Suppression of carbonic anhydrase IX leads to aberrant focal adhesion and decreased invasion of tumor cells

Radvak,Peter; Repic,Marko; Svastova,Eliska; Takacova,Martina; Csaderova,Lucia; Strnad,Hynek; Pastorek,Jaromir; Pastorekova,Silvia; Kopacek,Juraj

doi:10.3892/or.2013.2226

Suppression of carbonic anhydrase IX leads to aberrant focal adhesion and decreased invasion of tumor cells

Authors:
- Peter Radvak
- Marko Repic
- Eliska Svastova
- Martina Takacova
- Lucia Csaderova
- Hynek Strnad
- Jaromir Pastorek
- Silvia Pastorekova
- Juraj Kopacek
View Affiliations

Affiliations: Institute of Virology, Department of Molecular Medicine, Slovak Academy of Sciences, 845 05 Bratislava, Slovak Republic, Institute of Molecular Genetics, Academy of Sciences of the Czech Republic, 142 20 Prague 4, Czech Republic
Published online on: January 4, 2013 https://doi.org/10.3892/or.2013.2226
Pages: 1147-1153

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Carbonic anhydrase IX (CA IX) is a well-recognized hypoxia marker with promising diagnostic and therapeutic value. CA IX regulates the pH in hypoxic tumor cells and, thereby, contributes to microenvironmental acidosis and cell migration. To gain a better insight into the molecular processes driven by CA IX, we performed gene expression profiling of HT-1080 fibrosarcoma cells subjected to CA IX depletion by shRNA silencing. We identified the focal adhesion pathway as being significantly inhibited in the absence of CA IX and confirmed this finding by functional assays. Thus, we obtained the first direct evidence for the role of CA IX in focal adhesion.

View Figures

View References

1	Harris AL: Hypoxia - a key regulatory factor in tumour growth. Nat Rev Cancer. 2:38–47. 2002. View Article : Google Scholar : PubMed/NCBI
2	Pastorekova S, Parkkila S, Pastorek J and Supuran CT: Carbonic anhydrases: current state of the art, therapeutic applications and future prospects. J Enzyme Inhib Med Chem. 19:199–229. 2004.PubMed/NCBI
3	Pastorek J, Pastorekova S, Callebaut I, et al: Cloning and characterization of MN, a human tumor-associated protein with a domain homologous to carbonic anhydrase and a putative helix-loop-helix DNA binding segment. Oncogene. 9:2877–2888. 1994.
4	Wykoff CC, Beasley NJ, Watson PH, et al: Hypoxia-inducible expression of tumor-associated carbonic anhydrases. Cancer Res. 60:7075–7083. 2000.PubMed/NCBI
5	Potter C and Harris AL: Hypoxia inducible carbonic anhydrase IX, marker of tumour hypoxia, survival pathway and therapy target. Cell Cycle. 3:164–167. 2004. View Article : Google Scholar : PubMed/NCBI
6	Svastova E, Hulikova A, Rafajova M, et al: Hypoxia activates the capacity of tumor-associated carbonic anhydrase IX to acidify extracellular pH. FEBS Lett. 577:439–445. 2004. View Article : Google Scholar : PubMed/NCBI
7	Swietach P, Hulikova A, Vaughan-Jones RD and Harris AL: New insights into the physiological role of carbonic anhydrase IX in tumour pH regulation. Oncogene. 29:6509–6521. 2010. View Article : Google Scholar : PubMed/NCBI
8	Raghunand N, Gatenby RA and Gillies RJ: Microenvironmental and cellular consequences of altered blood flow in tumours. Br J Radiol. 76(Spec No 1): S11–S22. 2003. View Article : Google Scholar : PubMed/NCBI
9	Svastova E, Zilka N, Zat’ovicova M, et al: Carbonic anhydrase IX reduces E-cadherin-mediated adhesion of MDCK cells via interaction with beta-catenin. Exp Cell Res. 290:332–345. 2003. View Article : Google Scholar : PubMed/NCBI
10	Svastova E, Witarski W, Csaderova L, et al: Carbonic anhydrase IX interacts with bicarbonate transporters in lamellipodia and increases cell migration via its catalytic domain. J Biol Chem. 287:3392–3402. 2012. View Article : Google Scholar : PubMed/NCBI
11	Takacova M, Holotnakova T, Barathova M, Pastorekova S, Kopacek J and Pastorek J: Src induces expression of carbonic anhydrase IX via hypoxia-inducible factor 1. Oncol Rep. 23:869–874. 2010.PubMed/NCBI
12	Ihnatko R, Kubes M, Takacova M, et al: Extracellular acidosis elevates carbonic anhydrase IX in human glioblastoma cells via transcriptional modulation that does not depend on hypoxia. Int J Oncol. 29:1025–1033. 2006.
13	Kaluz S, Kaluzova M and Stanbridge EJ: Expression of the hypoxia marker carbonic anhydrase IX is critically dependent on SP1 activity. Identification of a novel type of hypoxia-responsive enhancer. Cancer Res. 63:917–922. 2003.PubMed/NCBI
14	Szulc J, Wiznerowicz M, Sauvain MO, Trono D and Aebischer P: A versatile tool for conditional gene expression and knockdown. Nat Methods. 3:109–116. 2006. View Article : Google Scholar : PubMed/NCBI
15	Tarca AL, Draghici S, Khatri P, et al: A novel signaling pathway impact analysis. Bioinformatics. 25:75–82. 2009. View Article : Google Scholar : PubMed/NCBI
16	Pastorekova S, Ratcliffe PJ and Pastorek J: Molecular mechanisms of carbonic anhydrase IX-mediated pH regulation under hypoxia. BJU Int. 101(Suppl 4): 8–15. 2008. View Article : Google Scholar : PubMed/NCBI
17	Chiche J, Ilc K, Laferriere J, et al: Hypoxia-inducible carbonic anhydrase IX and XII promote tumor cell growth by counteracting acidosis through the regulation of the intracellular pH. Cancer Res. 69:358–368. 2009. View Article : Google Scholar : PubMed/NCBI
18	Hulikova A, Zatovicova M, Svastova E, et al: Intact intracellular tail is critical for proper functioning of the tumor-associated, hypoxia-regulated carbonic anhydrase IX. FEBS Lett. 583:3563–3568. 2009. View Article : Google Scholar : PubMed/NCBI
19	Ditte P, Dequiedt F, Svastova E, et al: Phosphorylation of carbonic anhydrase IX controls its ability to mediate extracellular acidification in hypoxic tumors. Cancer Res. 71:7558–7567. 2011. View Article : Google Scholar : PubMed/NCBI
20	Shin HJ, Rho SB, Jung DC, Han IO, Oh ES and Kim JY: Carbonic anhydrase IX (CA9) modulates tumor-associated cell migration and invasion. J Cell Sci. 124:1077–1087. 2011. View Article : Google Scholar : PubMed/NCBI
21	Zamir E and Geiger B: Molecular complexity and dynamics of cell-matrix adhesions. J Cell Sci. 114:3583–3590. 2001.PubMed/NCBI
22	Tu Y, Wu S, Shi X, Chen K and Wu C: Migfilin and Mig-2 link focal adhesions to filamin and the actin cytoskeleton and function in cell shape modulation. Cell. 113:37–47. 2003. View Article : Google Scholar : PubMed/NCBI
23	Sanz-Moreno V, Gadea G, Ahn J, et al: Rac activation and inactivation control plasticity of tumor cell movement. Cell. 135:510–523. 2008. View Article : Google Scholar : PubMed/NCBI
24	Meller N, Irani-Tehrani M, Kiosses WB, Del Pozo MA and Schwartz MA: Zizimin1, a novel Cdc42 activator, reveals a new GEF domain for Rho proteins. Nat Cell Biol. 4:639–647. 2002. View Article : Google Scholar : PubMed/NCBI
25	Cote JF and Vuori K: Identification of an evolutionarily conserved superfamily of DOCK180-related proteins with guanine nucleotide exchange activity. J Cell Sci. 115:4901–4913. 2002. View Article : Google Scholar : PubMed/NCBI
26	Gallagher SM, Castorino JJ and Philp NJ: Interaction of monocarboxylate transporter 4 with beta1-integrin and its role in cell migration. Am J Physiol Cell Physiol. 296:C414–C421. 2009. View Article : Google Scholar : PubMed/NCBI
27	Denker SP and Barber DL: Cell migration requires both ion translocation and cytoskeletal anchoring by the Na-H exchanger NHE1. J Cell Biol. 159:1087–1096. 2002. View Article : Google Scholar : PubMed/NCBI
28	Stock C and Schwab A: Protons make tumor cells move like clockwork. Pflugers Arch. 458:981–992. 2009. View Article : Google Scholar : PubMed/NCBI
29	Jin H and Varner J: Integrins: roles in cancer development and as treatment targets. Br J Cancer. 90:561–565. 2004. View Article : Google Scholar : PubMed/NCBI
30	Nagase H, Visse R and Murphy G: Structure and function of matrix metalloproteinases and TIMPs. Cardiovasc Res. 69:562–573. 2006. View Article : Google Scholar : PubMed/NCBI
31	Rupp PA, Visconti RP, Czirok A, Cheresh DA and Little CD: Matrix metalloproteinase 2-integrin alpha(v)beta3 binding is required for mesenchymal cell invasive activity but not epithelial locomotion: a computational time-lapse study. Mol Biol Cell. 19:5529–5540. 2008. View Article : Google Scholar : PubMed/NCBI
32	Holman CM, Kan CC, Gehring MR and Van Wart HE: Role of His-224 in the anomalous pH dependence of human stromelysin-1. Biochemistry. 38:677–681. 1999. View Article : Google Scholar : PubMed/NCBI
33	Monaco S, Gioia M, Rodriguez J, et al: Modulation of the proteolytic activity of matrix metalloproteinase-2 (gelatinase A) on fibrinogen. Biochem J. 402:503–513. 2007. View Article : Google Scholar : PubMed/NCBI
34	Brooks PC, Stromblad S, Sanders LC, et al: Localization of matrix metalloproteinase MMP-2 to the surface of invasive cells by interaction with integrin alpha v beta 3. Cell. 85:683–693. 1996. View Article : Google Scholar : PubMed/NCBI
35	Hornebeck W, Emonard H, Monboisse JC and Bellon G: Matrix-directed regulation of pericellular proteolysis and tumor progression. Semin Cancer Biol. 12:231–241. 2002. View Article : Google Scholar : PubMed/NCBI
36	Maretzky T, Reiss K, Ludwig A, et al: ADAM10 mediates E-cadherin shedding and regulates epithelial cell-cell adhesion, migration, and beta-catenin translocation. Proc Natl Acad Sci USA. 102:9182–9187. 2005. View Article : Google Scholar : PubMed/NCBI