Molecular Markers of Kidney Cancer Progression, Association with Efficiency of Pazopanib Therapy

Liudmila V. Spirina¹, Evgeny A. Usynin², Irina V. Kondakova², Zahar A. Yurmazov², Elena M. Slonimskaya¹

¹Tomsk Cancer Research Institute, Siberian State Medical University, Tomsk, Russia.
²Tomsk Cancer Research Institute, Tomsk, Russia.

DOI: 10.4236/jbise.2015.811072                

Abstract

Purpose: The aim of the study is to reveal associations between NF-κB, HIF-1alpha, VEGF expres-sions, proteasome and calpain activities with tumor progression in patients with kidney cancers and to find molecular parameters, associated with the effective pazopanib therapy. 93 patients with clear cell kidney cancers are included in investigation. 26 patients with disseminated kidney cancer have the pazopanib therapy. Methods: Transcription factors, VEGF, VEGFR2 and p-m-TOR expression are measured by ELISA kits. Proteasome and calpain activity are determined using specific fluorogenic substrate. Results: It is found the increase of NF-κB, HIF-1 expression in cancer tissues followed the hematogenic metastasis development. Coefficient NF-κB р65/р50 and VEGF expression are increased in cancer tissues with single metastasis and are decreased in cancer tissues with multiple ones. It is observed in the low proteasome activity in metastatic cancer tissues. The partial cancer regression is revealed in 29.6% of patients treated with pazopanib, cancer stabilization—in 61.5% of patients and cancer progression—in 11.5% of patients. The increased level of transcription factors NF-κB, HIF-1, growth factor VEGF and high proteasome activity in cancer tissues before targeted therapy are associated with the effective treatment. It is obtained the significant decrease of investigated markers after pazopanib application in metastatic kidney cancer patients. Conclusion: Coefficient NF-κB р65/р50, VEGF expression and proteases activities are the potential prognostic molecular markers of hematogenic metastasis development in kidney cancers. NF-κB, HIF-1 and VEGF levels can be considered as additional molecular markers predicting the effective pazopanib therapy.

Keywords

Kidney Cancer, NF-κB, HIF-1, VEGF, VEGFR2, m-TOR, Proteasome, Calpain, Pazopanib

Conflicts of Interest

The authors declare no conflicts of interest.

References

[1]	Kidney Cancer (2014) Cancer.Net Editorial Board. http://www.cancer.net/cancer-types/kidney-cancer/statistics
[2]	Kidney Cancer (2014) National Cancer Institute at the National Institutes of Health. http://www.cancer.gov/cancertopics/types/kidney
[3]	Keefe, S.M., Nathanson, K.L. and Rathmell, W.K. (2013) The Molecular Biology of Renal Cell Carcinoma. Seminars in Oncology, 40, 421-428. http://dx.doi.org/10.1053/j.seminoncol.2013.05.006
[4]	Na, X., Wu, G., Ryan, C.K., Schoeb, S.R., di’Santagnese, P.A. and Messing, E.M. (2003) Overproduction of Vascular Endothelial Growth Factor Related to von Hippel-Lindau Tumor Suppressor Gene Mutations and Hypoxia-Inducible Factor-1α Expression in Renal Cell Carcinomas. Journal of Urology, 170, 588-592. http://dx.doi.org/10.1097/01.ju.0000074870.54671.98
[5]	Hoffmann, A. and Baltimore, D. (2006) Circuitry of Nuclear Factor κB Signaling. Immunological Reviews, 210, 171-186.
[6]	Zhou, J., Kohl, R., Herr, B., Frank, R. and Brune, B. (2006) Calpain Mediates a von Hippel-Lindau Protein-Independent Destruction of Hypoxia-Inducible Factor-1α. Molecular Biology of the Cell, 17, 1549-558. http://dx.doi.org/10.1091/mbc.E05-08-0770
[7]	Klatte, T., Seligson, D.B., Riggs, S.B., Leppert, J.T. and Berkman, M.K. (2007) Hypoxia-Inducible Factor-1α in Clear Cell Renal Cell Carcinoma. Clinical Cancer Research, 13, 7388-7393. http://dx.doi.org/10.1158/1078-0432.CCR-07-0411
[8]	Goldberg, A.L. (2007) Functions of the Proteasome: From Protein Degradation and Immune Surveillance to Cancer Therapy. Biochemical Society Transactions, 35, 12-17. http://dx.doi.org/10.1042/BST0350012
[9]	Kostadinova, R.M., Nawrocki, A.R., Frey, F.J. and Frey, B.M. (2005) Tumor Necrosis Factor α and Phorbol 12-Myri- state-13-acetate Down-Regulate Human 11β-Hydroxysteroid Dehydrogenase Type 2 through p50/p50 NF-κΒ Homodimers and Egr-1. FASEB Journal, 19, 650-652.
[10]	Marui, N., Medford, R.M. and Ahmad, M. (2005) Activation of RelA Homodimers by Tumor Necrosis Factor α: A Possible Transcriptional Activator in Human Vascular Endothelial Cells. Biochemical Journal, 390, 317-324. http://dx.doi.org/10.1042/BJ20041659
[11]	Juvekar, A.S., Manna, S., Ramaswami, S., Chang, T.P., Vu, H.Y., Ghosh, C.C., Celiker, M.Y. and Vancurova, I. (2011) Bortezomib Induces Nuclear Translocation of IkBα Resulting in Gene-Specific Suppression of NF-κΒ-Dependent Transcription and Induction of Apoptosis in CTCL. Molecular Cancer Research, 9, 183-194. http://dx.doi.org/10.1158/1541-7786.MCR-10-0368
[12]	Conner, J.R., Smirnova, I.I., Moseman, A.P. and Poltorak, A. (2010) IRAK1BP1 Inhibits Inflammation by Promoting Nuclear Translocation of NF-κΒp50. Proceedings of the National Academy of Sciences of the United States of America, 107, 11477-11482. http://dx.doi.org/10.1073/pnas.1006894107
[13]	van Uden, P., Kenneth, N.S. and Rocha, S. (2008) Regulation of Hypoxia-Inducible Factor-1α by NF-κΒ. Biochemical Journal, 412, 477-484.
[14]	Baldwin, A.S. (1996) The NF-κΒ and IκΒ Proteins: New Discoveries and Insights. Annual Review of Immunology, 14, 649-683. http://dx.doi.org/10.1146/annurev.immunol.14.1.649
[15]	Kelvin, J.A.D. and Reshma, S. (2006) Preferential Degradation of Oxidized Proteins by the 20S Proteasome May Be Inhibited in Aging and in Inflammatory Neuromuscular Diseases. Neurology, 66, 93-96. http://dx.doi.org/10.1212/01.wnl.0000192308.43151.63
[16]	Sorokin, A.V., Kim, E.R. and Ovchinnikov, L.P. (2009) Proteasome System of Protein Degradation and Processing. Biochemistry (Moscow), 74, 1411-1442. http://dx.doi.org/10.1134/S000629790913001X
[17]	Chen, C., Seth, A.K. and Aplin, A.E. (2006) Genetic and Expression Aberrations of E3 Ubiquitin Ligases in Human Breast Cancer. Molecular Cancer Research, 4, 695-707. http://dx.doi.org/10.1158/1541-7786.MCR-06-0182
[18]	Voutsadakis, I.A. (2007) Pathogenesis of Colorectal Carci-noma and Therapeutic Implications: The Role of the Ubiquitin-Proteasome System and Cox-2. Journal of Cellular and Molecular Medicine, 11, 252-337. http://dx.doi.org/10.1111/j.1582-4934.2007.00032.x
[19]	An, J. and Rettig, M.B. (2005) Mechanism of Von Hippel-Lindau Protein-Mediated Suppression of Nuclear Factor Kappa B Activity. Molecular and Cellular Biology, 25, 7546-7556. http://dx.doi.org/10.1128/MCB.25.17.7546-7556.2005
[20]	An, J. and Rettig, M.B. (2007) Epidermal Growth Factor Receptor Inhibition Sensitizes Renal Cell Carcinoma Cells to the Cytotoxic Effects of Bortezomib. Molecular Cancer Therapeutics, 6, 61-69. http://dx.doi.org/10.1158/1535-7163.MCT-06-0255
[21]	Spirina, L.V., Kondakova, I.V., Usynin, Y.A. and Vintizenko S.I. (2008) Angiogenesis Regulation in Renal and Bladder Cancers. Siberian Journal of oncology, 4, 65-70.
[22]	Wu, W.K., Volta, V., Cho, C.H., Wu, Y.C., Li, H.T., Yu, L., Li, Z.J. and Sung, J.J. (2009) Repression of Protein Translation and mTOR Signaling by Proteasome Inhibitor in Colon Cancer Cells. Biochemical and Biophysical Research Communications, 386, 598-601. http://dx.doi.org/10.1016/j.bbrc.2009.06.080
[23]	Li, C., Chen, S., Yue, P., Deng, X., Lonial, S., Khuri, F.R. and Sun, S.Y. (2010) Proteasome Inhibitor PS-341 (Bortezomib) Induces Calpain-Dependent IκBα Degradation. Journal of Biological Chemistry, 285, 16096-16104. http://dx.doi.org/10.1074/jbc.M109.072694
[24]	Moorty, A.K., Savinova, O.V., Ho, J.Q., Wang, V.Y., Vu, D. and Ghosh, G. (2006) The 20S Proteasome Processes NF-κΒ1 p105 into p50 in a Translation-Independent Manner. The EMBO Journal, 25, 1945-1956. http://dx.doi.org/10.1038/sj.emboj.7601081
[25]	Sorimachi, H., Hata, S. and Ono, Y. (2011) Calpain Chronicle an Enzyme Family under Multidisciplinary Characterization. Proceedings of the Japan Academy, Series B, 87, 287-327. http://dx.doi.org/10.2183/pjab.87.287
[26]	Smith, I.J. and Dodd, S.L. (2007) Calpain Activation Causes a Proteasome Dependent Increase in Protein Degradation and Inhibits the Akt Signaling Pathway in Rat Diaphragm Muscle. Experimental Physiology, 92, 561-573. http://dx.doi.org/10.1113/expphysiol.2006.035790
[27]	Yagasaki, H., Kawata, N., Takimoto, Y. and Nemoto, N. (2003) Histopathological Analysis of Angiogenic Factors in Renal Cell Carcinoma. International Journal of Urology, 10, 220-227.
[28]	Spirina, L.V., Kondakova, I.V., Usynin, E.A. and Yurmazov, Z.A. (2012) Expression Regulation of Transcription Factors and Endothelial Growth Factor by Proteosomal System in Patients with Metastatic Renal Carcinoma. Journal of N. N. Blokhin Russian Cancer Research Center, 23, 27-32.
[29]	Lidgren, A., Hedberg, Y., Grankvist, K., Rasmuson, T., Vasko, J. and Ljungberg, B. (2005) The Expression of Hypoxia-Inducible Factor 1α Is a Favorable Independent Prognostic Factor in Renal Cell Carcinoma. Clinical Cancer Research, 11, 1129-1135.
[30]	Morais, C., Gobe, G., Johnson, D.W. and Healy, H. (2011) The Emerging Role of Nuclear Factor Kappa B in Renal Cell Carcinoma. The International Journal of Biochemistry & Cell Biology, 43, 1537-1549. http://dx.doi.org/10.1016/j.biocel.2011.08.003
[31]	Atkins, M., Regan, M. and McDermott, D. (2004) Update on the Role of Interleukin 2 and Other Cytokines in the Treatment of Patients with Stage IV Renal Carcinoma. Clinical Cancer Research, 10, 6342S-6346S. http://dx.doi.org/10.1158/1078-0432.ccr-040029
[32]	Guerin, M., Salem, N., Walz, J., Dermeche, S. and Gravis, G. (2013) Major Response with Sorafenib in Advanced Renal Cell Carcinoma after 14 Years of Follow-Up. World Journal of Surgical Oncology, 11, 243-247. http://dx.doi.org/10.1186/1477-7819-11-243
[33]	Levy, A., Hollebecque, A., Ferte, C., Koscielny, S., Fernandez, M., Soria, J.-C. and Massard, C. (2013) Tumor Assessment Criteria in Phase I Trials: Beyond RECIST. Journal of Clinical Oncology, 31, 395. http://dx.doi.org/10.1200/JCO.2012.46.2184
[34]	Hugonnet, F., Fournier, L., Medioni, J., Smadja, C., Hindie, E., Huchet, V., Itti, E., Cuenod, C.-A., Chatellier, G., Oudard, S. and Faraggi, M. (2011) Metastatic Renal Cell Carcinoma: Relationship between Initial Metastasis Hypoxia, Change after 1 Month’s Sunitinib, and Therapeutic Response: An 18F-Fluoromisonidazole PET/CT Study. Journal of Nuclear Medicine, 52, 1048-1055. http://dx.doi.org/10.2967/jnumed.110.084517
[35]	Dornbusch, J., Zacharis, A., Meinhardt, M., Erdmann, K. and Wolff, I. (2013) Analyses of Potential Predictive Markers and Survival Data for a Response to Sunitinib in Patients with Metastatic Renal Cell Carcinoma. PLoS ONE, 8, e76386. http://dx.doi.org/10.1371/journal.pone.0076386
[36]	Rosa, R., Damiano, V., Nappi, L., Formisano, L., Massari, F., Scarpa, A., Martignoni, G., Bianco, R. and Tortora, G. (2013) Angiogenic and Signalling Proteins Correlate with Sensitivity to Sequential Treatment in Renal Cell Cancer. British Journal of Cancer, 109, 686-693. http://dx.doi.org/10.1038/bjc.2013.360
[37]	Xu, C.V., Bing, N.X., Ball, H.A., Rajagopalan, D., Sternberg, C.N., Hutson, T.E., de Souza, P., Xue, Z.G., McCann, L., King, K.S., Ragone, L.J., Whittaker, J.C., Spraggs, C.F., Cardon, L.R., Mooser, V.E. and Pandite, L.N. J. (2011) Pazopanib Efficacy in Renal Cell Carcinoma: Evidence for Predictive Genetic Markers in Angiogenesis-Related and Exposure-Related Genes. Journal of Clinical Oncology, 29, 2557-2564. http://dx.doi.org/10.1200/JCO.2010.32.9110
[38]	Abramova, E.B., Astakhova, T.M., Erokhov, P.A. and Sharova, N.P. (2004) Multiple Forms of the Proteasomes and Some Approaches to Their Separation. Izvestiya Akademii Nauk—Seriya Bi-ologicheskaya, 2, 150-156.
[39]	Ben-Shahar, S., Komlosh, A., Nadav, E., Shaked, I., Ziv, T., Admon, A., Martino, G.N. and Ress, Y. (1999) 26S Proteasome-Mediated Production of an Authentic Major Histocompatibility Class I-Restricted Epitope from an Intact Protein Substrate. Journal of Biological Chemistry, 274, 21963-21972. http://dx.doi.org/10.1074/jbc.274.31.21963
[40]	Sandmann, S., Prenzel, F., Shaw, L., Schauer, R. and Unger, T. (2002) Activity Profile of Calpains I and II in Chronically Infracted Rat Myocardium—Influence of the Calpain Inhibitor CAL 9961. British Journal of Pharmacology, 135, 1951-1958. http://dx.doi.org/10.1038/sj.bjp.0704661
[41]	Sato, K., Tsuchiya, N., Sasaki, R., Shimoda, N., Satoh, S., Ogawa, O. and Kato, T. (1999) Increased Serum Levels of Vascular Endothelial Growth Factor in Patients with Renal Cell Carcinoma. Japanese Journal of Cancer Research, 90, 874-879. http://dx.doi.org/10.1111/j.1349-7006.1999.tb00829.x
[42]	Glading, A., Reynolds, I.J. and Shiraha, H. (2004) Epidermal Growth Factor Activates M-Calpain (Calpain II), at Least in Part, by Extracellular Signal-Regulated Kinase-Mediated Phosphorylation. Molecular and Cellular Biology, 24, 2499-2512. http://dx.doi.org/10.1128/MCB.24.6.2499-2512.2004