Advertisement

Biologia

, Volume 73, Issue 6, pp 621–628 | Cite as

In vitro effect of carbonic anhydrase inhibitor acetazolamide on cell viability, migration and colony formation of colorectal cancer cells

  • Fuat Karakuş
  • Ergül Eyol
  • Kadir Yılmaz
  • Songül Ünüvar
Original Article
  • 4 Downloads

Abstract

Acidification of extracellular medium in malignant tumors increases the invasive behaviors of cancer cells. In normal healthy tissues, acid production is catalyzed by carbonic anhydrases. Some of the carbonic anhydrase enzymes are overexpressed in certain types of cancer. The present study aimed to investigate the effect of acetazolamide, a potent carbonic anhydrase inhibitor, on in vitro cultivated cancer cells. Three different assays (MTT test, wound healing and clonogenic assay) were performed using human colorectal adenocarcinoma cells (SW620) to evaluate the suppressive effect of acetazolamide, on the colorectal cancer cells migration ability, colony formation and cell viability. The dose-dependent (1–1000 μM) reducing effect of acetazolamide on the cell viability was more significant within the first 48 h. This inhibitory effect of acetazolamide was found to be decreased at 72 h, and affects cells migration ability of cells at 24 and 48 h. Acetazolamide was observed to inhibit the cell viability, migration and colony formation ability of cells, depending on dose.

Keywords

Colorectal cancer Acetazolamide Carbonic anhydrase SW620 Aquaporins 

Abbreviations

ANOVA

analysis of variance

anti-VEGF

anti-vascular endothelial growth factor

AQP

aquaporin

CA

carbonic anhydrase

CO2

carbon dioxide

EDTA

ethylenediaminetetraacetic acid

ELISA

enzyme-linked immunosorbent assay

FBS

fetal bovine serum

5-FU

5-fluorouracil

HCO3

bicarbonate

H2CO3

carbonic acid

IC50

half maximal inhibitory concentration

MTT

3-[4,5-dimethylthiazol-2-yl] -2,5-diphenyltetrazolium bromide

PBS

phosphate-buffered saline

RPMI

Roswell Park Memorial Institute

SD

standard deviation

SPSS

Statistical Package for the Social Sciences

Notes

Acknowledgments

The study was supported by Inonu University Scientific Research Projects Coordination Unit with project number 2014/29. The authors would like to thank Prof. Dr. Martin R. Berger (The German Cancer Research Center, Heidelberg, Germany). We thank the anonymous Reviewers for their valuable comments.

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

References

  1. Andre N, Schmiegel W (2005) Chemoradiotherapy for colorectal cancer. Gut 54:1194–1202.  https://doi.org/10.1136/gut.2004.062745 CrossRefPubMedPubMedCentralGoogle Scholar
  2. Benga G (2012) The first discovered water channel protein, later called aquaporin 1: molecular characteristics, functions and medical implications. Mol Asp Med 33:518–534.  https://doi.org/10.1016/j.mam.2012.06.001 CrossRefGoogle Scholar
  3. Bin K, Shi-Peng Z (2011) Acetazolamide inhibits aquaporin-1 expression and colon cancer xenograft tumor growth. Hepatogastroenterology 58:1502–1516.  https://doi.org/10.5754/hge11154 CrossRefPubMedGoogle Scholar
  4. Carlin S, Khan N, Ku T, Longo VA, Larson SM, Smith-Jones PM (2010) Molecular targeting of carbonic anhydrase IX in mice with hypoxic HT29 colorectal tumor xenografts. PLoS One 5:e10857.  https://doi.org/10.1371/journal.pone.0010857 CrossRefPubMedPubMedCentralGoogle Scholar
  5. Cianchi F, Vinci MC, Supuran CT, Peruzzi B, De Giuli P, Fasolis G, Perigli G, Pastorekova S, Papucci L, Pini A, Masini E, Puccetti L (2010) Selective inhibition of carbonic anhydrase IX decreases cell proliferation and induces ceramide-mediated apoptosis in human cancer cells. J Pharmacol Exp Ther 334:710–719.  https://doi.org/10.1124/jpet.110.167270 CrossRefPubMedGoogle Scholar
  6. De Monte C, Carradori S, Secci D, D’Ascenzio M, Vullo D, Ceruso M, Supuran CT (2014) Cyclic tertiary sulfamates: selective inhibition of the tumor-associated carbonic anhydrases IX and XII by N- and O-substituted acesulfame derivatives. Eur J Med Chem 84:240–246.  https://doi.org/10.1016/j.ejmech.2014.07.014 CrossRefPubMedGoogle Scholar
  7. Eyol E, Murtaga A, Zhivkova-Galunska M, Georges R, Zepp M, Djandji D, Kleeff J, Berger MR, Adwan H (2012) Few genes are associated with the capability of pancreatic ductal adenocarcinoma cells to grow in the liver of nude rats. Oncol Rep 28:2177–2187.  https://doi.org/10.3892/or.2012.2049 CrossRefPubMedGoogle Scholar
  8. Kaleağasıoğlu F, Berger MR (2014) Differential effects of erufosine on proliferation, wound healing and apoptosis in colorectal cancer cell lines. Oncol Rep 31:1407–1416.  https://doi.org/10.3892/or.2013.2942 CrossRefPubMedGoogle Scholar
  9. Li XJ, Xiang Y, Ma B, Qi XQ (2007) Effects of acetazolamide combined with or without NaCO3 on suppressing neoplasm growth, metastasis and Aquaporin-1 (AQP1) protein expression. Int J Mol Sci 8:229–240.  https://doi.org/10.3390/i8030229 CrossRefPubMedCentralGoogle Scholar
  10. Liang CC, Park AY, Guan JL (2007) In vitro scratch assay: a convenient and inexpensive method or analysis of cell migration in vitro. Nat Protoc 2:329–333.  https://doi.org/10.1038/nprot.2007.30 CrossRefPubMedGoogle Scholar
  11. McCall KA, Huang CC, Fierke CA (2000) Function and mechanism of zinc metalloenzymes. J Nutr 130:14375–14465.  https://doi.org/10.1093/jn/130.5.1437S CrossRefGoogle Scholar
  12. Mohammadpour R, Safarian S, Ejeian F, Sheikholya-Lavasani Z, Abdolmohammadi MH, Sheinabi N (2014) Acetazolamide triggers death induced autophagy in T-47D breast cancer cells. Cell Biol Int 38:228–238.  https://doi.org/10.1002/cbin.10197 CrossRefPubMedGoogle Scholar
  13. Mokhtari RB, Kumar S, Islam SS, Yazdanpanah M, Adeli K, Cutz E, Yeger H (2013) Combination of carbonic anhydrase inhibitor, acetazolamide, and sulforaphane, reduces the viability and growth of bronchial carcinoid cell lines. BMC Cancer 13:378.  https://doi.org/10.1186/1471-2407-13-378 CrossRefPubMedPubMedCentralGoogle Scholar
  14. Nepali K, Sharma S, Sharma M, Bed PM, Dhar KL (2014) Rational approaches, design strategies, structure activity relationship and mechanistic insights for anticancer hybrids. Eur J Med Chem 77:422–487.  https://doi.org/10.1016/j.ejmech.2014.03.018 CrossRefPubMedGoogle Scholar
  15. Neri D, Supuran CT (2011) Interfering with pH regulation in tumours as a therapeutic strategy. Nat Rev Drug Discov 10:767–777.  https://doi.org/10.1038/nrd3554 CrossRefPubMedGoogle Scholar
  16. Nico B, Ribatti D (2010) Aquaporins in tumor growth and angiogenesis. Cancer Lett 294:135–138.  https://doi.org/10.1016/j.canlet.2010.02.005 CrossRefPubMedGoogle Scholar
  17. Nielsen DL, Palshof JA, Larsen FO, Jensen BV, Pfeiffer P (2014) A systematic review of salvage therapy to patients with metastatic colorectal cancer previously treated with fluorouracil, oxaliplatin and irinotecan +/− targeted therapy. Cancer Treat Rev 40:701–715.  https://doi.org/10.1016/j.ctrv.2014.02.006 CrossRefPubMedGoogle Scholar
  18. Parkkila S, Rajaniemi H, Parkkila AK, Kivela J, Waheed A, Pastorekova S, Pastorek J, Sly WS (2000) Carbonic anhydrase inhibitor suppresses invasion of renal cancer cells in vitro. Proc Natl Acad Sci U S A 97:2220–2224.  https://doi.org/10.1073/pnas.040554897 CrossRefPubMedPubMedCentralGoogle Scholar
  19. Pastorek J, Pastorekova S (2015) Hypoxia-induced carbonic anhydrase IX as a target for cancer therapy: from biology to clinical use. Semin Cancer Biol 31:52–64.  https://doi.org/10.1016/j.semcancer.2014.08.002 CrossRefPubMedGoogle Scholar
  20. Pastorekova S, Kopacek J, Pastorek J (2007) Carbonic anhydrase inhibitors and the management of cancer. Curr Top Med Chem 7:865–878.  https://doi.org/10.2174/156802607780636708 CrossRefPubMedGoogle Scholar
  21. Robertson N, Potter C, Harris AL (2004) Role of carbonic anhydrase IX in human tumor cell growth, survival, and invasion. Cancer Res 64:6160–6165.  https://doi.org/10.1158/0008-5472.CAN-03-2224 CrossRefPubMedGoogle Scholar
  22. Scozzafava A, Supuran CT (2000) Carbonic anhydrase and matrix metalloproteinase inhibitors: sulfonylated amino acid hydroxamates with MMP inhibitory properties act as efficient inhibitors of CA isozymes I, II, and IV, and N-hydroxysulfonamides inhibit both these zinc enzymes. J Med Chem 43:3677–3687.  https://doi.org/10.1021/jm000027t CrossRefPubMedGoogle Scholar
  23. Scozzafava A, Menabuoni L, Mincione F, Briganti F, Mincione G, Supuran CT (1999) Carbonic anhydrase inhibitors. synthesis of water-soluble, topically effective, intraocular pressure-lowering aromatic/heterocyclic sulfonamides containing cationic or anionic moieties: is the tail more important than the ring? J Med Chem 42:2641–2650.  https://doi.org/10.1021/jm9900523 CrossRefPubMedGoogle Scholar
  24. Shi X, Wu S, Yang Y, Tang L, Wang Y, Dong J, Lü B, Jiang G, Zhao W (2014) AQP5 silencing suppresses p 38 MAPK signaling and improves drug resistance in colon cancer cells. Tumour Biol 35:7035–7045.  https://doi.org/10.1007/s13277-014-1956-3 CrossRefPubMedGoogle Scholar
  25. Song CW, Griffin R, Park HJ. (2006). Influence of tumor pH on therapeutic response. B. Teicher (Ed.). Cancer drug discovery and development: Cancer drug resistance. USA: Humana Press Inc.  https://doi.org/10.1007/978-1-59745-035-5_2
  26. Spugnini EP, Sonveaux P, Stock C, Perez-Sayans M, De Milito A, Avnet S, Garcìa AG, Harguindey S, Fais S. (2015). Proton channels and exchangers in cancer. Biochim Biophys Acta 1848:2715–2726. doi: https://doi.org/10.1016/j.bbamem.2014.10.015
  27. Supuran CT (2008) Carbonic anhydrases: novel therapeutic applications for inhibitors and activators. Nat Rev Drug Discov 7:168–181.  https://doi.org/10.1038/nrd2467 CrossRefPubMedGoogle Scholar
  28. Supuran CT, Scozzafava A (2000a) Carbonic anhydrase inhibitors and their therapeutic potential. Expert Opin Ther Pat 10:575–600.  https://doi.org/10.1517/13543776.10.5.575 CrossRefGoogle Scholar
  29. Supuran CT, Scozzafava A (2000b) Carbonic anhydrase inhibitors--part 94. 1,3,4-thiadiazole-2-sulfonamidederivatives as antitumor agents? Eur J Med Chem 35:867–874.  https://doi.org/10.1016/S0223-5234(00)00169-0 CrossRefPubMedGoogle Scholar
  30. Supuran CT, Scozzafava A (2002) Applications of carbonic anhydrase inhibitors and activators in therapy. Expert Opin Ther Pat 12:217–242.  https://doi.org/10.1517/13543776.12.2.217 CrossRefGoogle Scholar
  31. Teicher BA, Liu SD, Liu JT, Holden SA, Herman TS (1993) A carbonic anhydrase inhibitor as a potential modulator of cancer therapies. Anticancer Res 13:1549–1556PubMedGoogle Scholar
  32. Tekedereli I, Alpay SN, Akar U, Yuca E, Ayugo-Rodriguez C, Han HD, Sood AK, Lopez-Berestein G, Ozpolat B (2013) Therapeutic silencing of Bcl-2 by systemically administered siRNA nanotherapeutics inhibits tumor growth by autophagy and apoptosis and enhances the efficacy of chemotherapy in orthotopic xenograft models of ER (−) and ER (+) breast cancer. Mol Ther–Nucleic Acids 10:e121.  https://doi.org/10.1038/mtna.2013.45 CrossRefGoogle Scholar
  33. Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J, Jemal A (2015) Global cancer statistics. CA Cancer J Clin 65:87–108.  https://doi.org/10.3322/caac.21262 CrossRefPubMedGoogle Scholar
  34. Xiang Y, Ma B, Li T, Yu HM, Li XJ (2002) Acetazolamide suppresses tumor metastasis and related protein expression in mice bearing Lewis lung carcinoma. Acta Pharmacol Sin 23:745–751PubMedGoogle Scholar
  35. Xiang Y, Ma B, Li T, Gao JW, Yu HM, Li XJ (2004) Acetazolamide inhibits aquaporin-1protein expression and angiogenesis. Acta Pharmacol Sin 25:812–816PubMedGoogle Scholar
  36. Yan C, Zhu Y, Zhang X, Chen X, Zheng W, Yang J (2014) Down-regulated aquaporin 5 inhibits proliferation and migration of human epithelial ovarian cancer 3AO cells. J Ovar Res 7:78.  https://doi.org/10.1186/s13048-014-0078-2 CrossRefGoogle Scholar
  37. Yoshida T, Hojo S, Sekine S, Sawada S, Okumura T, Nagata T, Shimada Y, Tsukada K (2013) Expression of aquaporin-1 is a poor prognostic factor for stage II and III colon cancer. Mol Clin Oncol 1:953–958.  https://doi.org/10.3892/mco.2013.165 CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Institute of Molecular Biology, Slovak Academy of Sciences 2018

Authors and Affiliations

  • Fuat Karakuş
    • 1
    • 2
  • Ergül Eyol
    • 1
  • Kadir Yılmaz
    • 3
  • Songül Ünüvar
    • 1
  1. 1.Department of Pharmaceutical Toxicology, Faculty of Pharmacyİnönü UniversityMalatyaTurkey
  2. 2.Department of Pharmaceutical Toxicology, Faculty of PharmacyEge UniversityİzmirTurkey
  3. 3.Department of Chemistry, Faculty of Arts and Scienceİnönü UniversityMalatyaTurkey

Personalised recommendations