Abstract
Renal cell carcinoma (RCC) is the third most frequent malignancy within urological oncology. Sunitinib has been used as the standard of treatment for first-line RCC therapy. Understanding mechanisms of sunitinib resistance in RCC cell is important for clinical therapy and drug development. We established sunitinib resistant RCC cells by treating cells with increasing concentrations of sunitinib and named resistant cells as RCC/SR. Lefty A, an important embryonic morphogen, was increased in RCC/SR cells. Targeted inhibition of Lefty via its siRNAs restored the sensitivity of renal resistant cells to sunitinib treatment. It was due to that si-Lefty can decrease the expression of interleukin-8 (IL-8) in RCC/SR cells. Knockdown of IL-8 abolished Lefty-regulated sunitinib sensitivity of RCC cells. Mechanistically, Lefty can regulate IL-8 transcription via activation of p65, one major transcription factor of IL-8. Collectively, our present revealed that Lefty A can regulate sunitinib sensitivity of RCC cells of via NF-κB/IL-8 signals. It indicated that targeted inhibition of Lefty might be a potent approach to overcome sunitinib resistance of RCC.
-
Author contributions: All the authors have accepted responsibility for the entire content of this submitted manuscript and approved submission.
-
Research funding: None declared.
-
Conflict of interest statement: The authors declare no conflict of interest.
-
Consent for publication: All authors give the consent for the publish of this study.
-
Availability of data and material: All data and material are available.
References
Achermann, C., Stenner, F., and Rothschild, S.I. (2016). Treatment, outcome and prognostic factors in renal cell carcinoma - a single center study (2000-2010). J. Cancer 7: 921–927, https://doi.org/10.7150/jca.15228.Search in Google Scholar
Akiya, M., Yamazaki, M., Matsumoto, T., Kawashima, Y., Oguri, Y., Kajita, S., Kijima, D., Chiba, R., Yokoi, A., Takahashi, H., et al.. (2017). Identification of LEFTY as a molecular marker for ovarian clear cell carcinoma. Oncotarget 8: 63646–63664, https://doi.org/10.18632/oncotarget.18882.Search in Google Scholar
Bi, L.K., Zhou, N., Liu, C., Lu, F.D., Lin, T.X., Xuan, X.J., Jiang, C., Han, J.L., Huang, H., Zhang, C.X., et al.. (2014). Kidney cancer cells secrete IL-8 to activate Akt and promote migration of mesenchymal stem cells. Urol. Oncol.: Semin. Original 32: 607–612, https://doi.org/10.1016/j.urolonc.2013.10.018.Search in Google Scholar
Brunen, D., Willems, S.M., Kellner, U., Midgley, R., Simon, I., and Bernards, R. (2013). TGF-beta an emerging player in drug resistance. Cell Cycle 12: 2960–2968, https://doi.org/10.4161/cc.26034.Search in Google Scholar
Cavallari, C., Fonsato, V., Herrera, M.B., Bruno, S., Tetta, C., and Camussi, G. (2013). Role of Lefty in the anti tumor activity of human adult liver stem cells. Oncogene 32: 819–826, https://doi.org/10.1038/onc.2012.114.Search in Google Scholar
Chen, R., Halder, G., Zhang, Z., and Mardon, G. (1999). Signaling by the TGF-beta homolog decapentaplegic functions reiteratively within the network of genes controlling retinal cell fate determination in Drosophila. Development 126: 935–943, https://doi.org/10.1242/dev.126.5.935.Search in Google Scholar
Cheng, S.K., Olale, F., Brivanlou, A.H., and Schier, A.F. (2004). Lefty blocks a subset of TGFbeta signals by antagonizing EGF-CFC coreceptors. PLoS Biol. 2: E30, https://doi.org/10.1371/journal.pbio.0020030.Search in Google Scholar
Corro, C., Healy, M.E., Engler, S., Bodenmiller, B., Li, Z., Schraml, P., Weber, A., Frew, I.J., Rechsteiner, M., and Moch, H. (2019). IL-8 and CXCR1 expression is associated with cancer stem cell-like properties of clear cell renal cancer. J. Pathol. 248: 377–389, https://doi.org/10.1002/path.5267.Search in Google Scholar
Dai, C.L., Liang, Y.J., Wang, Y.S., Tiwari, A.K., Yan, Y.Y., Wang, F., Chen, Z.S., Tong, X.Z., and Fu, L.W. (2009). Sensitization of ABCG2-overexpressing cells to conventional chemotherapeutic agent by sunitinib was associated with inhibiting the function of ABCG2. Cancer Lett. 279: 74–83, https://doi.org/10.1016/j.canlet.2009.01.027.Search in Google Scholar
Derynck, R., Zhang, Y., and Feng, X.H. (1998). Smads: transcriptional activators of TGF-β responses. Cell 95: 737–740, https://doi.org/10.1016/s0092-8674(00)81696-7.Search in Google Scholar
Dvash, T., Sharon, N., Yanuka, O., and Benvenisty, N. (2007). Molecular analysis of LEFTY-expressing cells in early human embryoid bodies. Stem Cell. 25: 465–472, https://doi.org/10.1634/stemcells.2006-0179.Search in Google Scholar
Esteban, M.A., Bao, X., Zhuang, Q., Zhou, T., Qin, B., and Pei, D. (2012). The mesenchymal-to-epithelial transition in somatic cell reprogramming. Curr. Opin. Genet. Dev. 22: 423–428, https://doi.org/10.1016/j.gde.2012.09.004.Search in Google Scholar
Hamada, H., Meno, C., Watanabe, D., and Saijoh, Y. (2002). Establishment of vertebrate left-right asymmetry. Nat. Rev. Genet. 3: 103–113, https://doi.org/10.1038/nrg732.Search in Google Scholar
Huang, D., Ding, Y., Li, Y., Luo, W.M., Zhang, Z.F., Snider, J., Vandenbeldt, K., Qian, C.N., and Teh, B.T. (2010). Sunitinib acts primarily on tumor endothelium rather than tumor cells to inhibit the growth of renal cell carcinoma. Cancer Res 70: 1053–1062, https://doi.org/10.1158/0008-5472.can-09-3722.Search in Google Scholar
Joosten, S.C., Hamming, L., Soetekouw, P.M., Aarts, M.J., Veeck, J., van Engeland, M., and Tjan-Heijnen, V.C. (2015). Resistance to sunitinib in renal cell carcinoma: from molecular mechanisms to predictive markers and future perspectives. Biochim. Biophys. Acta 1855: 1–16, https://doi.org/10.1016/j.bbcan.2014.11.002.Search in Google Scholar
Jundi, K. and Greene, C.M. (2015). Transcription of Interleukin-8: how altered regulation can affect cystic fibrosis lung disease. Biomolecules 5: 1386–1398, https://doi.org/10.3390/biom5031386.Search in Google Scholar
Liu, Y.N., Chang, T.H., Tsai, M.F., Wu, S.G., Tsai, T.H., Chen, H.Y., Yu, S.L., Yang, J.C.H., and Shih, J.Y. (2015). IL-8 confers resistance to EGFR inhibitors by inducing stem cell properties in lung cancer. Oncotarget 6: 10415–10431, https://doi.org/10.18632/oncotarget.3389.Search in Google Scholar
Ma, J., Ren, Z., Ma, Y., Xu, L., Zhao, Y., Zheng, C., Fang, Y., Xue, T., Sun, B., and Xiao, W. (2009). Targeted knockdown of EGR-1 inhibits IL-8 production and IL-8-mediated invasion of prostate cancer cells through suppressing EGR-1/NF-κB synergy. J. Biol. Chem. 284: 34600–34606, https://doi.org/10.1074/jbc.m109.016246.Search in Google Scholar
Markowitsch, S.D., Schupp, P., Lauckner, J., Vakhrusheva, O., Slade, K.S., Mager, R., Efferth, T., Haferkamp, A., and Juengel, E. (2020). Artesunate inhibits growth of sunitinib-resistant renal cell carcinoma cells through cell cycle arrest and induction of ferroptosis. Cancers 12, https://doi.org/10.3390/cancers12113150.Search in Google Scholar
Matsumoto, T., Chino, H., Akiya, M., Hashimura, M., Yokoi, A., Tochimoto, M., Nakagawa, M., Jiang, Z., and Saegusa, M. (2020). Requirements of LEFTY and Nodal overexpression for tumor cell survival under hypoxia in glioblastoma. Mol. Carcinog. 59: 1409–1419, https://doi.org/10.1002/mc.23265.Search in Google Scholar
Matsumoto, T., Yokoi, A., Hashimura, M., Oguri, Y., Akiya, M., and Saegusa, M. (2018). TGF--mediated LEFTY/Akt/GSK-3/Snail axis modulates epithelial-mesenchymal transition and cancer stem cell properties in ovarian clear cell carcinomas. Mol. Carcinog. 57: 957–967, https://doi.org/10.1002/mc.22816.Search in Google Scholar
Meno, C., Ito, Y., Saijoh, Y., Matsuda, Y., Tashiro, K., Kuhara, S., and Hamada, H. (1997). Two closely-related left-right asymmetrically expressed genes, Lefty-1 and Lefty-2: their distinct expression domains, chromosomal linkage and direct neuralizing activity in Xenopus embryos. Gene Cell 2: 513–524, https://doi.org/10.1046/j.1365-2443.1997.1400338.x.Search in Google Scholar
Oshimori, N., Oristian, D., and Fuchs, E. (2015). TGF-β promotes heterogeneity and drug resistance in squamous cell carcinoma. Cell 160: 963–976, https://doi.org/10.1016/j.cell.2015.01.043.Search in Google Scholar
Peng, Q., Wang, L., Zhao, D., Lv, Y., Wang, H., Chen, G., Wang, J., and Xu, W. (2019). Overexpression of FZD1 is associated with a good prognosis and resistance of sunitinib in clear cell renal cell carcinoma. J. Cancer 10: 1237–1251, https://doi.org/10.7150/jca.28662.Search in Google Scholar
Prud'homme, G.J. (2012). Cancer stem cells and novel targets for antitumor strategies. Curr. Pharmaceut. Des. 18: 2838–2849, https://doi.org/10.2174/138161212800626120.Search in Google Scholar
Rossi, S.H., Klatte, T., Usher-Smith, J., and Stewart, G.D. (2018). Epidemiology and screening for renal cancer. World J. Urol. 36: 1341–1353, https://doi.org/10.1007/s00345-018-2286-7.Search in Google Scholar
Saleeb, R.M., Farag, M., Lichner, Z., Brimo, F., Bartlett, J., Bjarnason, G., Finelli, A., Rontondo, F., Downes, M.R., and Yousef, G.M. (2018). Modulating ATP binding cassette transporters in papillary renal cell carcinoma type 2 enhances its response to targeted molecular therapy. Mol. Oncol. 12: 1673–1688, https://doi.org/10.1002/1878-0261.12346.Search in Google Scholar
Seike, M., Kitamura, K., Miyanaga, A., Okano, T., Noro, R., and Gemma, A. (2014). Mir-134/487b/655 cluster regulates TGF-beta-induced epithelial-mesenchymal transition and drug resistance to gefitinib by targeting MAGI2 in lung adenocarcinoma cells. Cancer Res 74, https://doi.org/10.1158/1535-7163.MCT-13-0448.Search in Google Scholar
Tanji, N. and Yokoyama, M. (2011). Treatment of metastatic renal cell carcinoma and renal pelvic cancer. Clin. Exp. Nephrol. 15: 331–338, https://doi.org/10.1007/s10157-011-0438-9.Search in Google Scholar
Ulloa, L. and Tabibzadeh, S. (2001). Lefty inhibits receptor-regulated Smad phosphorylation induced by the activated transforming growth factor-β receptor. J. Biol. Chem. 276: 21397–21404, https://doi.org/10.1074/jbc.m010783200.Search in Google Scholar
van der Mijn, J.C., Mier, J.W., Broxterman, H.J., and Verheul, H.M. (2014). Predictive biomarkers in renal cell cancer: insights in drug resistance mechanisms. Drug Resist. Updates 17: 77–88, https://doi.org/10.1016/j.drup.2014.10.003.Search in Google Scholar
Wills, A.E. and Baker, J.C. (2015). E2a is necessary for Smad2/3-dependent transcription and the direct repression of Lefty during gastrulation. Dev. Cell 32: 345–357, https://doi.org/10.1016/j.devcel.2014.11.034.Search in Google Scholar
Xu, X.D., Zhang, L., He, X.G., Zhang, P., Sun, C.H., Xu, X.J., Lu, Y.J., and Li, F.F. (2018). TGF-beta plays a vital role in triple-negative breast cancer (TNBC) drug-resistance through regulating sternness, EMT and apoptosis. Biochem. Biophys. Res. Commun. 502: 160–165, https://doi.org/10.1016/j.bbrc.2018.05.139.Search in Google Scholar
Zabala, M., Lobo, N.A., Antony, J., Heitink, L.S., Gulati, G.S., Lam, J., Parashurama, N., Sanchez, K., Adorno, M., Sikandar, S.S., et al.. (2020). LEFTY1 is a dual-SMAD inhibitor that promotes mammary progenitor growth and tumorigenesis. Cell Stem Cell 27: 284–299 e288, https://doi.org/10.1016/j.stem.2020.06.017.Search in Google Scholar
Zhang, Z., Jiang, T., Li, Q., Wang, J., Yang, D., Li, X., Wang, Q., and Song, X. (2015). Nodal activates smad and extracellular signal-regulated kinases 1/2 pathways promoting renal cell carcinoma proliferation. Mol. Med. Rep. 12: 587–594, https://doi.org/10.3892/mmr.2015.3343.Search in Google Scholar
Zhou, N., Lu, F.D., Liu, C., Xu, K.W., Huang, J., Yu, D.X., and Bi, L.K. (2016). IL-8 induces the epithelial-mesenchymal transition of renal cell carcinoma cells through the activation of AKT signaling. Oncol. Lett. 12: 1915–1920, https://doi.org/10.3892/ol.2016.4900.Search in Google Scholar
© 2021 Walter de Gruyter GmbH, Berlin/Boston
