Skip to main content

Advertisement

Log in

eIF3a is over-expressed in urinary bladder cancer and influences its phenotype independent of translation initiation

  • Published:
Cellular Oncology Aims and scope Submit manuscript

Abstract

Purpose

The eukaryotic translation initiation factor (eIF) 3a, the largest subunit of the eIF3 complex, is a key functional entity in ribosome establishment and translation initiation. In the past, aberrant eIF3a expression has been linked to the pathology of various cancer types but, so far, its expression has not been investigated in transitional cell carcinomas. Here, we investigated the impact of eIF3 expression on urinary bladder cancer (UBC) cell characteristics and UBC patient survival.

Methods and results

eIF3a expression was reduced through inducible knockdown in the UBC-derived cell lines RT112, T24, 5637 and HT1197. As a consequence of eIF3a down-regulation, UBC cell proliferation, clonogenic potential and motility were found to be decreased and, concordantly, UBC tumour cell growth rates were found to be impaired in xenotransplanted mice. Polysomal profiling revealed that reduced eIF3a levels increased the abundance of 80S ribosomes, rather than impairing translation initiation. Microarray-based gene expression and ontology analyses revealed broad effects of eIF3a knockdown on the transcriptome. Analysis of eIF3a expression in primary formalin-fixed paraffin embedded UBC samples of 198 patients revealed that eIF3a up-regulation corresponds to tumour grade and that high eIF3a expression corresponds to longer overall survival rates of patients with low grade tumours.

Conclusions

From our results we conclude that eIF3a expression may have a profound effect on the UBC phenotype and, in addition, may serve as a prognostic marker for low grade UBCs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7

Similar content being viewed by others

References

  1. F. Saletta, Y. Suryo Rahmanto, D.R. Richardson, The translational regulator eIF3a: the tricky eIF3 subunit! Biochim. Biophys. Acta 1806, 275–286 (2010)

    CAS  PubMed  Google Scholar 

  2. Z. Dong, Z. Liu, P. Cui, R. Pincheira, Y. Yang, J. Liu, J.T. Zhang, Role of eIF3a in regulating cell cycle progression. Exp. Cell Res. 315, 1889–1894 (2009)

    Article  CAS  PubMed  Google Scholar 

  3. M. Ploeg, K.K.H. Abden, L.A. Kiemeney, The present and future burden of urinary bladder cancer in the world. World J. Urol. 27, 289–293 (2009)

    Article  PubMed Central  PubMed  Google Scholar 

  4. A. Stenzl, N.C. Cowan, M. DeSantis, G. Jakse, M.A. Kuczyk, A.S. Merseburger, M.J. Ribal, A. Sherif, J.A. Witjes, The updated EAU guidelines on muscle-invasive and metastatic bladder cancer. Eur. Urol. 55, 815–825 (2009)

    Article  PubMed  Google Scholar 

  5. M. Retz, J.E. Gschwend, Medikamentöse Tumortherapie in der Uroonkologie, 2nd edn. (Springer, Berlin, 2010)

    Book  Google Scholar 

  6. G. Johnen, K. Gawrych, H. Bontrup, B. Pesch, D. Taeger, S. Banek, M. Kluckert, H. Wellhäußer, F. Eberle, M. Nasterlack, G. Leng, A. Stenzl, T. Brüning, Performance of survivin mRNA as a biomarker for bladder cancer in the prospective study UroScreen. PLoS One 7, e35363 (2012)

    Article  PubMed Central  PubMed  Google Scholar 

  7. A. Datta, M.E. Adelson, Y. Mogilevkin, E. Mordechai, A.A. Sidi, J.P. Trama, Oncoprotein DEK as a tissue and urinary biomarker for bladder cancer. BMC Cancer 11, 234 (2011)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  8. V. Cebrian, M. Fierro, E. Orenes-Piñero, L. Grau, P. Moya, T. Ecke, M. Alvarez, M. Gil, F. Algaba, J. Bellmunt, C. Cordon-Cardo, J. Catto, A. López-Beltrán, M. Sánchez-Carbayo, KISS1 methylation and expression as tumor stratification biomarkers and clinical outcome prognosticators for bladder cancer patients. Am. J. Pathol. 179, 540–546 (2011)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  9. L.G. Koss, E.M. Tiamson, M.A. Robbins, Mapping cancerous and precancerous bladder changes. JAMA 227, 281–286 (1974)

    Article  CAS  PubMed  Google Scholar 

  10. P.E. Spiess, B. Czerniak, Dual-track pathway of bladder carcinogenesis: practical implications. Arch. Pathol. Lab. Med. 130, 844–852 (2006)

    CAS  PubMed  Google Scholar 

  11. T.V. Pestova, J.R. Lorsch, C.U.T. Hellen, The Mechanism of Translation Initiation in Eukaryotes, in Translational Control in Biology and Medicine, ed. by M.B. Mathews, N. Sonenberg, J.W.B. Hershey (Cold Spring Harbor Laboratory Press, New York, 2007), pp. 87–128

    Google Scholar 

  12. A. Unbehaun, S.I. Borukhov, C.U.T. Hellen, T.V. Pestova, Release of initiation factors from 48S complexes during ribosomal subunit joining and the link between establishment of codon-anticodon base-pairing and hydrolysis of eIF2-bound GTP. Genes Dev. 18, 3078–3093 (2004)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  13. T. Kouba, I. Danyi, S. Gunisova, V. Munzarova, V. Vlckova, L. Cuchalova, A. Neueder, P. Milkereit, L.S. Valasek, Small ribosomal protein RPS0 stimulates translation initiation by mediating 40S-binding of eIF3 via its direct contact with the eIF3a/TIF32 subunit. PLoS One 7, e40464 (2012)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  14. R.Y. Liu, Z. Dong, J. Liu, J.Y. Yin, L. Zhou, X. Wu, Y. Yang, W. Mo, W. Huang, S.K. Khoo, J. Chen, D. Petillo, B.T. Teh, C.N. Qian, J.T. Zhang, Role of eIF3a in regulating cisplatin sensitivity and nucleotide excision repair of nasopharyngeal carcinomas. Oncogene 30, 4814–4823 (2012)

    Article  Google Scholar 

  15. J.Y. Yin, J. Shen, Z.Z. Dong, Q. Huang, M.Z. Zhong, D.Y. Feng, H.H. Zhou, J.T. Zhang, Z.Q. Liu, Effect of eIF3a on response of lung cancer patients to platinum-based chemotherapy by regulating DNA repair. Clin. Cancer Res. 17, 4600–4609 (2011)

    Article  CAS  PubMed  Google Scholar 

  16. R. Spilka, K. Laimer, F. Bachmann, G. Spizzo, A. Vogetseder, M. Wieser, H. Müller, J. Haybaeck, P. Obrist, Overexpression of eIF3a in squamous cell carcinoma of the oral cavity and its putative relation to chemotherapy response. J. Oncol. 901956 (2012)

  17. R. Gentleman, V. Carey, D. Bates, B. Bolstad, Bioconductor: open software development for computational biology and bioinformatics. Genome Biol. 5, R80 (2004)

    Article  PubMed Central  PubMed  Google Scholar 

  18. Z. Wu, R.A. Irizarry, R. Gentleman, F. Martinez Murillo, F. Spencer, A model based background adjustment for oligonucleotide expression arrays. Johns Hopkins University, Dept. of Biostatistics Working Papers. Working Paper 1 (2004)

  19. G.K. Smyth, Linear models and empirical Bayes methods for assessing differential expression in microarray experiments. Stat. Appl. Genet. Mol. Biol. 3, e29 (2004)

    Google Scholar 

  20. Y. Benjamini, Y. Hochberg, Controlling the false discovery rate: a practical and powerful approach to multiple testing. J. R Stat. Soc. Ser. B 57, 289–300 (1995)

    Google Scholar 

  21. S. Falcon, R. Gentleman, Using GOstats to test gene lists for GO term association. Bioinformatics 23, 257–258 (2007)

    Article  CAS  PubMed  Google Scholar 

  22. G. Spizzo, D. Fong, M. Wurm, C. Ensinger, P. Obrist, C. Hofer, G. Mazzoleni, G. Gastl, P. Went, EpCAM expression in primary tumour tissues and metastases: an immunohistochemical analysis. J. Clin. Pathol. 64, 415–420 (2011)

    Article  PubMed Central  PubMed  Google Scholar 

  23. J. Bubeník, M. Baresová, V. Viklický, J. Jakoubková, H. Sainerová, J. Donner, Established cell line of urinary bladder carcinoma (T24) containing tumour-specific antigen. Int. J. Cancer 11, 765–773 (1973)

    Article  PubMed  Google Scholar 

  24. J. Fogh, Cultivation, characterization and identification of human tumor cells with emphasis on kidney, testis and bladder tumors. Natl. Cancer Inst. Monogr. 49, 5–9 (1978)

    PubMed  Google Scholar 

  25. C.J. Marshall, L.M. Franks, A.W. Carbonell, Markers of neoplastic transformation in epithelial cell lines derived from human carcinomas. J. Nat. Cancer Inst. 58, 1743–1751 (1977)

    CAS  PubMed  Google Scholar 

  26. V. Croons, W. Martinet, A.G. Herman, G.R. De Meyer, Differential effect of the protein synthesis inhibitors puromycin and cycloheximide on vascular smooth muscle cell viability. J. Pharmacol. Exp. Ther. 325, 824–832 (2008)

    Article  CAS  PubMed  Google Scholar 

  27. S. Pestka, Inhibitors of ribosome functions. Annu. Rev. Microbiol. 25, 487–562 (1971)

    Article  CAS  PubMed  Google Scholar 

  28. X. Wang, C.G. Proud, The mTOR pathway in the control of protein synthesis. Physiology 21, 362–369 (2006)

    Article  CAS  PubMed  Google Scholar 

  29. K. Inoki, H. Ouyang, Y. Li, K.L. Guan, Signaling by target of rapamycin proteins in cell growth control. Microbiol. Mol. Biol. Rev. 69, 79–100 (2005)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  30. S. Khoshnevis, S. Gunisová, V. Vlcková, T. Kouba, P. Neumann, P. Beznosková, R. Ficner, L.S. Valásek, Structural integrity of the PCI domain of eIF3a/TIF32 is required for mRNA recruitment to the 43S pre-initiation complexes. Nucleic Acids Res. 42, 4123–4139 (2014)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  31. A.G. Hinnebusch, eIF3: a versatile scaffold for translation initiation complexes. Trends Biochem. Sci. 31, 553–562 (2006)

    Article  CAS  PubMed  Google Scholar 

  32. F. Bachman, R. Baenziger, M.M. Burger, Cloning of a novel protein overexpressed in human mammary carcinoma. Cancer Res. 57, 988–994 (1997)

    Google Scholar 

  33. A. Dellas, J. Torhorst, F. Bachmann, R. Baenziger, E. Schultheiss, M.M. Burger, Expression of p150 in cervical neoplasia and its potential value in predicting survival. Cancer 83, 1376–1383 (1998)

    Article  CAS  PubMed  Google Scholar 

  34. G. Chen, M.M. Burger, P150 expression and its prognostic value in squamous cell carcinoma of the esophagus. Int. J. Cancer 84, 95–100 (1999)

    Article  CAS  PubMed  Google Scholar 

  35. R. Pincheira, Q. Chen, J.T. Zhang, Identification of a 170-kDa protein over-expressed in lung cancers. Br. J. Cancer 84, 1520–1527 (2001)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  36. G. Chen, M.M. Burger, P150 Overexpression in gastric carcinoma: the association with p53, apoptosis and cell proliferation. Int. J. Cancer 112, 393–398 (2004)

    Article  CAS  PubMed  Google Scholar 

  37. J. Haybaeck, T. O’Connor, R. Spilka, G. Spizzo, C. Ensinger, G. Mikuz, T. Brunhuber, A. Vogetseder, I. Theurl, W. Salvenmoser, H. Draxl, R. Baenziger, F. Bachmann, G. Schaefer, M.M. Burger, P. Obrist, Overexpression of p150, a part of the large subunit of the eukaryotic translation initiation factor 3, in colon cancer. Anticancer Res. 30, 1047–1056 (2010)

    CAS  PubMed  Google Scholar 

  38. K.C. Halling, W. King, I.A. Sokolova, R.J. Karnes, R.G. Meyer, E.L. Powell, T.J. Sebo, J.C. Cheville, A.C. Clayton, K.L. Krajnik, T.A. Ebert, R.E. Nelson, H.M. Burkhardt, S. Ramakumar, C.S. Stewart, V.S. Pankratz, M.M. Lieber, M.L. Blute, H. Zincke, S.A. Seelig, R.B. Jenkins, D.J. O’Kane, A comparison of BTA stat, hemoglobin dipstick, telomerase and Vysis UroVysion assays for the detection of urothelial carcinoma in urine. J. Urol. 167, 2001–2006 (2002)

    Article  CAS  PubMed  Google Scholar 

  39. H. Jamshidian, K. Kor, M. Djalali, Urine concentration of nuclear matrix protein 22 for diagnosis of transitional cell carcinoma of bladder. Urol. J. 5, 243–247 (2008)

    PubMed  Google Scholar 

  40. L.P. Huang, D. Savoly, A.A. Sidi, M.E. Adelson, E. Mordechai, J.P. Trama, CIP2A protein expression in high-grade, high-stage bladder cancer. Cancer Med. 1, 76–81 (2012)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  41. Y.C. Lu, C.N. Chen, B. Wang, W.M. Hsu, S.T. Chen, K.J. Chang, C.C. Chang, H. Lee, Changes in tumor growth and metastatic capacities of J82 human bladder cancer cells suppressed by down-regulation of calreticulin expression. Am. J. Pathol. 179, 1425–1433 (2011)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  42. M. Unoki, J.D. Kelly, D.E. Neal, B.A. Ponder, Y. Nakamura, R. Hamamoto, UHRF1 is a novel molecular marker for diagnosis and the prognosis of bladder cancer. Br. J. Cancer 101, 98–105 (2009)

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  43. D. Silvera, S.C. Formenti, R.J. Schneider, Translational control in cancer. Nat. Rev. Cancer 10, 254–266 (2010)

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

We thank Theresa Eder, Veronika Rauch, Gertrude Zisser and Isolde Gunsch for their excellent technical assistance. We thank Mag. Karin Osibow for critical reading of our manuscript.

Conflict of interest

The authors declare that there is no conflicts of interest.

Author information

Authors and Affiliations

Authors

Corresponding authors

Correspondence to Rita Spilka or Johannes Haybaeck.

Additional information

Rita Spilka, Christina Ernst, Peter Obrist and Johannes Haybaeck contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplemental Fig. 1

Heat map displaying the gene ontology analysis results for ‘cell proliferation’ genes sorted by expression (GIF 902 kb)

High resolution image (TIFF 337 kb)

Supplemental Table 1

(PDF 3166 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Spilka, R., Ernst, C., Bergler, H. et al. eIF3a is over-expressed in urinary bladder cancer and influences its phenotype independent of translation initiation. Cell Oncol. 37, 253–267 (2014). https://doi.org/10.1007/s13402-014-0181-9

Download citation

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13402-014-0181-9

Keywords

Navigation