Annals of Surgical Oncology

, Volume 21, Issue 3, pp 1046–1054 | Cite as

MicroRNAs with Prognostic Potential for Metastasis in Clear Cell Renal Cell Carcinoma: A Comparison of Primary Tumors and Distant Metastases

  • Joana Heinzelmann
  • André Unrein
  • Ulrike Wickmann
  • Sophie Baumgart
  • Marcus Stapf
  • Attila Szendroi
  • Marc-Oliver Grimm
  • Mieczyslaw R. Gajda
  • Heiko Wunderlich
  • Kerstin Junker
Urologic Oncology



MicroRNAs (miRNAs) are regulators of gene expression in tumor development and progression. However, their influence on metastasis of clear cell renal cell carcinoma (ccRCC) is less understood. To determine the role of miRNAs in metastatic progression, miRNA expression in primary ccRCC was compared to distant metastases.


Total RNA of 53 primary ccRCCs, 35 distant metastases from lung, bone, brain, and abdomen, as well as 17 normal kidney tissues was isolated from fresh frozen tissue and formalin-fixed paraffin-embedded (FFPE) samples. The miRNA microarrays were performed based on fresh frozen tissue. Results were validated by quantitative reverse transcription-polymerase chain reaction (qRT-PCR) on fresh frozen tissue and FFPE samples. Real-time cell analyses and transwell invasion assays were carried out after transient transfection of microRNA-30c (miR-30c) in cell line 786-O.


There were 14 miRNAs differently expressed in metastatic primary ccRCC and distant metastases compared to non-metastatic primary tumors. A strong correlation of miRNAs to progression-free- and cancer-specific 5-year-survival was determined. Specific miRNAs were differently expressed in distant metastases compared to primary ccRCC. A miRNA signature distinguished lung metastases from other metastatic sites. Overexpression of miR-30c increased adherence and decreased migration and invasion in the ccRCC cell line.


MiRNAs are deregulated in metastatic primary ccRCC and could be promising prognostic markers for an early prediction of metastasis. Alterations in miRNA expression characterize distant metastases of different metastatic sites. Furthermore, our study suggests a functional role of miR-30c in metastasis. The miRNAs could be a helpful tool for individual follow-up prediction and personalized therapy selection.


Renal Cell Carcinoma Distant Metastasis miRNA Expression Metastatic Site Clear Cell Renal Cell Carcinoma 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.



This study was supported by a grant from Dr. Robert Pfleger-Stiftung. We also thank Dr. Saiful Miah, Dr. Matthias Saar, and Dr. Sebastian Hölters for their critical reading of this manuscript.


The authors declare that they have no conflict of interest.


  1. 1.
    Bartel DP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116(2):281–97.PubMedCrossRefGoogle Scholar
  2. 2.
    Croce CM, Calin GA. miRNAs, cancer, and stem cell division. Cell 2005;122(1):6–7. doi: 10.1016/j.cell.2005.06.036.PubMedCrossRefGoogle Scholar
  3. 3.
    Huang ZM, Yang J, Shen XY, et al. MicroRNA expression profile in non-cancerous colonic tissue associated with lymph node metastasis of colon cancer. J Dig Dis. 2009;10(3):188–94. doi: 10.1111/j.1751-2980.2009.00384.x.PubMedCrossRefGoogle Scholar
  4. 4.
    Junker K, Ficarra V, Kwon ED, Leibovich BC, Thompson RH, Oosterwijk E. Potential role of genetic markers in the management of kidney cancer. Eur Urol. 2013;63(2):333–40. doi: 10.1016/j.eururo.2012.09.040.PubMedCrossRefGoogle Scholar
  5. 5.
    Jung M, Mollenkopf HJ, Grimm C, et al. MicroRNA profiling of clear cell renal cell cancer identifies a robust signature to define renal malignancy. J Cell Mol Med. 2009;13(9B):3918–28. doi: 10.1111/j.1582-4934.2009.00705.x.PubMedCrossRefGoogle Scholar
  6. 6.
    Juan D, Alexe G, Antes T, et al. Identification of a microRNA panel for clear-cell kidney cancer. Urology. 2010;75(4):835–41. doi: 10.1016/j.urology.2009.10.033.PubMedCrossRefGoogle Scholar
  7. 7.
    Nakada C, Matsuura K, Tsukamoto Y, et al. Genome-wide microRNA expression profiling in renal cell carcinoma: significant down-regulation of miR-141 and miR-200c. J Pathol. 2008;216(4):417–27. doi: 10.1002/path.2437.CrossRefGoogle Scholar
  8. 8.
    Petillo D, Kort EJ, Anema J, Furge KA, Yang XJ, Teh BT. MicroRNA profiling of human kidney cancer subtypes. Int J Oncol. 2009;35(1):109–14.PubMedCrossRefGoogle Scholar
  9. 9.
    Youssef YM, White NM, Grigull J, et al. Accurate molecular classification of kidney cancer subtypes using microRNA signature. Eur Urol. 2011;59(5)721–30. doi: 10.1016/j.eururo.2011.01.004.PubMedCrossRefGoogle Scholar
  10. 10.
    Gottardo F, Liu CG, Ferracin M, et al. Micro-RNA profiling in kidney and bladder cancers. Urol Oncol. 2007;25:387–92. doi: 10.1016/j.urolonc.2007.01.019.PubMedCrossRefGoogle Scholar
  11. 11.
    Chow TF, Youssef YM, Lianidou E, et al. Differential expression profiling of microRNAs and their potential involvement in renal cell carcinoma pathogenesis. Clin Biochem. 2010;43(1–2):150–8. doi: 10.1016/j.clinbiochem.2009.07.020.PubMedCrossRefGoogle Scholar
  12. 12.
    Storkel S, van den Berg E. Morphological classification of renal cancer. World J Urol. 1995;13(3):153–8.PubMedCrossRefGoogle Scholar
  13. 13.
    Heinzelmann J, Henning B, Sanjmyatav J, et al. Specific miRNA signatures are associated with metastasis and poor prognosis in clear cell renal cell carcinoma. World J Urol. 2011;29(3):367–73. doi: 10.1007/s00345-010-0633-4.PubMedCrossRefGoogle Scholar
  14. 14.
    Saito K, Oku T, Ata N, Miyashiro H, Hattori M, Saiki I. A modified and convenient method for assessing tumor cell invasion and migration and its application to screening for inhibitors. Biol Pharm Bull. 1997;20(4):345–8.PubMedCrossRefGoogle Scholar
  15. 15.
    Calin GA, Croce CM. MicroRNA signatures in human cancers. Nat Rev Cancer. 2006;6(11):857–66. doi: 10.1038/nrc1997.PubMedCrossRefGoogle Scholar
  16. 16.
    Esquela-Kerscher A, Slack FJ. Oncomirs—microRNAs with a role in cancer. Nat Rev Cancer. 2006;6(4):259–69. doi: 10.1038/nrc1840.PubMedCrossRefGoogle Scholar
  17. 17.
    Catto JW, Alcaraz A, Bjartell AS, et al. MicroRNA in prostate, bladder, and kidney cancer: a systematic review. Eur Urol. 2011;59(5):671–81. doi: 10.1016/j.eururo.2011.01.044.PubMedCrossRefGoogle Scholar
  18. 18.
    Williams RD, Elliott AY, Stein N, Fraley EE. In vitro cultivation of human renal cell cancer. II. Characterization of cell lines. In Vitro. 1978;14(9):779–6.PubMedCrossRefGoogle Scholar
  19. 19.
    Boominathan L. The guardians of the genome (p53, TA-p73, and TA-p63) are regulators of tumor suppressor miRNAs network. Cancer Metastasis Rev. 2010;29(4);613–39. doi: 10.1007/s10555-010-9257-9.PubMedCrossRefGoogle Scholar
  20. 20.
    Braun J, Hoang-Vu C, Dralle H, Huttelmaier S. Downregulation of microRNAs directs the EMT and invasive potential of anaplastic thyroid carcinomas. Oncogene. 2010;29(29):4237–44. doi: 10.1038/onc.2010.169.PubMedCrossRefGoogle Scholar
  21. 21.
    Khella H, White N, Faragalla H, et al. Exploring the role of miRNAs in renal cell carcinoma progression and metastasis through bioinformatic and experimental analyses. Tumor Biol. 2012;33(1):131–40. doi: 10.1007/s13277-011-0255-5.CrossRefGoogle Scholar
  22. 22.
    White NM, Khella HW, Grigull J, et al. miRNA profiling in metastatic renal cell carcinoma reveals a tumour-suppressor effect for miR-215. Br J Cancer. 2011;105(11):1741–9. doi: 10.1038/bjc.2011.401.PubMedCentralPubMedCrossRefGoogle Scholar
  23. 23.
    Feng R, Chen X, Yu Y, et al. miR-126 functions as a tumour suppressor in human gastric cancer. Cancer Lett. 2010;298(1):50–63. doi: 10.1016/j.canlet.2010.06.004.PubMedCrossRefGoogle Scholar
  24. 24.
    Donnem T, Lonvik K, Eklo K, et al. Independent and tissue-specific prognostic impact of miR-126 in nonsmall cell lung cancer: coexpression with vascular endothelial growth factor-A predicts poor survival. Cancer. 2011;117(14):3193–200. doi: 10.1002/cncr.25907.PubMedCrossRefGoogle Scholar
  25. 25.
    Tavazoie SF, Alarcon C, Oskarsson T, et al. Endogenous human microRNAs that suppress breast cancer metastasis. Nature. 2008;451(7175):147–52. doi: 10.1038/nature06487.PubMedCentralPubMedCrossRefGoogle Scholar
  26. 26.
    Wotschofsky Z, Liep J, Meyer HA, et al. Identification of metastamirs as metastasis-associated microRNAs in clear cell renal cell carcinomas. Int J Biol Sci. 2012;8(10):1363–74. doi: 10.7150/ijbs.5106.PubMedCentralPubMedCrossRefGoogle Scholar
  27. 27.
    Wu X, Weng L, Li X, et al. Identification of a 4-microRNA Signature for Clear Cell Renal Cell Carcinoma Metastasis and Prognosis. PLoS ONE. 2012;7(5):e35661. doi: 10.1371/journal.pone.0035661.PubMedCentralPubMedCrossRefGoogle Scholar
  28. 28.
    Ma L, Teruya-Feldstein J, Weinberg RA. Tumour invasion and metastasis initiated by microRNA-10b in breast cancer. Nature. 2007;449(7163):682–8. doi: 10.1038/nature06174.PubMedCrossRefGoogle Scholar
  29. 29.
    Liu Z, Zhu J, Cao H, Ren H, Fang X. miR-10b promotes cell invasion through RhoC-AKT signaling pathway by targeting HOXD10 in gastric cancer. Int J Oncol. 2012;40(5):1553–60. doi: 10.3892/ijo.2012.1342.PubMedGoogle Scholar
  30. 30.
    Lin J, Teo S, Lam DH, Jeyaseelan K, Wang S. MicroRNA-10b pleiotropically regulates invasion, angiogenicity and apoptosis of tumor cells resembling mesenchymal subtype of glioblastoma multiforme. Cell Death Dis. 2012;3:e398. doi: 10.1038/cddis.2012.134.PubMedCentralPubMedCrossRefGoogle Scholar

Copyright information

© Society of Surgical Oncology 2013

Authors and Affiliations

  • Joana Heinzelmann
    • 1
    • 2
  • André Unrein
    • 2
  • Ulrike Wickmann
    • 2
  • Sophie Baumgart
    • 1
    • 2
  • Marcus Stapf
    • 2
  • Attila Szendroi
    • 3
  • Marc-Oliver Grimm
    • 2
  • Mieczyslaw R. Gajda
    • 4
  • Heiko Wunderlich
    • 5
    • 2
  • Kerstin Junker
    • 1
    • 2
  1. 1.Clinic of Urology and Pediatric UrologySaarland University Medical CenterHomburg/SaarGermany
  2. 2.Department of UrologyJena University HospitalJenaGermany
  3. 3.Clinic of UrologySemmelweis University BudapestBudapestHungary
  4. 4.Institute of PathologyJena University HospitalJenaGermany
  5. 5.Clinic of Urology and Pediatric UrologyEisenach St. Georg HospitalEisenachGermany

Personalised recommendations