Pathology & Oncology Research

, Volume 23, Issue 2, pp 335–343 | Cite as

Alpha-1-Antitrypsin Antagonizes Cisplatin-Induced Cytotoxicity in Prostate Cancer (PC3) and Melanoma Cancer (A375) Cell Lines

  • Mila Ljujic
  • Sanja Mijatovic
  • Mirna Z. Bulatovic
  • Marija Mojic
  • Danijela Maksimovic-Ivanic
  • Dragica Radojkovic
  • Aleksandra TopicEmail author
Original Article


Increased circulating alpha-1-antitrypsin (AAT) correlates with cancer stage/aggressiveness, but its role in cancer biology is unclear. We revealed antagonistic effect of AAT to cisplatin-induced cytotoxicity in prostate (PC3) and melanoma (A375) cancer cell lines. Moreover, AAT abrogated cytotoxicity of MEK inhibitor U0126 in PC3 cell line. Weaker antagonistic effect of AAT on cytotoxicity of PI3/Akt and NF-kB inhibitors was also observed. In addition, cisplatin increased AAT gene expression in transfected PC3 cells. However, AAT derived from transfected PC3 cells did not antagonize cisplatin-induced cytotoxicity. In conclusion, these results suggest possible association between high circulating AAT and cisplatin resistance.


Alpha-1-antitrypsin Cisplatin Prostate cancer Melanoma cancer 



This work was supported by the Serbian Ministry of Education and Science (Grants No 173013 and 173008). Authors wish to thank Dr. Djordje Miljkovic from Institute for Biological Research “Sinisa Stankovic”, University of Belgrade for help in performing flow cytometry and Dr. Stanislava Stosic Grujicic from Institute for Biological Research “Sinisa Stankovic”, University of Belgrade for insightful comments and critical reading of the manuscript.

Compliance with Ethical Standards

Conflict of Interest

The authors have declared no conflicts of interest.


  1. 1.
    Crystal RG (1989) The alpha 1-antitrypsin gene and its deficiency states. Trends Genet 5:411–417CrossRefPubMedGoogle Scholar
  2. 2.
    Blank CA, Brantly M (1994) Clinical features and molecular characteristics of alpha 1-antitrypsin deficiency. Ann Allergy 72:105–120PubMedGoogle Scholar
  3. 3.
    Mason DY, Cramer EM, Massé JM, Crystal R, Bassot JM, Breton-Gorius J (1991) Alpha 1-antitrypsin is present within the primary granules of human polymorphonuclear leukocytes. Am J Pathol 139:623–628PubMedPubMedCentralGoogle Scholar
  4. 4.
    Molmenti EP, Perlmutter DH, Rubin DC (1993) Cell-specific expression of alpha 1-antitrypsin in human intestinal epithelium. J Clin Invest 92:2022–2034CrossRefPubMedPubMedCentralGoogle Scholar
  5. 5.
    Perlmutter DH, Daniels JD, Auerbach HS, De Schryver-Kecskemeti K, Winter HS, Alpers DH (1989) The alpha 1-antitrypsin gene is expressed in a human intestinal epithelial cell line. J Biol Chem 264:9485–9490PubMedGoogle Scholar
  6. 6.
    Sallenave JM, Tremblay GM, Gauldie J, Richards CD (1997) Oncostatin M, but not interleukin-6 or leukemia inhibitory factor, stimulates expression of alpha1-proteinase inhibitor in A549 human alveolar epithelial cells. J Interf Cytokine Res 17:337–346CrossRefGoogle Scholar
  7. 7.
    Cichy J, Potempa J, Travis J (1997) Biosynthesis of alpha1-proteinase inhibitor by human lung-derived epithelial cells. J Biol Chem 272:8250–8255CrossRefPubMedGoogle Scholar
  8. 8.
    Ikari Y, Mulvihill E, Schwartz SM (2001) Alpha 1-Proteinase inhibitor, alpha 1-antichymotrypsin, and alpha 2-macroglobulin are the antiapoptotic factors of vascular smooth muscle cells. J Biol Chem 276:11798–11803CrossRefPubMedGoogle Scholar
  9. 9.
    Daemen MA, Heemskerk VH, van’t Veer C, Denecker G, TG W, Vandenabeele P, WA B (2000) Functional protection by acute phase proteins alpha(1)-acid glycoprotein and alpha(1)-antitrypsin against ischemia/reperfusion injury by preventing apoptosis and inflammation. Circulation 102:1420–1426CrossRefPubMedGoogle Scholar
  10. 10.
    Petrache I, Fijalkowska I, Zhen L, Medler TR, Brown E, Cruz P, Choe KH, Taraseviciene-Stewart L, Scerbavicius R, Shapiro L, Zhang B, Song S, Hicklin D, Voelkel NF, Flotte T, Tuder RM (2006) A novel antiapoptotic role for alpha1-antitrypsin in the prevention of pulmonary emphysema. Am J Respir Crit Care Med 173:1222–1228CrossRefPubMedPubMedCentralGoogle Scholar
  11. 11.
    Greene CM, Miller SD, Carroll TP, Oglesby IK, Ahmed F, O’Mahony M, Taggart CC, McElvaney NG, O’Neill SJ (2010) Anti-apoptotic effects of Z alpha1-antitrypsin in human bronchial epithelial cells. Eur Respir J 35:1155–1163CrossRefPubMedGoogle Scholar
  12. 12.
    Petrache I, Fijalkowska I, Medler TR, Skirball J, Cruz P, Zhen L, Petrache HI, Flotte TR, Tuder RM (2006) Alpha-1 antitrypsin inhibits caspase-3 activity, preventing lung endothelial cell apoptosis. Am J Pathol 169:1155–1166CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Lockett AD, Van Demark M, Gu Y, Schweitzer KS, Sigua N, Kamocki K, Fijalkowska I, Garrison J, Fisher AJ, Serban K, Wise RA, Flotte TR, Mueller C, RG P Jr, Petrache HI, Tuder RM, Petrache I (2012) Effect of cigarette smoke exposure and structural modifications on the α-1 antitrypsin interaction with caspases. Mol Med 18:445–454CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Higashiyama M, Doi O, Kodama K, Yokouchi H, Tateishi R (1992) An evaluation of the prognostic significance of alpha-1-antitrypsin expression in adenocarcinomas of the lung: an immunohistochemical analysis. Br J Cancer 65:300–302CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Kwon CH, Park HJ, Lee JR, Kim HK, Jeon TY, Jo HJ, Kim DH, Kim GH, Park DY (2014) Serpin peptidase inhibitor clade a member 1 is a biomarker of poor prognosis in gastric cancer. Br J Cancer 111:1993–2002CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Chang YH, Lee SH, Liao IC, Huang SH, Cheng HC, Liao PC (2012) Secretomic analysis identifies alpha-1 antitrypsin (A1AT) as a required protein in cancer cell migration, invasion, and pericellular fibronectin assembly for facilitating lung colonization of lung adenocarcinoma cells. Mol Cell Proteomics 11:1320–1339CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Kataoka H, Uchino H, Iwamura T, Seiki M, Nabeshima K, Koono M (1999) Enhanced tumor growth and invasiveness in vivo by a carboxyl-terminal fragment of alpha1-proteinase inhibitor generated by matrix metalloproteinases: a possible modulatory role in natural killer cytotoxicity. Am J Pathol 154:457–468CrossRefPubMedPubMedCentralGoogle Scholar
  18. 18.
    Zelvyte I, Lindgren S, Janciauskiene S (2003) Multiple effects of alpha1-antitrypsin on breast carcinoma MDA-MB 468 cell growth and invasiveness. Eur J Cancer Prev 12:117–124CrossRefPubMedGoogle Scholar
  19. 19.
    Zhao W, Yang Z, Liu X, Tian Q, Lv Y, Liang Y, Li C, Gao X, Chen L (2013) Identification of α1-antitrypsin as a potential prognostic biomarker for advanced nonsmall cell lung cancer treated with epidermal growth factor receptor tyrosine kinase inhibitors by proteomic analysis. J Int Med Res 41:573–583CrossRefPubMedGoogle Scholar
  20. 20.
    Topic A, Ljujic M, Petrovic-Stanojevic N, Dopudja-Pantic V, Radojkovic D (2011) Phenotypes and serum level of alpha-1-antitrypsin in lung cancer. J BUON 16:672–676PubMedGoogle Scholar
  21. 21.
    Varela AS, López Sáez JJ (1995) Utility of plasmatic levels of alpha-1-antiprotease (A1AP) as a cancer marker. Cancer Lett 89:15–21CrossRefPubMedGoogle Scholar
  22. 22.
    Pérez-Holanda S, Blanco I, Menéndez M, Rodrigo L (2014) Serum concentration of alpha-1 antitrypsin is significantly higher in colorectal cancer patients than in healthy controls. BMC Cancer 14:355CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Mijatovic S, Maksimovic-Ivanic D, Radovic J, Popadic D, Momcilovic M, Harhaji L, Miljkovic D, Trajkovic V (2004) Aloe-emodin prevents cytokine-induced tumor cell death: the inhibition of auto-toxic nitric oxide release as a potential mechanism. Cell Mol Life Sci 61:1805–1815CrossRefPubMedGoogle Scholar
  24. 24.
    McCubrey JA, Steelman LS, Abrams SL, Misaghian N, Chappell WH, Basecke J, Nicoletti F, Libra M, Ligresti G, Stivala F, Maksimovic-Ivanic D, Mijatovic S, Montalto G, Cervello M, Laidler P, Bonati A, Evangelisti C, Cocco L, Martelli AM (2012) Targeting the cancer initiating cell: the ultimate target for cancer therapy. Curr Pharm Des 18:1784–1795CrossRefPubMedGoogle Scholar
  25. 25.
    EMBL-EBI Expression Atlas, available on:
  26. 26.
    Kubisch R, Meissner L, Krebs S, Blum H, Günther M, Roidl A, Wagner EA (2013) Comprehensive gene expression analysis of resistance formation upon metronomic cyclophosphamide therapy. Transl Oncol 6:1–9CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    El-Akawi ZJ, Abu-Awad AM, Sharara AM, Khader Y (2010) The importance of alpha-1 antitrypsin (alpha1-AT) and neopterin serum levels in the evaluation of non-small cell lung and prostate cancer patients. Neuroendocrinol Lett 31:113–116PubMedGoogle Scholar
  28. 28.
    Zelvyte I, Wallmark A, Piitulainen E, Westin U, Janciauskiene S (2004) Increased plasma levels of serine proteinase inhibitors in lung cancer patients. Anticancer Res 24:241–247PubMedGoogle Scholar
  29. 29.
    Li Y, Krowka MJ, Qi Y, Katzmann JA, Song Y, Mandrekar SJ, Yang P (2011) Alpha1-antitrypsin deficiency carriers, serum alpha 1-antitrypsin concentration, and non-small cell lung cancer survival. J Thorac Oncol 6:291–295CrossRefPubMedPubMedCentralGoogle Scholar
  30. 30.
    Normandin K, Péant B, Le Page C, de Ladurantaye M, Ouellet V, Tonin PN, Provencher DM, Mes-Masson AM (2010) Protease inhibitor SERPINA1 expression in epithelial ovarian cancer. Clin Exp Metastasis 27:55–69CrossRefPubMedGoogle Scholar
  31. 31.
    Zelvyte I, Ohlsson B, Axelson J, Janciauskiene S (2003) Diverse responses between human pancreatic cancer cell lines to native alpha 1-antitrypsin and its C-terminal fragment. Anticancer Res 23(3B):2267–2273PubMedGoogle Scholar
  32. 32.
    Zelvyte I, Sjögren HO, Janciauskiene S (2002) Effects of native and cleaved forms of alpha1-antitrypsin on ME 1477 tumor cell functional activity. Cancer Detect Prev 26:256–265CrossRefPubMedGoogle Scholar
  33. 33.
    Boulikas T, Vougiouka M (2003) Cisplatin and platinum drugs at the molecular level. Oncol Rep 10:1663–1682PubMedGoogle Scholar
  34. 34.
    Fuertes MA, Castilla J, Alonso C, Pérez JM (2003) Cisplatin biochemical mechanism of action: from cytotoxicity to induction of cell death through interconnections between apoptotic and necrotic pathways. Curr Med Chem 10:257–266CrossRefPubMedGoogle Scholar
  35. 35.
    Ljujic M, Mijatovic S, Bulatovic MZ, Mojic M, Maksimovic-Ivanic D, Radojkovic D, Topic A (2016) Alpha-1 antitrypsin affects U0126-induced cytotoxicity in colon cancer cell line (HCT116. Mol Biol 50:153–156CrossRefGoogle Scholar
  36. 36.
    Pattingre S, Tassa A, Qu X, Garuti R, Liang XH, Mizushima N, Packer M, Schneider MD, Levine B (2005) Bcl-2 antiapoptotic proteins inhibit Beclin 1-dependent autophagy. Cell 122:927–939CrossRefPubMedGoogle Scholar
  37. 37.
    Marquez RT, Xu L (2012) Bcl-2:Beclin 1 complex: multiple, mechanisms regulating autophagy/apoptosis toggle switch. Am J Cancer Res 2:214–221PubMedPubMedCentralGoogle Scholar
  38. 38.
    Wei Y, Pattingre S, Sinha S, Bassik M, Levine B (2008) JNK1-mediated phosphorylation of Bcl-2 regulates starvation-induced autophagy. Mol Cell 30:678–688CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Chekhun VF, Kovtonyuk OV, Todor IN, Kulik GI (2005) Total proteolytic activity and levels of the main proteinase inhibitors in blood plasma of mice bearing Lewis lung carcinoma upon development of resistance to cisplatin. Exp Oncol 27:286–289PubMedGoogle Scholar
  40. 40.
    Dabbagh K, Laurent GJ, Shock A, Leoni P, Papakrivopoulou J, Chambers RC (2001) Alpha-1-antitrypsin stimulates fibroblast proliferation and procollagen production and activates classical MAP kinase signalling pathways. J Cell Physiol 186:73–81CrossRefPubMedGoogle Scholar
  41. 41.
    Gonias SL, Swaim MW, Massey MF, Pizzo SV (1988) cis-dichlorodiammineplatinum (II) as a selective modifier of the oxidation-sensitive reactive-center methionine in alpha 1-antitrypsin. J Biol Chem 263:393–397PubMedGoogle Scholar
  42. 42.
    Kloth JN, Gorter A, Fleuren GJ, Oosting J, Uljee S, ter Haar N, Dreef EJ, Kenter GG, Jordanova ES (2008) Elevated expression of SerpinA1 and SerpinA3 in HLA-positive cervical carcinoma. J Pathol 215:222–230CrossRefPubMedGoogle Scholar
  43. 43.
    Kwon CH, Park HJ, Choi JH, Lee JR, Kim HK, Jo HJ, Kim HS, Oh N, Song GA, do Y P (2015) Snail and serpinA1 promote tumor progression and predict prognosis in colorectal cancer. Oncotarget 6:20312–20326CrossRefPubMedPubMedCentralGoogle Scholar
  44. 44.
    JY W, Cheng CC, Wang JY, DC W, Hsieh JS, Lee SC, Wang WM (2014) Discovery of tumor markers for gastric cancer by proteomics. PLoS One 9:e84158CrossRefGoogle Scholar
  45. 45.
    Goodarzi MT, Turner GA (1995) Decreased branching, increased fucosylation and changed sialylation of alpha-1-proteinase inhibitor in breast and ovarian cancer. Clin Chim Acta 236:161–171CrossRefPubMedGoogle Scholar
  46. 46.
    Comunale MA, Rodemich-Betesh L, Hafner J, Wang M, Norton P, Di Bisceglie AM, Block T, Mehta A (2010) Linkage specific fucosylation of alpha-1-antitrypsin in liver cirrhosis and cancer patients: implications for a biomarker of hepatocellular carcinoma. PLoS One 5:e12419CrossRefPubMedPubMedCentralGoogle Scholar
  47. 47.
    Wang Y, Kuramitsu Y, Yoshino S, Takashima M, Zhang X, Ueno T, Suzuki N, Oka M, Nakamura K (2011) Screening for serological biomarkers of pancreatic cancer by two-dimensional electrophoresis and liquid chromatography-tandem mass spectrometry. Oncol Rep 26:287–292PubMedGoogle Scholar
  48. 48.
    Grivennikov SI, Greten FR, Karin M (2010) Immunity, inflammation, and cancer. Cell 140:883–899CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Huang W, Ding X, Li B, Fan M, Zhou T, Sun H, Yi Y, Zhang J (2013) Serum biomarkers analyzed by LC-MS/MS as predictors for short outcome of non-small cell lung cancer patients treated with chemoradiotherapy. Neoplasma 60:11–18CrossRefPubMedGoogle Scholar
  50. 50.
    Woźniak B, Mila-Kierzenkowska C, WoŸniak A, Drewa G, Sopońska M, Drewa T, Krzyzyńska-Malinowska E, Makarewicz R, Kowalski T, Szmytkowska K (2008) The effect of combined therapy on activity of cathepsin D and alpha-1-antitrypsin in the blood serum of women with cervical cancer. Eur J Gynaecol Oncol 29:617–619PubMedGoogle Scholar

Copyright information

© Arányi Lajos Foundation 2016

Authors and Affiliations

  • Mila Ljujic
    • 1
  • Sanja Mijatovic
    • 2
  • Mirna Z. Bulatovic
    • 2
  • Marija Mojic
    • 2
  • Danijela Maksimovic-Ivanic
    • 2
  • Dragica Radojkovic
    • 1
  • Aleksandra Topic
    • 3
    Email author
  1. 1.Institute of Molecular Genetics and Genetic EngineeringUniversity of BelgradeBelgradeSerbia
  2. 2.Institute for Biological Research “Sinisa Stankovic”University of BelgradeBelgradeSerbia
  3. 3.Department of Medical Biochemistry, Faculty of PharmacyUniversity of BelgradeBelgradeSerbia

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