Targeted Oncology

, Volume 13, Issue 5, pp 657–665 | Cite as

Vitamin D Deficiency has a Negative Impact on Cetuximab-Mediated Cellular Cytotoxicity against Human Colon Carcinoma Cells

  • Lorenzo Mortara
  • Marzia B. Gariboldi
  • Annalisa Bosi
  • Marco Bregni
  • Graziella Pinotti
  • Luigina Guasti
  • Alessandro Squizzato
  • Douglas M. Noonan
  • Elena MontiEmail author
  • Leonardo Campiotti
Original Research Article



Hypovitaminosis D is associated with an adverse prognosis in colon cancer patients, possibly due to the effects of the vitamin on the immune system. Antibody-dependent cell-mediated cytotoxicity (ADCC) significantly contributes to the anti-tumor effects of monoclonal antibodies, including cetuximab, an epidermal growth factor receptor (EGFR)-targeted monoclonal antibody that is frequently added to chemotherapy in the treatment of colon cancer.


The present study evaluates the association between vitamin D serum levels and the ability of ex vivo NK cells to support cetuximab-mediated ADCC in colon cancer cell lines.


Blood samples were obtained from 124 healthy volunteers and serum vitamin D was determined by RIA. NK cells were isolated from each sample and added to human colorectal carcinoma cells with or without cetuximab, and ADCC was assessed using a colorimetric lactate dehydrogenase assay.


Correlation analysis indicates a significant, gender- and age-independent association between vitamin D levels and cetuximab-induced ADCC on HT29 cells, where NK cells from samples with vitamin D < 20 ng/mL are significantly less efficient in inducing ADCC. A confirmatory study on two additional colon cancer cell lines yielded similar results.


These data suggest that vitamin D supplementation in vitamin-deficient/insufficient colorectal cancer patients could improve cetuximab-induced ADCC.



The Authors thank Ambra Fortunato, Marì De Leo and Emanuela Marras for technical support.

Compliance with Ethical Standards


This study was supported by the Università degli Studi dell’Insubria, Fondi di Ateneo per la Ricerca “FAR 2014” and “FAR2015” (to LM) and the Italian Association for Cancer Research (AIRC- IG15895 to DN). The funding agencies were not involved in the research plan and the interpretation of the data.

Conflict of Interest

Lorenzo Mortara, Marzia Bruna Gariboldi, Annalisa Bosi, Marco Bregni, Graziella Pinotti, Luigina Guasti, Alessandro Squizzato, Douglas Noonan, Elena Monti, and Leonardo Campiotti declare that they have no conflicts of interest that might be relevant to the contents of this manuscript.


  1. 1.
    Kennel KA, Drake MT, Hurley DL. Vitamin D deficiency in adults: when to test and how to treat. Mayo Clin Proc. 2010;85(8):752–7.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Christakos S, Dhawan P, Verstuyf A, Verlinden L, Carmeliet G. Vitamin D: metabolism, molecular mechanism of action, and pleiotropic effects. Physiol Rev. 2016;96(1):365–408.CrossRefPubMedGoogle Scholar
  3. 3.
    Morán-Auth Y, Penna-Martinez M, Shoghi F, Ramos-Lopez E, Badenhoop K. Vitamin D status and gene transcription in immune cells. J Steroid Biochem Mol Biol. 2013;136:83–5.CrossRefPubMedGoogle Scholar
  4. 4.
    Verstuyf A, Carmeliet G, Bouillon R, Mathieu C. Vitamin D: a pleiotropic hormone. Kidney Int. 2010;78(2):140–5.CrossRefPubMedGoogle Scholar
  5. 5.
    Holick MF. Vitamin D deficiency. N Engl J Med. 2007;357(3):266–81.CrossRefPubMedGoogle Scholar
  6. 6.
    Kennel KA, Drake MT. Vitamin D in the cancer patient. Curr Opin Support Palliat Care. 2013;7(3):272–7.PubMedPubMedCentralGoogle Scholar
  7. 7.
    Fang S, Sui D, Wang Y, Liu H, Chiang YJ, Ross MI, et al. Association of vitamin D levels with outcome in patients with melanoma after adjustment for C-reactive protein. J Clin Oncol. 2016;34(15):1741–7.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Wang W, Li G, He X, Gao J, Wang R, Wang Y, et al. Serum 25-hydroxyvitamin D levels and prognosis in hematological malignancies: a systematic review and meta-analysis. Cell Physiol Biochem. 2015;35(5):1999–2005.CrossRefPubMedGoogle Scholar
  9. 9.
    Li M, Chen P, Li J, Chu R, Xie D, Wang H. Review: the impacts of circulating 25-hydroxyvitamin D levels on cancer patient outcomes: a systematic review and meta-analysis. J Clin Endocrinol Metab. 2014;99(7):2327–36.CrossRefPubMedGoogle Scholar
  10. 10.
    Yao S, Kwan ML, Ergas IJ, Roh JM, Cheng TD, Hong CC, et al. Association of Serum Level of vitamin D at diagnosis with breast Cancer survival: a case-cohort analysis in the pathways study. JAMA Oncol. 2017;3(3):351–7.CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Fuchs MA, Yuan C, Sato K, Niedzwiecki D, Ye X, Saltz LB, et al. Predicted vitamin D status and colon cancer recurrence and mortality in CALGB 89803 (alliance). Ann Oncol. 2017;28(6):1359–67.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Kelly JL, Salles G, Goldman B, Fisher RI, Brice P, Press O, et al. Low serum vitamin D levels are associated with inferior survival in follicular lymphoma: a prospective evaluation in SWOG and LYSA studies. J Clin Oncol. 2015;33(13):1482–90.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Van Loon K, Owzar K, Jiang C, Kindler HL, Mulcahy MF, Niedzwiecki D, et al. 25-Hydroxyvitamin D levels and survival in advanced pancreatic cancer: findings from CALGB 80303 (Alliance). J Natl Cancer Inst. 2014;106(8). pii: dju185.Google Scholar
  14. 14.
    Bessler H, Djaldetti M. 1α,25-Dihydroxyvitamin D3 modulates the interaction between immune and colon cancer cells. Biomed Pharmacother. 2012;66(6):428–32.CrossRefPubMedGoogle Scholar
  15. 15.
    Siegel RL, Miller KD, Jemal A. Cancer statistics, 2017. CA Cancer J Clin. 2017;67(1):7–30.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    O'Connell JB, Maggard MA, Ko CY. Colon cancer survival rates with the new American joint committee on Cancer sixth edition staging. J Natl Cancer Inst. 2004;96(19):1420–5.CrossRefPubMedGoogle Scholar
  17. 17.
    Vogel A, Hofheinz RD, Kubicka S, Arnold D. Treatment decisions in metastatic colorectal cancer - beyond first and second line combination therapies. Cancer Treat Rev. 2017;59:54–60.CrossRefPubMedGoogle Scholar
  18. 18.
    Ciombor KK, Bekaii-Saab T. A comprehensive review of sequencing and combination strategies of targeted agents in metastatic colorectal cancer. Oncologist. 2018;23(1):25–34.CrossRefPubMedGoogle Scholar
  19. 19.
    Brandi G, De Lorenzo S, Nannini M, Curti S, Ottone M, Dall'Olio FG, et al. Adjuvant chemotherapy for resected colorectal cancer metastases: literature review and meta-analysis. World J Gastroenterol. 2016;22(2):519–33.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Sartore-Bianchi A, Loupakis F, Argiles G, Prager GW. Challenging chemoresistant metastatic colorectal cancer: therapeutic strategies from the clinic and from the laboratory. Ann Oncol. 2016;27(8):1456–66.CrossRefPubMedGoogle Scholar
  21. 21.
    Capdevila J, Elez E, Macarulla T, Ramos FJ, Ruiz-Echarri M, Tabernero J. Anti-epidermal growth factor receptor monoclonal antibodies in cancer treatment. Cancer Treat Rev. 2009;35(4):354–63.CrossRefPubMedGoogle Scholar
  22. 22.
    Fearon ER. Molecular genetics of colorectal cancer. Annu Rev Pathol. 2011;6:479–507.CrossRefPubMedGoogle Scholar
  23. 23.
    Shaib W, Mahajan R, El-Rayes B. Markers of resistance to anti-EGFR therapy in colorectal cancer. J Gastrointest Oncol. 2013;4(3):308–18.PubMedPubMedCentralGoogle Scholar
  24. 24.
    Sforza V, Martinelli E, Ciardiello F, Gambardella V, Napolitano S, Martini G, et al. Mechanisms of resistance to anti-epidermal growth factor receptor inhibitors in metastatic colorectal cancer. World J Gastroenterol. 2016;22(28):6345–61.CrossRefPubMedPubMedCentralGoogle Scholar
  25. 25.
    Monteverde M, Milano G, Strola G, Maffi M, Lattanzio L, Vivenza D, et al. The relevance of ADCC for EGFR targeting: a review of the literature and a clinically-applicable method of assessment in patients. Crit Rev Oncol Hematol. 2015;95(2):179–90.CrossRefPubMedGoogle Scholar
  26. 26.
    Inoue Y, Hazama S, Suzuki N, Tokumitsu Y, Kanekiyo S, Tomochika S, et al. Cetuximab strongly enhances immune cell infiltration into liver metastatic sites in colorectal cancer. Cancer Sci. 2017;108(3):455–60.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Lo Nigro C, Ricci V, Vivenza D, Monteverde M, Strola G, Lucio F, et al. Evaluation of antibody-dependent cell-mediated cytotoxicity activity and cetuximab response in KRAS wild-type metastatic colorectal cancer patients. World J Gastrointest Oncol. 2016;8(2):222–30.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Taylor RJ, Saloura V, Jain A, Goloubeva O, Wong S, Kronsberg S, et al. Ex vivo antibody-dependent cellular cytotoxicity inducibility predicts efficacy of cetuximab. Cancer Immunol Res. 2015;3(5):567–74.CrossRefPubMedPubMedCentralGoogle Scholar
  29. 29.
    Trotta AM, Ottaiano A, Romano C, Nasti G, Nappi A, De Divitiis C, et al. Prospective evaluation of Cetuximab-mediated antibody-dependent cell cytotoxicity in metastatic colorectal Cancer patients predicts treatment efficacy. Cancer Immunol Res. 2016;4(4):366–74.CrossRefPubMedGoogle Scholar
  30. 30.
    Veluchamy JP, Spanholtz J, Tordoir M, Thijssen VL, Heideman DA, Verheul HM, et al. Combination of NK cells and Cetuximab to enhance anti-tumor responses in RAS mutant metastatic colorectal Cancer. PLoS One. 2016;11(6):e0157830.CrossRefPubMedPubMedCentralGoogle Scholar
  31. 31.
    Bittenbring JT, Neumann F, Altmann B, Achenbach M, Reichrath J, Ziepert M, et al. Vitamin D deficiency impairs rituximab-mediated cellular cytotoxicity and outcome of patients with diffuse large B-cell lymphoma treated with but not without rituximab. J Clin Oncol. 2014;32(29):3242–8.CrossRefPubMedGoogle Scholar
  32. 32.
    Ross AC, Manson JE, Abrams SA, Aloia JF, Brannon PM, Clinton SK, et al. The 2011 report on dietary reference intakes for calcium and vitamin D from the Institute of Medicine: what clinicians need to know. J Clin Endocrinol Metab. 2011;96(1):53–8.CrossRefPubMedGoogle Scholar
  33. 33.
    Cashman KD, Dowling KG, Škrabáková Z, Gonzalez-Gross M, Valtueña J, De Henauw S, et al. Vitamin D deficiency in Europe: pandemic? Am J Clin Nutr. 2016;103(4):1033–44.CrossRefPubMedPubMedCentralGoogle Scholar
  34. 34.
    Rosen CJ. Clinical practice. Vitamin D insufficiency. N Engl J Med. 2011;364(3):248–54.CrossRefPubMedGoogle Scholar
  35. 35.
    Teleni L, Baker J, Koczwara B, Kimlin MG, Walpole E, Tsai K, et al. Clinical outcomes of vitamin D deficiency and supplementation in cancer patients. Nutr Rev. 2013;71(9):611–21.CrossRefPubMedGoogle Scholar
  36. 36.
    DeMille DM, Piscitelli M, Ocker A, Vuong C, Hartner L, Lynch MP. Vitamin D deficiency in the oncology setting. J Community Support Oncol. 2014;12(1):13–9.CrossRefPubMedGoogle Scholar
  37. 37.
    Ahmad II, Trikudanathan G, Feinn R, Anderson JC, Nicholson M, Lowe S, et al. Low serum vitamin D: a surrogate marker for advanced Colon adenoma? J Clin Gastroenterol. 2016;50(8):644–8.CrossRefPubMedGoogle Scholar
  38. 38.
    Skender S, Bohm J, Schrotz-King P, Chang-Claude J, Siegel EM, Steindorf K, et al. Plasma 25-Hydroxyvitamin D3 levels in colorectal Cancer patients and associations with physical activity. Nutr Cancer. 2017;69(2):229–37.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Maalmi H, Walter V, Jansen L, Chang-Claude J, Owen RW, Ulrich A, et al. Relationship of very low serum 25-hydroxyvitamin D3 levels with long-term survival in a large cohort of colorectal cancer patients from Germany. Eur J Epidemiol. 2017;32(11):961–971.CrossRefPubMedGoogle Scholar
  40. 40.
    Maalmi H, Ordóñez-Mena JM, Schöttker B, Brenner H. Serum 25-hydroxyvitamin D levels and survival in colorectal and breast cancer patients: systematic review and meta-analysis of prospective cohort studies. Eur J Cancer. 2014;50(8):1510–21.CrossRefPubMedGoogle Scholar
  41. 41.
    McCullough ML, Zoltick ES, Weinstein SJ, Fedirko V, Wang M, Cook NR, et al. Circulating vitamin D and colorectal cancer risk: an international pooling project of 17 cohorts. J Natl Cancer Inst. 2018.
  42. 42.
    Colotta F, Jansson B, Bonelli F. Modulation of inflammatory and immune responses by vitamin D. J Autoimmun. 2017;85:78–97.CrossRefPubMedGoogle Scholar
  43. 43.
    Kimmie N, VA P, Kaori S, Chen Y, HB W, Donna N, et al. Vitamin D status and survival of metastatic colorectal cancer patients: results from CALGB/SWOG 80405 (Alliance). J Clin Oncol. 2015;33(15 S)3503–3503.Google Scholar
  44. 44.
    Weeres MA, Robien K, Ahn YO, Neulen ML, Bergerson R, Miller JS, et al. The effects of 1,25-dihydroxyvitamin D3 on in vitro human NK cell development from hematopoietic stem cells. J Immunol. 2014;193(7):3456–62.CrossRefPubMedPubMedCentralGoogle Scholar
  45. 45.
    Merino F, Alvarez-Mon M, de la Hera A, Alés JE, Bonilla F, Durantez A. Regulation of natural killer cytotoxicity by 1,25-dihydroxyvitamin D3. Cell Immunol. 1989;118(2):328–36.CrossRefPubMedGoogle Scholar
  46. 46.
    Buchau AS, Morizane S, Trowbridge J, Schauber J, Kotol P, Bui JD, et al. The host defense peptide cathelicidin is required for NK cell-mediated suppression of tumor growth. J Immunol. 2010;184(1):369–78.CrossRefPubMedGoogle Scholar
  47. 47.
    Lee JH, Park S, Cheon S, Kim S, Hur DY, Kim TS, et al. 1,25-Dihydroxyvitamin D3 enhances NK susceptibility of human melanoma cells via Hsp60-mediated FAS expression. Eur J Immunol. 2011;41(10):2937–46.CrossRefPubMedGoogle Scholar
  48. 48.
    Pander J, Heusinkveld M, van der Straaten T, Jordanova ES, Baak-Pablo R, Gelderblom H, et al. Activation of tumor-promoting type 2 macrophages by EGFR-targeting antibody cetuximab. Clin Cancer Res. 2011;17(17):5668–73.CrossRefPubMedGoogle Scholar
  49. 49.
    Bruns H, Büttner M, Fabri M, Mougiakakos D, Bittenbring JT, Hoffmann MH, et al. Vitamin D-dependent induction of cathelicidin in human macrophages results in cytotoxicity against high-grade B cell lymphoma. Sci Transl Med. 2015;7(282):282ra47.CrossRefPubMedGoogle Scholar

Copyright information

© Springer Nature Switzerland AG 2018

Authors and Affiliations

  • Lorenzo Mortara
    • 1
  • Marzia B. Gariboldi
    • 2
  • Annalisa Bosi
    • 1
  • Marco Bregni
    • 3
  • Graziella Pinotti
    • 4
  • Luigina Guasti
    • 5
  • Alessandro Squizzato
    • 5
  • Douglas M. Noonan
    • 1
    • 6
  • Elena Monti
    • 2
    Email author
  • Leonardo Campiotti
    • 5
  1. 1.Immunology and General Pathology Laboratory, Department of Biotechnology and Life SciencesUniversità degli Studi dell’InsubriaVareseItaly
  2. 2.Laboratory of Anticancer Pharmacology, Department of Biotechnology and Life SciencesUniversità degli Studi dell’InsubriaBusto ArsizioItaly
  3. 3.Medical OncologyASST Valle OlonaVareseItaly
  4. 4.Medical OncologyASST Sette LaghiVareseItaly
  5. 5.Department of Medicine and SurgeryUniversità degli Studi dell’InsubriaVareseItaly
  6. 6.Scientific and Technology PoleIRCCS MultiMedicaMilanItaly

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