Abstract
Carbonic anhydrase IX (CAIX) is a strictly membranous expressed metalloenzyme involved in cell adhesion, pH homeostasis, and cancer progression. The protein is specifically overexpressed in a wide variety of malignant tumors. This study was designed to assess the role of CAIX in primary vulvar cancer. One hundred forty-two well-characterized primary vulvar carcinomas were analyzed on a tissue microarray (TMA). Three tissue cores were sampled from each tumor. CAIX expression was determined by immunohistochemistry, using a four-step scoring system. To determine CAIX expression in benign vulvar tissue, we constructed a TMA with 120 samples of normal mucosa and non-neoplastic diseases. CAIX expression was found in 77/135 (57%) of all assessable vulvar cancer specimens and 48 (35.5%) exhibited a moderate or strong expression. CAIX expression in vulvar carcinomas was significantly stronger compared to non-neoplastic vulvar tissue (p < 0.001). High levels of CAIX expression were related to pT stage (p < 0.01), tumor size (p < 0.01), depth of invasion (p < 0.05), as well as inguinal lymph node metastases (p < 0.05). There was also a trend towards shorter recurrence-free patient survival in CAIX-positive compared to CAIX-negative vulvar cancers. CAIX staining results in different tissue cores from the same tumor were homogeneous, raising the possibility of a hypoxia-independent expression. In conclusion, CAIX is overexpressed in the majority of vulvar carcinomas with relationships to advanced tumor stages and development of lymph node metastases. Our data support the potential therapeutic benefit of newly developed targeting antibodies in advanced vulvar cancer.
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Vander Heiden MG, Cantley LC, Thompson CB (2009) Understanding the Warburg effect: the metabolic requirements of cell proliferation. Science 324:1029–1033
Fang JS, Gillies RD, Gatenby RA (2008) Adaptation to hypoxia and acidosis in carcinogenesis and tumor progression. Semin Cancer Biol 18:330–337
Potter CP, Harris AL (2003) Diagnostic, prognostic and therapeutic implications of carbonic anhydrases in cancer. Br J Cancer 89:2–7
Winum JY, Poulsen SA, Supuran CT (2009) Therapeutic applications of glycosidic carbonic anhydrase inhibitors. Med Res Rev 29:419–435
Zavada J, Zavadova Z, Pastorekova S et al (1993) Expression of MaTu-MN protein in human tumor cultures and in clinical specimens. Int J Cancer 54:268–274
Potter C, Harris Al (2004) Hypoxia inducible carbonic anhydrase IX, marker of tumour hypoxia, survival pathway and therapy target. Cell Cycle 3:164–167
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
Kaluz S, Kaluzova M, Liao SY et al (2009) Transcriptional control of the tumor- and hypoxia-marker carbonic anhydrase 9: a one transcription factor (HIF-1) show? Biochim Biophys Acta 1795:162–172
Ivanov S, Liao SY, Ivanova A et al (2001) Expression of hypoxia-inducible cell-surface transmembrane carbonic anhydrases in human cancer. Am J Pathol 158:905–919
Hussain SA, Ganesan R, Reynolds G et al (2007) Hypoxia-regulated carbonic anhydrase IX expression is associated with poor survival in patients with invasive breast cancer. Br J Cancer 96:104–109
Klatte T, Seligson DB, Rao JY (2009) Carbonic anhydrase IX in bladder cancer: a diagnostic, prognostic, and therapeutic molecular marker. Cancer 115:1448–1458
Loncaster JA, Harris AL, Davidson SE et al (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 61:6394–6399
Swinson DE, Jones JL, Richardson D (2003) Carbonic anhydrase IX expression, a novel surrogate marker of tumor hypoxia, is associated with a poor prognosis in non-small-cell lung cancer. J Clin Oncol 21:473–482
Simon R, Mirlacher M, Sauter G (2004) Tissue microarrays. Biotechniques 36:98–105
Petersen I, Schewe C, Schluns K et al (2007) Inter-laboratory validation of PCR-based HPV detection in pathology specimens. Virchows Arch 451:701–716
Landis JR, Koch GG (1977) The measurement of observer agreement for categorical data. Biometrics 33:159–174
Woelber L, Mahner S, Voelker K et al (2009) Clinicopathological prognostic factors and patterns of recurrence in vulvar cancer. Anticancer Res 29:545–552
Lee WY, Huang SC, Hsu KF et al (2008) Roles for hypoxia-regulated genes during cervical carcinogenesis: somatic evolution during the hypoxia-glycolysis-acidosis sequence. Gynecol Oncol 108:377–384
Choi SW, Kim JY, Park JY et al (2008) Expression of carbonic anhydrase IX is associated with postoperative recurrence and poor prognosis in surgically treated oral squamous cell carcinoma. Hum Pathol 39:1317–1322
Tanaka N, Kato H, Inose T et al (2008) Expression of carbonic anhydrase 9, a potential intrinsic marker of hypoxia, is associated with poor prognosis in oesophageal squamous cell carcinoma. Br J Cancer 99:1468–1475
Bubendorf L, Nocito A, Moch H et al (2001) Tissue microarray (TMA) technology: miniaturized pathology archives for high-throughput in situ studies. J Pathol 195:72–79
Ruiz C, Seibt S, Al Kuraya K (2006) Tissue microarrays for comparing molecular features with proliferation activity in breast cancer. Int J Cancer 118:2190–2194
Torhorst J, Bucher C, Kononen J et al (2001) Tissue microarrays for rapid linking of molecular changes to clinical endpoints. Am J Pathol 159:2249–2256
Fons G, Hasibuan SM, van der Velden J et al (2007) Validation of tissue microarray technology in endometrioid cancer of the endometrium. J Clin Pathol 60:500–503
Gulmann C, Butler D, Kay E et al (2003) Biopsy of a biopsy: validation of immunoprofiling in gastric cancer biopsy tissue microarrays. Histopathology 42:70–76
Rubin MA, Dunn R, Strawderman M et al (2002) Tissue microarray sampling strategy for prostate cancer biomarker analysis. Am J Surg Pathol 26:312–319
Fons G, van der Velden J, Burger M et al (2009) Validation of tissue microarray technology in vulvar cancer. Int J Gynecol Pathol 28:76–82
Ruan K, Song G, Ouyang G (2009) Role of hypoxia in the hallmarks of human cancer. J Cell Biochem 107:1053–1062
Kaluz S, Kaluzova M, Chrastina A et al (2002) Lowered oxygen tension induces expression of the hypoxia marker MN/carbonic anhydrase IX in the absence of hypoxia-inducible factor 1 alpha stabilization: a role for phosphatidylinositol 3'-kinase. Cancer Res 62:4469–4477
Kopacek J, Barathova M, Dequiedt F et al (2005) MAPK pathway contributes to density- and hypoxia-induced expression of the tumor-associated carbonic anhydrase IX. Biochim Biophys Acta 1729:41–49
Denko NC (2008) Hypoxia, HIF1 and glucose metabolism in the solid tumour. Nat Rev Cancer 8:705–713
Kong CS (2009) The relationship between human papillomavirus status and other molecular prognostic markers in head and neck squamous cell carcinomas. Int J Radiat Oncol Biol Phys 74:553–561
Resnick M (1996) Viral and histopathologic correlates of MN and MIB-1 expression in cervical intraepithelial neoplasia. Hum Pathol 27:234–239
Wykoff CC, Beasley N, Watson PH et al (2001) Expression of the hypoxia-inducible and tumor-associated carbonic anhydrases in ductal carcinoma in situ of the breast. Am J Pathol 158:1011–1019
Drew PA, al-Abbadi MA, Orlando CA (1996) Prognostic factors in carcinoma of the vulva: a clinicopathologic and DNA flow cytometric study. Int J Gynecol Pathol 15:235–241
Svastova E, Hulikova A, Rafajova M et al (2004) Hypoxia activates the capacity of tumor-associated carbonic anhydrase IX to acidify extracellular pH. FEBS Lett 577:439–445
Swietach P, Patiar S, Supuran CT et al (2009) The role of carbonic anhydrase 9 in regulating extracellular and intracellular pH in 3-D tumor-cell growths. J Biol Chem 284:20299–20310
Mulders P, Bleumer I, Debruyne F, et al. (2004) Specific monoclonal antibody-based immunotherapy by targeting the RCC-associated antigen carbonic anhydrase-IX(G250/MN) Urologe A 43 Suppl 3:S146-147: 146-147
Bauer S, Oosterwijk-Wakka JC, Adrian N et al (2009) Targeted therapy of renal cell carcinoma: synergistic activity of cG250-TNF and IFNg. Int J Cancer 125:115–123
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Matthias Choschzick and Linn Woelber contributed equally to this study.
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Choschzick, M., Woelber, L., Hess, S. et al. Overexpression of carbonic anhydrase IX (CAIX) in vulvar cancer is associated with tumor progression and development of locoregional lymph node metastases. Virchows Arch 456, 483–490 (2010). https://doi.org/10.1007/s00428-010-0905-9
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DOI: https://doi.org/10.1007/s00428-010-0905-9