448–2455

    PubMed  CAS  Google Scholar 

  5. Bellone G, Turletti A, Artusio E, Mareschi K, Carbone A, Tibaudi D, Robecchi A, Emanuelli G, Rodeck U (1999) Tumor-associated transforming growth factor-beta and interleukin-10 contribute to a systemic Th2 immune phenotype in pancreatic carcinoma patients. Am J Pathol 155:537–547

    PubMed  CAS  Google Scholar 

  6. Berek JS, Chung C, Kaldi K, Watson JM, Knox RM, Martinez-Maza O (1991) Serum interleukin-6 levels correlate with disease status in patients with epithelial ovarian cancer. Am J Obstet Gynecol 164:1038–1042

    PubMed  CAS  Google Scholar 

  7. Carbone A, Rodeck U, Mauri FA, Sozzi M, Gaspari F, Smirne C, Prati A, Addeo A, Novarino A, Robecchi A, Berretto O, Emanuelli E, Bellone G (2005) Human pancreatic carcinoma cells secrete bioactive Interleukin-18 after treatment with 5-Fluorouracil: Implications for anti-tumor immune response. Cancer Biol Ther 4:231–241

    Article  PubMed  CAS  Google Scholar 

  8. Cuny E, Loiseau H, Penchet G, Ellie E, Arsaut J, Vital A, Vincendeau P, Demotes-Mainard J (2002) Association of elevated glial expression of interleukin-1beta with improved survival in patients with glioblastomas multiforme. J Neurosurg 96:294–301

    PubMed  Google Scholar 

  9. Dinarello CA (1998) Interleukin-1, interleukin-1 receptors and interleukin-1 receptor antagonist. Int Rev Immunol 16:457–499

    Article  PubMed  CAS  Google Scholar 

  10. Fantuzzi G, Dinarello CA (1999) Interleukin-18 and interleukin-1 beta: two cytokine substrates for ICE (caspase-1). J Clin Immunol 19:1–11

    Article  PubMed  CAS  Google Scholar 

  11. Foppoli M, Gianotti L, Galli L, Citterio G, Consogno G, Gentilini O, Braga M (1996) Increased interleukin-10 serum levels in patients with solid tumors. Cancer Lett 104:1–5

    Google Scholar 

  12. Ghayur T, Banerjee S, Hugunin M, Butler D, Herzog L, Carter A, Quintal L, Sekut L, Talanian R, Paskind M, Wong W, Kamen R, Tracey D, Allen H (1997) Caspase-1 processes IFN–inducing factor and regulates LPS-induced IFN- production. Nature 386:619–623

    Article  PubMed  CAS  Google Scholar 

  13. Goggins M, Shekher M, Turnacioglu K, Yeo CJ, Hruban RH, Kern SE (1998) Genetic alterations of the transforming growth factor beta receptor genes in pancreatic and biliary adenocarcinomas. Cancer Res 58:5329–5332

    PubMed  CAS  Google Scholar 

  14. Huang M, Wang J, Lee P, Sharma S, Mao JT, Meissner H, Uyemura K, Modlin R, Wollman J, Dubinett SM (1995) Human non-small cell lung cancer cells express a type 2 cytokine pattern. Cancer Res 55:3847–3853

    PubMed  CAS  Google Scholar 

  15. Kawabata T, Ichikura T, Majima T, Seki S, Chochi K, Takayama E, Hiraide H, Mochizuki H (2001) Preoperative serum interleukin-18 levels as a postoperative prognostic marker in patients with gastric carcinoma. Cancer 92:2050–2055

    Article  PubMed  CAS  Google Scholar 

  16. Kawakami Y, Okada T, Akada M (2004) Development of immunotherapy for pancreatic cancer. Pancreas 28:320–325

    Article  PubMed  CAS  Google Scholar 

  17. Kleeff J, Ishiwata T, Maruyama H, Friess H, Truong P, Buchler MW, Falb D, Korc M (1999) The TGF-beta signaling inhibitor Smad7 enhances tumorigenicity in pancreatic cancer. Oncogene 18:5363–5372

    Article  PubMed  CAS  Google Scholar 

  18. Kruger-Krasagakes S, Krasagakis K, Garbe C, Schmitt E, Huls C, Blankenstein T, Diamantstein T (1994) Expression of interleukin 10 in human melanoma. Br J Cancer 70:1182–1185

    PubMed  CAS  Google Scholar 

  19. Landis SH, Murray T, Bolden S, Wingo PA (1999) Cancer statistics. CA Cancer J Clin 49:8–31

    PubMed  CAS  Google Scholar 

  20. Lattime EC, McCue PA, Keely FX, Li W, Gomella LG (1995) Expression of IL-10 mRNA in biopsies of superficial and invasive TCC of the human bladder. Proc Am Assoc Cancer Res 36:462

    Google Scholar 

  21. Lawrence DA (1996) Transforming growth factor-beta: a general review. Eur Cytokine Netw 7:363–374

    PubMed  CAS  Google Scholar 

  22. Lebel-Binay S, Thiounn N, De Pinieux G, Vieillefond A, Debre B, Bonnefoy JY, Fridman WH, Pages F (2003) IL-18 is produced by prostate cancer cells and secreted in response to interferons. Int J Cancer 106:827–835

    Article  PubMed  CAS  Google Scholar 

  23. Li A, Dubey S, Varney ML, Dave BJ, Singh RK (2003) IL-8 directly enhanced endothelial cell survival, proliferation, and matrix metalloproteinases production and regulated angiogenesis. J Immunol 170:3369–3376

    PubMed  CAS  Google Scholar 

  24. Ling P, Gately MK, Gubler U, Stern AS, Lin P, Hollfelder K, Su C, Pan YC, Hakimi J (1995) Human IL-12 p40 homodimer binds to the IL-12 receptor but does not mediate biologic activity. J Immunol 154:116–127

    PubMed  CAS  Google Scholar 

  25. Lissoni P, Brivio F, Rovelli F, Fumagalli G, Malugani F, Vaghi M, Secondino S, Bucovec R, Gardani GS (2000) Serum concentrations of interleukin-18 in early and advanced cancer patients: enhanced secretion in metastatic disease. J Biol Regul Homeost Agents 14:275–277

    PubMed  CAS  Google Scholar 

  26. Lu C, Kerbel RS (1993) Interleukin-6 undergoes transition from paracrine growth inhibitor to autocrine stimulator during human melanoma progression. J Cell Biol 120:1281–1288

    Article  PubMed  CAS  Google Scholar 

  27. MacKenzie MJ (2004) Molecular therapy in pancreatic adenocarcinoma. Lancet Oncol 5:541–549

    Article  PubMed  CAS  Google Scholar 

  28. Maeda H, Shiraishi A (1996) TGF-beta contributes to the shift toward Th2-type responses through direct and IL-10-mediated pathways in tumor-bearing mice. J Immunol 156:73–78

    PubMed  CAS  Google Scholar 

  29. Martone T, Bellone G, Pagano M, Beatrice F, Palonta F, Emanuelli G, Cortesina G (2004) Constitutive expression of interleukin-18 in head and neck squamous carcinoma cells. Head & Neck 26:494–503

    Article  PubMed  Google Scholar 

  30. Mattner F, Fischer S, Guckes S, Jin S, Kaulen H, Schmitt E, Rude E, Germann T (1993) The interleukin-12 subunit p40 specifically inhibits effects of the interleukin-12 heterodimer. Eur J Immunol 23:2202–2208

    Article  PubMed  CAS  Google Scholar 

  31. Michalaki V, Syrigos K, Charles P, Waxman J (2004) Serum levels of IL-6 and TNF-alpha correlate with clinicopathological features and patient survival in patients with prostate cancer. Br J Cancer 90:2312–2316

    PubMed  CAS  Google Scholar 

  32. Moore MB, Kurago ZB, Fullenkamp CA, Lutz CT (2003) Squamous cell carcinoma cells differentially stimulate NK cell effector functions: the role of IL-18. Cancer Immunol Immunother 52:107–115

    PubMed  CAS  Google Scholar 

  33. Nakagomi H, Pisa P, Pisa EK, Yamamoto Y, Halapi E, Backlin K, Juhlin C, Kiessling R (1995) Lack of interleukin-2 (IL-2) expression and selective expression of IL-10 mRNA in human renal cell carcinoma. Int J Cancer 63:366–371

    Article  PubMed  CAS  Google Scholar 

  34. Nakanishi K, Yoshimoto T, Tsutsui H, Okamura H (2001) Interleukin-18 is a unique cytokine that stimulates both Th1 and Th2 responses depending on its cytokine milieu. Cytokine Growth Factor Rev 12:53–72

    Article  PubMed  CAS  Google Scholar 

  35. Okamura H, Tsutsi H, Komatsu T, Yutsudo M, Hakura A, Tanimoto T, Torigoe K, Okura T, Nukada Y, Hattori K, Akita K, Namba M, Tanabe F, Konishi K, Fukuda S, Kurimoto M (1995) Cloning of a new cytokine that induces IFN- production by T cells. Nature 378:88–91

    Article  PubMed  CAS  Google Scholar 

  36. Pages F, Berger A, Henglein B, Piqueras B, Danel C, Zinzindohoue F, Thiounn N, Cugnenc PH, Fridman WH (1999) Modulation of interleukin-18 expression in human colon carcinoma: consequences for tumor immune surveillance. Int J Cancer 84:326–330

    Article  PubMed  CAS  Google Scholar 

  37. Paglia D, Oran A, Lu C, Kerbel RS, Sauder DN, McKenzie RC (1995) Expression of leukemia inhibitory factor and interleukin-11 by human melanoma cell lines: LIF, IL-6, and IL-11 are not coregulated. J Interferon Cytokine Res 15:455–460

    PubMed  CAS  Google Scholar 

  38. Park H, Byun D, Kim TS, Kim YI, Kang JS, Hahm ES, Kim SH, Lee WJ, Song HK, Yoon DY, Kang CJ, Lee C, Houh D, Kim H, Cho B, Kim Y, Yang YH, Min KH, Cho DH (2001) Enhanced IL-18 expression in common skin tumors. Immunol Lett 79:215–219

    Article  PubMed  CAS  Google Scholar 

  39. Pfaffl MW, Horgan GW, Dempfle L (2002) Relative expression software tool (REST) for group-wise comparison and statistical analysis of relative expression results in real-time PCR. Nucleic Acids Res 30:e36

    Article  PubMed  Google Scholar 

  40. Ramakers C, Ruijter JM, Deprez RH, Moorman AF (2003) Assumption-free analysis of quantitative real-time polymerase chain reaction (PCR) data. Neurosci Lett 339:62–66

    Article  PubMed  CAS  Google Scholar 

  41. Rasmussen R (2001) Quantification on the LightCycler. In: Meuer S, Wittwer C, Nakagawara K (eds) Rapid Cycle Real-time PCR, Methods and Applications. Springer Press, Heidelberg, pp 21–34

    Google Scholar 

  42. Riedel F, Adam S, Feick P, Haas S, Gotte K, Hormann K (2004) Mannheim Alcohol Study Group. Expression of IL-18 in patients with head and neck squamous cell carcinoma. Int J Mol Med 13:267–272

    Google Scholar 

  43. Saito H, Tsujitani S, Oka S, Kondo A, Ikeguchi M, Maeta M, Kaibara N (2000) An elevated serum level of transforming growth factor-beta 1 (TGF-beta 1) significantly correlated with lymph node metastasis and poor prognosis in patients with gastric carcinoma. Anticancer Res 20:4489–4493

    PubMed  CAS  Google Scholar 

  44. Sato T, McCue P, Masuoka K, Salwen S, Lattime EC, Mastrangelo MJ, Berd D (1996) Interleukin 10 production by human melanoma. Clin Cancer Res 2:1383–1390

    PubMed  CAS  Google Scholar 

  45. Scupoli MT, Sartoris S, Tosi G, Ennas MG, Nicolis M, Cestari T, Zamboni G, Martignoni G, Lemoine NR, Scarpa A, Accolla RS (1996) Expression of MHC class I and class II antigens in pancreatic adenocarcinomas. Tissue Antigens 48:301–311

    Article  PubMed  CAS  Google Scholar 

  46. Seddighzadeh M, Larsson P, Ulfgren AC, Onelov E, Berggren P, Tribukait B, Torstensson A, Norming U, Wijkstrom H, Linder S, Steineck G (2003) Low IL-1alpha expression in bladder cancer tissue and survival. Eur Urol 43:362–368

    Article  PubMed  CAS  Google Scholar 

  47. Sobin LH, Wittekind CL (2002) (eds) International Union Against Cancer (UICC): TNM classification of malignant tumors. John Wiley & Sons Inc New York, pp. 93–96

  48. Sotiriou C, Lacroix M, Lespagnard L, Larsimont D, Paesmans M, Body JJ (2001) Interleukins-6 and −11 expression in primary breast cancer and subsequent development of bone metastases. Cancer Lett 169:87–95

    Article  PubMed  CAS  Google Scholar 

  49. Sozen S, Coskun U, Sancak B, Bukan N, Gunel N, Tunc L, Bozkirli I (2004) Serum levels of interleukin-18 and nitrite+nitrate in renal cell carcinoma patients with different tumor stage and grade. Neoplasma 51:25–29

    PubMed  CAS  Google Scholar 

  50. Terabe M, Park JM, Berzofsky JA (2004) Role of IL-13 in regulation of anti-tumor immunity and tumor growth. Cancer Immunol Immunother 53:79–85

    Article  PubMed  CAS  Google Scholar 

  51. Tohyama K, Yoshida Y, Ohashi K, Sano E, Kobayashi H, Endo K, Naruto M, Nakamura T (1992) Production of multiple growth factors by a newly established human thyroid carcinoma cell line. Jpn J Cancer Res 83:153–158

    PubMed  CAS  Google Scholar 

  52. Torroella-Kouri M, Keith JC, Ivanova M, Lopez DM (2003) IL-11-induced reduction of C/EBP transcription factor binding may contribute to the IL-12 downregulation in tumor-bearing mice. Int J Oncol 22:439–448

    PubMed  CAS  Google Scholar 

  53. Trepicchio WL, Bozza M, Pedneault G, Dorner AJ (1996) Recombinant human IL-11 attenuates the inflammatory response through down-regulation of proinflammatory cytokine release and nitric oxide production. J Immunol 157:3627–3634

    PubMed  CAS  Google Scholar 

  54. Trinchieri G (2003) Interleukin-12 and the regulation of innate resistance and adaptive immunity. Nat Rev Immunol 3:133–146

    Article  PubMed  CAS  Google Scholar 

  55. Tsuboi K, Miyazaki T, Nakajima M, Fukai Y, Masuda N, Manda R, Fukuchi M, Kato H, Kuwano H (2004) Serum interleukin-12 and interleukin-18 levels as a tumor marker in patients with esophageal carcinoma. Cancer Lett 205:207–214

    Article  PubMed  CAS  Google Scholar 

  56. Ueda T, Shimada E, Urakawa T (1994) Serum levels of cytokines in patients with colorectal cancer: possible involvement of interleukin-6 and interleukin-8 in hematogenous metastasis. J Gastroenterol 29:423–429

    Article  PubMed  CAS  Google Scholar 

  57. Ungefroren H, Voss M, Jansen M, Roeder C, Henne-Bruns D, Kremer B, Kalthoff H (1998) Human pancreatic adenocarcinomas express Fas and Fas ligand yet are resistant to Fas-mediated apoptosis. Cancer Res 58: 1741–1749

    PubMed  CAS  Google Scholar 

  58. Ungefroren H, Voss M, Bernstorff WV, Schmid A, Kremer B, Kalthoff H (1999) Immunological escape mechanisms in pancreatic carcinoma. Ann N Y Acad Sci 880:243–251

    Article  PubMed  CAS  Google Scholar 

  59. von Bernstorff W, Spanjaard RA, Chan AK, Lockhart DC, Sadanaga N, Wood I, Peiper M, Goedegebuure PS, Eberlein TJ (1999) Pancreatic cancer cells can evade immune surveillance via nonfunctional Fas (APO-1/CD95) receptors and aberrant expression of functional Fas ligand. Surgery 125: 73–84

    Article  Google Scholar 

  60. von Bernstorff W, Voss M, Freichel S, Schmid A, Vogel I, Johnk C, Henne-Bruns D, Kremer B, Kalthoff H (2001) Systemic and local immunosuppression in pancreatic cancer patients. Clin Cancer Res 7(Suppl): 925s-932s

    Google Scholar 

  61. Wang ZY, Gaggero A, Rubartelli A, Rosso O, Miotti S, Mezzanzanica D, Canevari S, Ferrini S (2002) Expression of interleukin-18 in human ovarian carcinoma and normal ovarian epithelium: evidence for defective processing in tumor cells. Int J Cancer 98:873–878

    Article  PubMed  CAS  Google Scholar 

  62. Wakeman CJ, Martin IG, Robertson RW, Dobbs BR, Frizelle FA (2004) Pancreatic cancer: management and survival. ANZ J Surg 74:941–944

    Article  PubMed  Google Scholar 

  63. Xu J, Menezes J, Prasad U, Ahmad A (1999) Elevated serum levels of transforming growth factor beta1 in Epstein-Barr virus-associated nasopharyngeal carcinoma patients. Int J Cancer 84:396–399

    Article  PubMed  CAS  Google Scholar 

  64. Zhang B, Ma XT, Zheng GG, Li G, Rao Q, Wu KF (2003) Expression of IL-18 and its receptor in human leukemia cells. Leuk Res 27:813–822

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

This work was supported by a grant from MIUR (Rome, Italy) (ex-60%) to GE, and in part by a grant from the Piedmontese Regional Government (Regione Piemonte) to GB. ET and AB are the recipients of an award from the Piedmontese Regional Government.

Author information

Authors and Affiliations

  1. Department of Clinical Physiopathology, Università di Torino, Via Genova, 3, 10126, Torino, Italy

    Graziella Bellone, Carlo Smirne, Elena Tonel, Anna Carbone, Alessandra Buffolino & Giorgio Emanuelli

  2. Department of Medical-Surgical Disciplines, University of Turin, Turin, Italy

    Antonio Robecchi

  3. Medical Sciences, University of Eastern Piedmont, Novara, Italy

    Carlo Smirne & Mario Pirisi

  4. Gastroenterology and Clinical Nutrition, S. Giovanni Battista Hospital, Turin, Italy

    Anna Carbone, Luca Dughera & Giorgio Emanuelli

  5. Histopathology, Hammersmith Hospital, London, UK

    Francesco Angelo Mauri

Authors
  1. Graziella Bellone
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Carlo Smirne
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Francesco Angelo Mauri
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Elena Tonel
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Anna Carbone
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Alessandra Buffolino
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Luca Dughera
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Antonio Robecchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Mario Pirisi
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Giorgio Emanuelli
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Graziella Bellone.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Bellone, G., Smirne, C., Mauri, F.A. et al. Cytokine expression profile in human pancreatic carcinoma cells and in surgical specimens: implications for survival. Cancer Immunol Immunother 55, 684–698 (2006). https://doi.org/10.1007/s00262-005-0047-0

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00262-005-0047-0

Keywords

Search

Navigation