Abstract
Background
Carbonic anhydrase IX (CAIX) is involved in pH homeostasis, growth and survival of tumor cells. Besides the membranous form of CAIX, a soluble form is detectable in serum (s-CAIX). Overexpression of CAIX in tumors offers the opportunity for therapeutic strategies such as CAIX targeting antibodies. The aim of this study was to examine the relationships of CAIX mRNA expression and s-CAIX levels with clinicopathological parameters and survival of patients with primary breast cancer.
Methods
Tumor tissue of 169 primary breast cancer patients was analyzed for RNA expression by microarray analysis (Affymetrix HG-U133A). Concentration of s-CAIX was determined by ELISA in blood samples of 140 patients.
Results
In tumor tissue, CAIX mRNA signal intensities (MAS5 values) ranged from 34 to 2,513. Higher CAIX expression was associated with younger age (</≥50 years p = 0.040), negative hormone receptors (estrogen receptor p = 0.004; progesterone receptor p = 0.022) and positive nodal status (p = 0.001) as well as with shorter disease-free survival (p = 0.031). Concentrations of s-CAIX ranged from 56 to 1,500 pg/ml. There was no correlation between s-CAIX and CAIX mRNA levels as well as clinicopathological characteristics or outcome.
Conclusion
In contrast to reported immunohistochemical studies, we performed RNA-based tissue analyses of CAIX expression and, for the first time, analyzed CAIX serum levels in primary breast cancer. The correlations between CAIX RNA expression and prognostic factors and patient outcome support a biologic role of CAIX in early breast cancer. A role of s-CAIX in primary breast cancer is not supported by our findings.
Similar content being viewed by others
References
Bareschino MA, Schettino C, Colantuoni G, Rossi E, Rossi A, Maione P, Ciardiello F, Gridelli C (2011) The role of antiangiogenetic agents in the treatment of breast cancer. Curr Med Chem 18(33):5022–5032
Bleumer I, Oosterwijk E, Oosterwijk-Wakka JC, Voller MC, Melchior S, Warnaar SO, Mala C, Beck J, Mulders PF (2006) A clinical trial with chimeric monoclonal antibody WX-G250 and low dose interleukin-2 pulsing scheme for advanced renal cell carcinoma. J Urol 175(1):57–62. doi:10.1016/S0022-5347(05)00040-6
Brennan DJ, Jirstrom K, Kronblad A, Millikan RC, Landberg G, Duffy MJ, Ryden L, Gallagher WM, O’Brien SL (2006) CA IX is an independent prognostic marker in premenopausal breast cancer patients with one to three positive lymph nodes and a putative marker of radiation resistance. Clin Cancer Res 12(21):6421–6431. doi:10.1158/1078-0432.CCR-06-0480
Bui MH, Seligson D, Han KR, Pantuck AJ, Dorey FJ, Huang Y, Horvath S, Leibovich BC, Chopra S, Liao SY, Stanbridge E, Lerman MI, Palotie A, Figlin RA, Belldegrun AS (2003) Carbonic anhydrase IX is an independent predictor of survival in advanced renal clear cell carcinoma: implications for prognosis and therapy. Clin Cancer Res 9(2):802–811
Carney WP (2007) Circulating oncoproteins HER2/neu, EGFR and CAIX (MN) as novel cancer biomarkers. Expert Rev Mol Diagn 7(3):309–319. doi:10.1586/14737159.7.3.309
Chia SK, Wykoff CC, Watson PH, Han C, Leek RD, Pastorek J, Gatter KC, Ratcliffe P, Harris AL (2001) Prognostic significance of a novel hypoxia-regulated marker, carbonic anhydrase IX, in invasive breast carcinoma. J Clin Oncol 19(16):3660–3668
Choschzick M, Oosterwijk E, Muller V, Woelber L, Simon R, Moch H, Tennstedt P (2011) Overexpression of carbonic anhydrase IX (CAIX) is an independent unfavorable prognostic marker in endometrioid ovarian cancer. Virchows Arch 459(2):193–200. doi:10.1007/s00428-011-1105-y
Duffy MJ, Evoy D, McDermott EW (2010) CA 15–3: uses and limitation as a biomarker for breast cancer. Clin Chim Acta 411(23–24):1869–1874. doi:10.1016/j.cca.2010.08.039
Fang JS, Gillies RD, Gatenby RA (2008) Adaptation to hypoxia and acidosis in carcinogenesis and tumor progression. Semin Cancer Biol 18(5):330–337. doi:10.1016/j.semcancer.2008.03.011
Gatenby RA, Smallbone K, Maini PK, Rose F, Averill J, Nagle RB, Worrall L, Gillies RJ (2007) Cellular adaptations to hypoxia and acidosis during somatic evolution of breast cancer. Br J Cancer 97(5):646–653. doi:10.1038/sj.bjc.6603922
Hayes DF, Ethier S, Lippman ME (2006) New guidelines for reporting of tumor marker studies in breast cancer research and treatment: remark. Breast Cancer Res Treat 100(2):237–238. doi:10.1007/s10549-006-9253-5
Hussain SA, Ganesan R, Reynolds G, Gross L, Stevens A, Pastorek J, Murray PG, Perunovic B, Anwar MS, Billingham L, James ND, Spooner D, Poole CJ, Rea DW, Palmer DH (2007) Hypoxia-regulated carbonic anhydrase IX expression is associated with poor survival in patients with invasive breast cancer. Br J Cancer 96(1):104–109. doi:10.1038/sj.bjc.6603530
Ivanov S, Liao SY, Ivanova A, Danilkovitch-Miagkova A, Tarasova N, Weirich G, Merrill MJ, Proescholdt MA, Oldfield EH, Lee J, Zavada J, Waheed A, Sly W, Lerman MI, Stanbridge EJ (2001) Expression of hypoxia-inducible cell-surface transmembrane carbonic anhydrases in human cancer. Am J Pathol 158(3):905–919. doi:10.1016/S0002-9440(10)64038-2
Kaluz S, Kaluzova M, Liao SY, Lerman M, Stanbridge EJ (2009) Transcriptional control of the tumor- and hypoxia-marker carbonic anhydrase 9: a one transcription factor (HIF-1) show? Biochim Biophys Acta 1795(2):162–172. doi:10.1016/j.bbcan.2009.01.001
Kock L, Mahner S, Choschzick M, Eulenburg C, Milde-Langosch K, Schwarz J, Jaenicke F, Muller V, Woelber L (2011) Serum carbonic anhydrase IX and its prognostic relevance in vulvar cancer. Int J Gynecol Cancer 21(1):141–148. doi:10.1097/IGC.0b013e318204c34f
Li G, Feng G, Gentil-Perret A, Genin C, Tostain J (2008) Serum carbonic anhydrase 9 level is associated with postoperative recurrence of conventional renal cell cancer. J Urol 180(2):510–513. doi:10.1016/j.juro.2008.04.024 discussion 513–514
Loncaster JA, Harris AL, Davidson SE, Logue JP, Hunter RD, Wycoff CC, Pastorek J, Ratcliffe PJ, Stratford IJ, West CM (2001) Carbonic anhydrase (CA IX) expression, a potential new intrinsic marker of hypoxia: correlations with tumor oxygen measurements and prognosis in locally advanced carcinoma of the cervix. Cancer Res 61(17):6394–6399
Lou Y, McDonald PC, Oloumi A, Chia S, Ostlund C, Ahmadi A, Kyle A, Auf dem Keller U, Leung S, Huntsman D, Clarke B, Sutherland BW, Waterhouse D, Bally M, Roskelley C, Overall CM, Minchinton A, Pacchiano F, Carta F, Scozzafava A, Touisni N, Winum JY, Supuran CT, Dedhar S (2011) Targeting tumor hypoxia: suppression of breast tumor growth and metastasis by novel carbonic anhydrase IX inhibitors. Cancer Res 71(9):3364–3376. doi:10.1158/0008-5472.CAN-10-4261
Mori I, Yang Q, Kakudo K (2002) Predictive and prognostic markers for invasive breast cancer. Pathol Int 52(3):186–194
Müller V, Riethdorf S, Rack B, Janni W, Fasching PA, Solomayer E, Aktas B, Kasimir-Bauer S, Zeitz J, Pantel K, Fehm T (2011) Prospective evaluation of serum tissue inhibitor of metalloproteinase 1 and carbonic anhydrase IX in correlation to circulating tumor cells in patients with metastatic breast cancer. Breast Cancer Res 13(4):R71. doi:10.1186/bcr2916
Potter C, Harris AL (2004) Hypoxia inducible carbonic anhydrase IX, marker of tumour hypoxia, survival pathway and therapy target. Cell Cycle 3(2):164–167
Potter CP, Harris AL (2003) Diagnostic, prognostic and therapeutic implications of carbonic anhydrases in cancer. Br J Cancer 89(1):2–7. doi:10.1038/sj.bjc.6600936
Robertson N, Potter C, Harris AL (2004) Role of carbonic anhydrase IX in human tumor cell growth, survival, and invasion. Cancer Res 64(17):6160–6165. doi:10.1158/0008-5472.CAN-03-2224
Schroder C, Witzel I, Muller V, Krenkel S, Wirtz RM, Janicke F, Schumacher U, Milde-Langosch K (2011) Prognostic value of intercellular adhesion molecule (ICAM)-1 expression in breast cancer. J Cancer Res Clin Oncol 137(8):1193–1201. doi:10.1007/s00432-011-0984-2
Siebels M, Rohrmann K, Oberneder R, Stahler M, Haseke N, Beck J, Hofmann R, Kindler M, Kloepfer P, Stief C (2011) A clinical phase I/II trial with the monoclonal antibody cG250 (RENCAREX(R)) and interferon-alpha-2a in metastatic renal cell carcinoma patients. World J Urol 29(1):121–126. doi:10.1007/s00345-010-0570-2
Stoner M, Saville B, Wormke M, Dean D, Burghardt R, Safe S (2002) Hypoxia induces proteasome-dependent degradation of estrogen receptor alpha in ZR-75 breast cancer cells. Mol Endocrinol 16(10):2231–2242
Tafreshi NK, Bui MM, Bishop K, Lloyd MC, Enkemann SA, Lopez AS, Abrahams D, Carter BW, Vagner J, Gobmyer SR, Gillies RJ, Morse DL (2012) Noninvasive detection of breast cancer lymph node metastasis using carbonic anhydrases IX and XII targeted imaging probes. Clin Cancer Res 18(1):207–219. doi:10.1158/1078-0432.CCR-11-0238
Winum JY, Poulsen SA, Supuran CT (2009a) Therapeutic applications of glycosidic carbonic anhydrase inhibitors. Med Res Rev 29(3):419–435. doi:10.1002/med.20141
Winum JY, Scozzafava A, Montero JL, Supuran CT (2009b) Inhibition of carbonic anhydrase IX: a new strategy against cancer. Anti-Cancer Agents Med Chem 9(6):693–702
Woelber L, Kress K, Kersten JF, Choschzick M, Kilic E, Herwig U, Lindner C, Schwarz J, Jaenicke F, Mahner S, Milde-Langosch K, Mueller V, Ihnen M (2011) Carbonic anhydrase IX in tumor tissue and sera of patients with primary cervical cancer. BMC cancer 11:12. doi:10.1186/1471-2407-11-12
Woelber L, Mueller V, Eulenburg C, Schwarz J, Carney W, Jaenicke F, Milde-Langosch K, Mahner S (2010) Serum carbonic anhydrase IX during first-line therapy of ovarian cancer. Gynecol Oncol 117(2):183–188. doi:10.1016/j.ygyno.2009.11.029
Zavada J, Zavadova Z, Pastorekova S, Ciampor F, Pastorek J, Zelnik V (1993) Expression of MaTu-MN protein in human tumor cultures and in clinical specimens. Int J Cancer 54(2):268–274
Zavada J, Zavadova Z, Zat’ovicova M, Hyrsl L, Kawaciuk I (2003) Soluble form of carbonic anhydrase IX (CA IX) in the serum and urine of renal carcinoma patients. Br J Cancer 89(6):1067–1071. doi:10.1038/sj.bjc.6601264
Acknowledgments
We thank Katrin Beck and Maila Rossberg for excellent technical assistance.
Conflict of interest
All authors declare no competing interests. ELISA kits were provided at no cost by Oncogene Science, a former part of Siemens Medical Solutions Diagnostics and now part of Wilex. The company did not have a role in study design or collection, analysis and interpretation of data nor in the writing of the manuscript.
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Schütze, D., Milde-Langosch, K., Witzel, I. et al. Relevance of cellular and serum carbonic anhydrase IX in primary breast cancer. J Cancer Res Clin Oncol 139, 747–754 (2013). https://doi.org/10.1007/s00432-013-1378-4
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s00432-013-1378-4