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Complementary Strand MicroRNAs Mediate Acquisition of Metastatic Potential in Colonic Adenocarcinoma

Journal of Gastrointestinal Surgery Aims and scope

Abstract

Background

Altered expression of specific microRNAs (miRNA) is known to occur during colorectal carcinogenesis. However, little is known about the genome-wide alterations in miRNA expression during the neoplastic progression of primary colorectal cancers.

Methods

Using a miRNA array platform, we evaluated the expression of 668 miRNA in primary colonic adenocarcinomas. Biological functions of selected miRNA were evaluated with in vitro invasion assays.

Results

RNA was extracted for miRNA analysis from 65 primary colon cancers. We identified a seven-miRNA expression signature that differentiated stage I and stage IV primary colon cancers. We then demonstrated this signature was able to discriminate between stage II and III primary colon cancers. Six differentially expressed miRNA were downregulated in association with the development of metastases, and all 7 miRNA were complementary strand miRNA. We transfected HCT-116, a highly invasive colon cancer cell line, with corresponding downregulated miRNA and demonstrated that overexpression of three miRNA (miR200c*, miR143*, and miR424*) significantly abrogated invasive potential.

Conclusion

We have identified a seven-miRNA signature that is associated with metastatic potential in the primary tumor. Forced overexpression of three downregulated miRNA resulted in attenuation of in vitro invasion, suggesting direct tumor suppressive function and further supporting the biological importance of complementary strand miRNA.

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References

  1. Jemal A, Siegel R, Xu J, Ward E. Cancer statistics, 2010. CA Cancer J Clin2010 Sep–Oct;60(5):277–300.

    Article  PubMed  Google Scholar 

  2. Glud M, Rossing M, Hother C, Holst L, Hastrup N, Nielsen FC, Gniadecki R, Drzewiecki KT. Downregulation of miR-125b in metastatic cutaneous malignant melanoma. Melanoma Res 2010 Dec;20(6):479–84.

    Article  PubMed  CAS  Google Scholar 

  3. Huynh C, Segura MF, Gaziel A, Menendez S, Darvishian F, Chiriboga L, Levine B, Meruelo D, Zavadil J, Marcusson EG, Hernando E. Efficient in vivo miRNA targeting of liver metastasis. Oncogene 2011 Mar 24;30(12):1481–8.

  4. Li Y, Zhang M, Chen H, Dong Z, Ganapathy V, Thangaraju M, Huang S. Ratio of miR-196s to HOXC8 messenger RNA correlates with breast cancer cell migration and metastasis. Cancer Res2010 Oct 15;70(20):7894–904.

    Article  PubMed  CAS  Google Scholar 

  5. Roth C, Rack B, Muller V, Janni W, Pantel K, Schwarzenbach H. Circulating microRNAs as blood-based markers for patients with primary and metastatic breast cancer. Breast Cancer Res2010 Nov 3;12(6):R90.

    Article  PubMed  CAS  Google Scholar 

  6. Su X, Chakravarti D, Cho MS, Liu L, Gi YJ, Lin YL, Leung ML, El-Naggar A, Creighton CJ, Suraokar MB, Wistuba I, Flores ER. TAp63 suppresses metastasis through coordinate regulation of Dicer and miRNAs. Nature2010 Oct 21;467(7318):986–90.

    Article  PubMed  CAS  Google Scholar 

  7. Faber C, Kirchner T, Hlubek F. The impact of microRNAs on colorectal cancer. Virchows Arch2009 Apr;454(4):359–67.

    Article  PubMed  CAS  Google Scholar 

  8. Czech B, Zhou R, Erlich Y, Brennecke J, Binari R, Villalta C, Gordon A, Perrimon N, Hannon GJ. Hierarchical rules for Argonaute loading in Drosophila. Mol Cell2009 Nov 13;36(3):445–56.

    Article  PubMed  CAS  Google Scholar 

  9. Ghildiyal M, Xu J, Seitz H, Weng Z, Zamore PD. Sorting of Drosophila small silencing RNAs partitions microRNA* strands into the RNA interference pathway. RNA2010 Jan;16(1):43–56.

    Article  PubMed  CAS  Google Scholar 

  10. Okamura K, Liu N, Lai EC. Distinct mechanisms for microRNA strand selection by Drosophila Argonautes. Mol Cell2009 Nov 13;36(3):431–44.

    Article  PubMed  CAS  Google Scholar 

  11. Xi Y, Formentini A, Chien M, Weir DB, Russo JJ, Ju J, Kornmann M. Prognostic values of microRNAs in colorectal cancer. Biomark Insights2006;2:113–21.

    PubMed  Google Scholar 

  12. Burk U, Schubert J, Wellner U, Schmalhofer O, Vincan E, Spaderna S, Brabletz T. A reciprocal repression between ZEB1 and members of the miR-200 family promotes EMT and invasion in cancer cells. EMBO Rep2008 Jun;9(6):582–9.

    Article  PubMed  CAS  Google Scholar 

  13. Tibshirani R, Hastie T, Narasimhan B, Chu G. Diagnosis of multiple cancer types by shrunken centroids of gene expression. Proc Natl Acad Sci U S A2002 May 14;99(10):6567–72.

    Article  PubMed  CAS  Google Scholar 

  14. Chen DT, Nasir A, Culhane A, Venkataramu C, Fulp W, Rubio R, Wang T, Agrawal D, McCarthy SM, Gruidl M, Bloom G, Anderson T, White J, Quackenbush J, Yeatman T. Proliferative genes dominate malignancy-risk gene signature in histologically-normal breast tissue. Breast Cancer Res Treat2010 Jan;119(2):335–46.

    Article  PubMed  Google Scholar 

  15. Baffa R, Fassan M, Volinia S, O'Hara B, Liu CG, Palazzo JP, Gardiman M, Rugge M, Gomella LG, Croce CM, Rosenberg A. MicroRNA expression profiling of human metastatic cancers identifies cancer gene targets. J Pathol2009 Oct;219(2):214–21.

    Article  PubMed  CAS  Google Scholar 

  16. Budhu A, Jia HL, Forgues M, Liu CG, Goldstein D, Lam A, Zanetti KA, Ye QH, Qin LX, Croce CM, Tang ZY, Wang XW. Identification of metastasis-related microRNAs in hepatocellular carcinoma. Hepatology2008 Mar;47(3):897–907.

    Article  PubMed  CAS  Google Scholar 

  17. Iorio MV, Croce CM. MicroRNAs in cancer: small molecules with a huge impact. J Clin Oncol2009 Dec 1;27(34):5848–56.

    Article  PubMed  CAS  Google Scholar 

  18. Lujambio A, Calin GA, Villanueva A, Ropero S, Sanchez-Cespedes M, Blanco D, Montuenga LM, Rossi S, Nicoloso MS, Faller WJ, Gallagher WM, Eccles SA, Croce CM, Esteller M. A microRNA DNA methylation signature for human cancer metastasis. Proc Natl Acad Sci U S A2008 Sep 9;105(36):13556–61.

    Article  PubMed  CAS  Google Scholar 

  19. Valastyan S, Reinhardt F, Benaich N, Calogrias D, Szasz AM, Wang ZC, Brock JE, Richardson AL, Weinberg RA. A pleiotropically acting microRNA, miR-31, inhibits breast cancer metastasis. Cell2009 Jun 12;137(6):1032–46.

    Article  PubMed  CAS  Google Scholar 

  20. Tsang WP, Kwok TT. The miR-18a* microRNA functions as a potential tumor suppressor by targeting on K-Ras. Carcinogenesis2009 Jun;30(6):953–9.

    Article  PubMed  CAS  Google Scholar 

  21. Kim S, Lee UJ, Kim MN, Lee EJ, Kim JY, Lee MY, Choung S, Kim YJ, Choi YC. MicroRNA miR-199a* regulates the MET proto-oncogene and the downstream extracellular signal-regulated kinase 2 (ERK2). J Biol Chem2008 Jun 27;283(26):18158–66.

    Article  PubMed  CAS  Google Scholar 

  22. Diederichs S, Haber DA. Dual role for argonautes in microRNA processing and posttranscriptional regulation of microRNA expression. Cell2007 Dec 14;131(6):1097–108.

    Article  PubMed  CAS  Google Scholar 

  23. Grishok A, Pasquinelli AE, Conte D, Li N, Parrish S, Ha I, Baillie DL, Fire A, Ruvkun G, Mello CC. Genes and mechanisms related to RNA interference regulate expression of the small temporal RNAs that control C. elegans developmental timing. Cell2001 Jul 13;106(1):23–34.

    Article  PubMed  CAS  Google Scholar 

  24. Khan AA, Betel D, Miller ML, Sander C, Leslie CS, Marks DS. Transfection of small RNAs globally perturbs gene regulation by endogenous microRNAs. Nat Biotechnol2009 Jun;27(6):549–55.

    PubMed  CAS  Google Scholar 

  25. O'Carroll D, Mecklenbrauker I, Das PP, Santana A, Koenig U, Enright AJ, Miska EA, Tarakhovsky A. A Slicer-independent role for Argonaute 2 in hematopoiesis and the microRNA pathway. Genes Dev2007 Aug 15;21(16):1999–2004.

    Article  PubMed  Google Scholar 

  26. Vaucheret H, Vazquez F, Crete P, Bartel DP. The action of ARGONAUTE1 in the miRNA pathway and its regulation by the miRNA pathway are crucial for plant development. Genes Dev2004 May 15;18(10):1187–97.

    Article  PubMed  CAS  Google Scholar 

  27. Ceppi P, Mudduluru G, Kumarswamy R, Rapa I, Scagliotti GV, Papotti M, Allgayer H. Loss of miR-200c expression induces an aggressive, invasive, and chemoresistant phenotype in non-small cell lung cancer. Mol Cancer Res2010 Sep;8(9):1207–16.

    Article  PubMed  CAS  Google Scholar 

  28. Elson-Schwab I, Lorentzen A, Marshall CJ. MicroRNA-200 family members differentially regulate morphological plasticity and mode of melanoma cell invasion. PLoS One2010;5(10).

  29. Hamano R, Miyata H, Yamasaki M, Kurokawa Y, Hara J, Ho Moon J, Nakajima K, Takiguchi S, Fujiwara Y, Mori M, Doki Y. Overexpression of miR-200c induces chemoresistance in esophageal cancers mediated through activation of the akt signaling pathway. Clin Cancer Res2011 Apr 19.

  30. Neves R, Scheel C, Weinhold S, Honisch E, Iwaniuk KM, Trompeter HI, Niederacher D, Wernet P, Santourlidis S, Uhrberg M. Role of DNA methylation in miR-200c/141 cluster silencing in invasive breast cancer cells. BMC Res Notes2010;3:219.

    Article  PubMed  Google Scholar 

  31. Smith CM, Watson DI, Leong MP, Mayne GC, Michael MZ, Wijnhoven BP, Hussey DJ. miR-200 Family expression is downregulated upon neoplastic progression of Barrett's esophagus. World J Gastroenterol2011 Feb 28;17(8):1036–44.

    PubMed  Google Scholar 

  32. Yu J, Ohuchida K, Mizumoto K, Sato N, Kayashima T, Fujita H, Nakata K, Tanaka M. MicroRNA, hsa-miR-200c, is an independent prognostic factor in pancreatic cancer and its upregulation inhibits pancreatic cancer invasion but increases cell proliferation. Mol Cancer2010;9:169.

    Article  PubMed  Google Scholar 

  33. Loboda A, Nebozhyn MV, Watters JW, Buser CA, Shaw PM, Huang PS, Van't Veer L, Tollenaar RA, Jackson DB, Agrawal D, Dai H, Yeatman TJ. EMT is the dominant program in human colon cancer. BMC Med Genomics2011;4:9.

    PubMed  Google Scholar 

  34. Liu W, Zabirnyk O, Wang H, Shiao YH, Nickerson ML, Khalil S, Anderson LM, Perantoni AO, Phang JM. miR-23b targets proline oxidase, a novel tumor suppressor protein in renal cancer. Oncogene2010 Sep 2;29(35):4914–24.

    Article  PubMed  CAS  Google Scholar 

  35. Jazdzewski K, Liyanarachchi S, Swierniak M, Pachucki J, Ringel MD, Jarzab B, de la Chapelle A. Polymorphic mature microRNAs from passenger strand of pre-miR-146a contribute to thyroid cancer. Proc Natl Acad Sci U S A2009 Feb 3;106(5):1502–5.

    Article  PubMed  CAS  Google Scholar 

  36. Schulte JH, Marschall T, Martin M, Rosenstiel P, Mestdagh P, Schlierf S, Thor T, Vandesompele J, Eggert A, Schreiber S, Rahmann S, Schramm A. Deep sequencing reveals differential expression of microRNAs in favorable versus unfavorable neuroblastoma. Nucleic Acids Res2010 Sep 1;38(17):5919–28.

    Article  PubMed  CAS  Google Scholar 

  37. Yang JS, Phillips MD, Betel D, Mu P, Ventura A, Siepel AC, Chen KC, Lai EC. Widespread regulatory activity of vertebrate microRNA* species. RNA2011 Feb;17(2):312–26.

    Article  PubMed  CAS  Google Scholar 

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Funding

National Cancer Institute Grant (CA112215).

Florida Department of Health Bankhead-Coley Cancer Program Grant (08BR-02).

Author information

Authors and Affiliations

  1. Department of Gastrointestinal Oncology, H. Lee Moffitt Cancer Center and Research Institute, 12902 Magnolia Drive, Tampa, FL, 33612, USA

    Jonathan M. Hernandez, David Shibata, Susan M. McCarthy, Leigh Ann Humphries, Whalen Clark, Abul Elahi, Mike Gruidl & Timothy Yeatman

  2. Department of Biomedical Informatics, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA

    Dung-Tsa Chen

  3. Department of Anatomic Pathology, H. Lee Moffitt Cancer Center and Research Institute, Tampa, FL, USA

    Domenico Coppola

  4. Department of Surgery, University of South Florida, Tampa, FL, USA

    Jonathan M. Hernandez & Whalen Clark

Authors
  1. Dung-Tsa Chen
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Correspondence to Jonathan M. Hernandez.

Additional information

Discussant

Yun Shin Chun (Philadelphia, PA): Dr. Hernandez, congratulations on this important contribution to our understanding of molecular changes in colon cancer. Since one microRNA (miRNA) regulates expression of hundreds of genes, much information can be gathered from analysis of over 600 miRNAs as you have shown. The seven miRNA signature significantly discriminates between stage I and IV colon cancer patients and between stage II and III, although there is no difference between stage III and IV patients. Six of the seven miRNAs are downregulated, consistent with published literature showing a general downregulation of miRNAs in cancer cells compared to non-tumorous tissue, indicating a tumor suppressor effect of many miRNAs. Notably, all of the seven miRNAs in this signature are complementary strand miRNAs, which were previously thought to be biologically inactive. There was no correlation between levels of complementary strand and mature miRNAs, suggesting a biologic function of complementary strand miRNAs distinct from their corresponding mature miRNAs.

I had three questions:

1. What are the differential levels of miRNAs in primary colon cancers compared with nodal and systemic metastases, and what are the changes in expression levels with preoperative chemotherapy?

2. Second, miR-21 is perhaps the most studied dysregulated miRNA in colon cancer. What were the expression levels of miR-21 in your patients?

3. Third, can you speculate on practical diagnostic or therapeutic applications of your findings?

Closing Discussant

Dr. Jonathan Hernandez: Regarding your first question, we have not evaluated the effect of neoadjuvant chemotherapy on the expression of miRNA. We have also not specifically evaluated miRNA expression in either lymph node or systemic metastases, although this would be of significant interest. If one accepts that lymph node metastases represent in-transit cells similar to circulating tumor cells, we would expect low or negligible expression of miRNA that inhibit genes associated with the acquisition of metastatic potential. According to our data, miR200c*, miR143*, and miR424* may be examples of such miRNA. If conceptualized in the context of epithelial-to-mesenchymal transition (EMT), we would predict that systemic macrometastases display miRNA expression profiles similar to primary tumors given that mobile mesenchymal cells undergo an epithelial transition in order to recapitulate a tumor mass.

To answer your second question, miR-21 has been shown to target PTEN, and thereby deregulate the PI3K pathway, increasing AKT signaling. We found no differential expression of miR-21 across the spectrum of AJCC stage I-IV primary colon cancers. We definitely observed expression of miR-21, although I’m not sure the absolute value is meaningful in terms of external comparison.

Finally, in response to your third question, our signature may have prognostic value in discriminating those patients with apparent early-stage tumors that eventually develop overt metastatic disease. We may be able to determine a metastatic probability for these patients based upon the expression of metastases-associated miRNA in the primary tumor. Firstly, such a signature may allow the identification of patients with early stage disease who may benefit from the use of adjuvant chemotherapy. Secondly, we believe that the therapeutic application as it relates to the individual miRNAs will be predicated upon elucidation of the functional biology of each miRNA in our signature. In this latter case, however, anti-miR therapy is currently in its infancy and may not have immediate direct translational applicability.

Dung-Tsa Chen and Jonathan M. Hernandez contributed equally to the work and share first authorship.

DC: Study concept and design, analysis of data, drafting of the manuscript, JMH: Study concept and design, interpretation of data, drafting of the manuscript, DS: Interpretation of data, drafting of the manuscript, SMM: Acquisition of data, LAH: Acquisition of data ,WC: Acquisition of data, AE: Acquisition of data, MG: Study concept and design, DC: Acquisition of data, TY: Study concept and design, critical revision of the manuscript, obtained funding

This work was conducted at Moffitt Cancer Center. We have no conflicts of interest to disclose.

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Chen, DT., Hernandez, J.M., Shibata, D. et al. Complementary Strand MicroRNAs Mediate Acquisition of Metastatic Potential in Colonic Adenocarcinoma. J Gastrointest Surg 16, 905–913 (2012). https://doi.org/10.1007/s11605-011-1815-0

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  • DOI: https://doi.org/10.1007/s11605-011-1815-0

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