Skip to main content

Advertisement

SpringerLink
Log in

MicroRNA-30a inhibits cell migration and invasion by downregulating vimentin expression and is a potential prognostic marker in breast cancer

  • Preclinical Study
  • Published:
Breast Cancer Research and Treatment Aims and scope Submit manuscript

Abstract

Tumor recurrence and metastasis result in an unfavorable prognosis for cancer patients. Recent studies have suggested that specific microRNAs (miRNAs) may play important roles in the development of cancer cells. However, prognostic markers and the outcome prediction of the miRNA signature in breast cancer patients have not been comprehensively assessed. The aim of this study was to identify miRNA biomarkers relating to clinicopathological features and outcome of breast cancer. A miRNA microarray analysis was performed on breast tumors of different lymph node metastasis status and with different progression signatures, indicated by overexpression of cyclin D1 and β-catenin genes, to identify miRNAs showing a significant difference in expression. The functional interaction between the candidate miRNA, miR-30a, and the target gene, Vim, which codes for vimentin, a protein involved in epithelial–mesenchymal transition, was examined using the luciferase reporter assay, western blotting, and migration and invasion assays. The association between the decreased miR-30a levels and breast cancer progression was examined in a survival analysis. miR-30a negatively regulated vimentin expression by binding to the 3′-untranslated region of Vim. Overexpression of miR-30a suppressed the migration and invasiveness phenotypes of breast cancer cell lines. Moreover, reduced tumor expression of miR-30a in breast cancer patients was associated with an unfavorable outcome, including late tumor stage, lymph node metastasis, and worse progression (mortality and recurrence) (p < 0.05). In conclusion, these findings suggest a role for miR-30a in inhibiting breast tumor invasiveness and metastasis. The finding that miR-30a downmodulates vimentin expression might provide a therapeutic target for the treatment of breast cancer.

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

Access this article

Log in via an institution

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

Similar content being viewed by others

Abbreviations

IDC:

Invasive ductal carcinoma

EMT:

Epithelial–mesenchymal transition

miRNA:

microRNA

3′UTR:

3′-untranslated region

LNM:

Lymph node metastasis

LCM:

Laser capture microdissection

qRT-PCR:

Quantitative real-time reverse transcription polymerase chain reaction

DFS:

Disease-free survival

OS:

Overall survival

HR:

Hazard ratio

OR:

Odds ratio

95 % CI:

95 % confidence interval

References

  1. Orlando FA, Brown KD (2009) Unraveling breast cancer heterogeneity through transcriptomic and epigenomic analysis. Ann Surg Oncol 16(8):2270–2279

    Article  PubMed  Google Scholar 

  2. Polyak K (2007) Breast cancer: origins and evolution. J Clin Invest 117(11):3155–3163

    Article  PubMed  CAS  Google Scholar 

  3. Guarino M, Rubino B, Ballabio G (2007) The role of epithelial–mesenchymal transition in cancer pathology. Pathology 39(3):305–318

    Article  PubMed  CAS  Google Scholar 

  4. Micalizzi DS, Farabaugh SM, Ford HL (2010) Epithelial–mesenchymal transition in cancer: parallels between normal development and tumor progression. J Mammary Gland Biol Neoplasia 15(2):117–134

    Article  PubMed  Google Scholar 

  5. Trimboli AJ, Fukino K, de Bruin A, Wei G, Shen L, Tanner SM, Creasap N, Rosol TJ, Robinson ML, Eng C, Ostrowski MC, Leone G (2008) Direct evidence for epithelial–mesenchymal transitions in breast cancer. Cancer Res 68(3):937–945

    Article  PubMed  CAS  Google Scholar 

  6. Wells A, Yates C, Shepard CR (2008) E-cadherin as an indicator of mesenchymal to epithelial reverting transitions during the metastatic seeding of disseminated carcinomas. Clin Exp Metastasis 25(6):621–628

    Article  PubMed  CAS  Google Scholar 

  7. Marsit CJ, Posner MR, McClean MD, Kelsey KT (2008) Hypermethylation of E-cadherin is an independent predictor of improved survival in head and neck squamous cell carcinoma. Cancer 113(7):1566–1571

    Article  PubMed  Google Scholar 

  8. Prasad CP, Mirza S, Sharma G, Prashad R, DattaGupta S, Rath G, Ralhan R (2008) Epigenetic alterations of CDH1 and APC genes: relationship with activation of Wnt/beta-catenin pathway in invasive ductal carcinoma of breast. Life Sci 83(9–10):318–325

    Article  PubMed  CAS  Google Scholar 

  9. Yates DR, Rehman I, Abbod MF, Meuth M, Cross SS, Linkens DA, Hamdy FC, Catto JW (2007) Promoter hypermethylation identifies progression risk in bladder cancer. Clin Cancer Res 13(7):2046–2053

    Article  PubMed  CAS  Google Scholar 

  10. Graziano F, Humar B, Guilford P (2003) The role of the E-cadherin gene (CDH1) in diffuse gastric cancer susceptibility: from the laboratory to clinical practice. Ann Oncol 14(12):1705–1713

    Article  PubMed  CAS  Google Scholar 

  11. Braun J, Hoang-Vu C, Dralle H, Huttelmaier S (2010) Downregulation of microRNAs directs the EMT and invasive potential of anaplastic thyroid carcinomas. Oncogene 29(29):4237–4244

    Article  PubMed  CAS  Google Scholar 

  12. Vetter G, Saumet A, Moes M, Vallar L, Le Bechec A, Laurini C, Sabbah M, Arar K, Theillet C, Lecellier CH, Friederich E (2010) miR-661 expression in SNAI1-induced epithelial to mesenchymal transition contributes to breast cancer cell invasion by targeting Nectin-1 and StarD10 messengers. Oncogene 29(31):4436–4448

    Article  PubMed  CAS  Google Scholar 

  13. Cho WC (2007) OncomiRs: the discovery and progress of microRNAs in cancers. Mol Cancer 6:60

    Article  PubMed  Google Scholar 

  14. Negrini M, Nicoloso MS, Calin GA (2009) MicroRNAs and cancer—new paradigms in molecular oncology. Curr Opin Cell Biol 21(3):470–479

    Article  PubMed  CAS  Google Scholar 

  15. Ortholan C, Puissegur MP, Ilie M, Barbry P, Mari B, Hofman P (2009) MicroRNAs and lung cancer: new oncogenes and tumor suppressors, new prognostic factors and potential therapeutic targets. Curr Med Chem 16(9):1047–1061

    Article  PubMed  CAS  Google Scholar 

  16. Shenouda SK, Alahari SK (2009) MicroRNA function in cancer: oncogene or a tumor suppressor? Cancer Metastasis Rev 28(3–4):369–378

    Article  PubMed  CAS  Google Scholar 

  17. Iwatsuki M, Mimori K, Fukagawa T, Ishii H, Yokobori T, Sasako M, Baba H, Mori M (2010) The clinical significance of vimentin-expressing gastric cancer cells in bone marrow. Ann Surg Oncol 17(9):2526–2533

    Article  PubMed  Google Scholar 

  18. Mendez MG, Kojima S, Goldman RD (2010) Vimentin induces changes in cell shape, motility, and adhesion during the epithelial to mesenchymal transition. FASEB J 24(6):1838–1851

    Article  PubMed  CAS  Google Scholar 

  19. Usami Y, Satake S, Nakayama F, Matsumoto M, Ohnuma K, Komori T, Semba S, Ito A, Yokozaki H (2008) Snail-associated epithelial–mesenchymal transition promotes oesophageal squamous cell carcinoma motility and progression. J Pathol 215(3):330–339

    Article  PubMed  CAS  Google Scholar 

  20. Yang PS, Yang TL, Liu CL, Wu CW, Shen CY (1997) A case-control study of breast cancer in Taiwan—a low-incidence area. Br J Cancer 75(5):752–756

    Article  PubMed  CAS  Google Scholar 

  21. Lo YL, Yu JC, Huang CS, Tseng SL, Chang TM, Chang KJ, Wu CW, Shen CY (1998) Allelic loss of the BRCA1 and BRCA2 genes and other regions on 17q and 13q in breast cancer among women from Taiwan (area of low incidence but early onset). Int J Cancer 79(6):580–587

    Article  PubMed  CAS  Google Scholar 

  22. Cheng TC, Chen ST, Huang CS, Fu YP, Yu JC, Cheng CW, Wu PE, Shen CY (2005) Breast cancer risk associated with genotype polymorphism of the catechol estrogen-metabolizing genes: a multigenic study on cancer susceptibility. Int J Cancer 113(3):345–353

    Article  PubMed  CAS  Google Scholar 

  23. Ming-Shiean H, Yu JC, Wang HW, Chen ST, Hsiung CN, Ding SL, Wu PE, Shen CY, Cheng CW (2010) Synergistic effects of polymorphisms in DNA repair genes and endogenous estrogen exposure on female breast cancer risk. Ann Surg Oncol 17(3):760–771

    Article  PubMed  Google Scholar 

  24. Ding SL, Sheu LF, Yu JC, Yang TL, Chen BF, Leu FJ, Shen CY (2004) Abnormality of the DNA double-strand-break checkpoint/repair genes, ATM, BRCA1 and TP53, in breast cancer is related to tumour grade. Br J Cancer 90(10):1995–2001

    Article  PubMed  CAS  Google Scholar 

  25. Hsu HM, Wang HC, Chen ST, Hsu GC, Shen CY, Yu JC (2007) Breast cancer risk is associated with the genes encoding the DNA double-strand break repair Mre11/Rad50/Nbs1 complex. Cancer Epidemiol Biomarkers Prev 16(10):2024–2032

    Article  PubMed  CAS  Google Scholar 

  26. Shen CY, Yu JC, Lo YL, Kuo CH, Yue CT, Jou YS, Huang CS, Lung JC, Wu CW (2000) Genome-wide search for loss of heterozygosity using laser capture microdissected tissue of breast carcinoma: an implication for mutator phenotype and breast cancer pathogenesis. Cancer Res 60(14):3884–3892

    PubMed  CAS  Google Scholar 

  27. Lo YL, Shen CY (2002) Laser capture microdissection in carcinoma analysis. Methods Enzymol 356:137–144

    Article  PubMed  Google Scholar 

  28. Petroff BK, Phillips TA, Kimler BF, Fabian CJ (2006) Detection of biomarker gene expression by real-time polymerase chain reaction using amplified ribonucleic acids from formalin-fixed random periareolar fine needle aspirates of human breast tissue. Anal Quant Cytol Histol 28(5):297–302

    PubMed  Google Scholar 

  29. Cheng CW, Yu JC, Wang HW, Huang CS, Shieh JC, Fu YP, Chang CW, Wu PE, Shen CY (2010) The clinical implications of MMP-11 and CK-20 expression in human breast cancer. Clin Chim Acta 411(3–4):234–241

    Article  PubMed  CAS  Google Scholar 

  30. Pfaffl MW (2001) A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res 29(9):e45

    Article  PubMed  CAS  Google Scholar 

  31. McInroy L, Maatta A (2007) Down-regulation of vimentin expression inhibits carcinoma cell migration and adhesion. Biochem Biophys Res Commun 360(1):109–114

    Article  PubMed  CAS  Google Scholar 

  32. Satelli A, Li S (2011) Vimentin in cancer and its potential as a molecular target for cancer therapy. Cell Mol Life Sci 68(18):3033–3046

    Article  PubMed  CAS  Google Scholar 

  33. Vuoriluoto K, Haugen H, Kiviluoto S, Mpindi JP, Nevo J, Gjerdrum C, Tiron C, Lorens JB, Ivaska J (2011) Vimentin regulates EMT induction by Slug and oncogenic H-Ras and migration by governing Axl expression in breast cancer. Oncogene 30(12):1436–1448

    Article  PubMed  CAS  Google Scholar 

  34. Franke WW, Grund C, Kuhn C, Jackson BW, Illmensee K (1982) Formation of cytoskeletal elements during mouse embryogenesis. III. Primary mesenchymal cells and the first appearance of vimentin filaments. Differentiation 23(1):43–59

    Article  PubMed  CAS  Google Scholar 

  35. Dutsch-Wicherek M, Lazar A, Tomaszewska R (2010) The potential role of MT and vimentin immunoreactivity in the remodeling of the microenvironment of parotid adenocarcinoma. Cancer Microenviron 4(1):105–113

    Article  PubMed  Google Scholar 

  36. Sarrio D, Palacios J, Hergueta-Redondo M, Gomez-Lopez G, Cano A, Moreno-Bueno G (2009) Functional characterization of E- and P-cadherin in invasive breast cancer cells. BMC Cancer 9:74

    Article  PubMed  Google Scholar 

  37. Mellios N, Huang HS, Grigorenko A, Rogaev E, Akbarian S (2008) A set of differentially expressed miRNAs, including miR-30a-5p, act as post-transcriptional inhibitors of BDNF in prefrontal cortex. Hum Mol Genet 17(19):3030–3042

    Article  PubMed  CAS  Google Scholar 

  38. Hand NJ, Master ZR, Eauclaire SF, Weinblatt DE, Matthews RP, Friedman JR (2009) The microRNA-30 family is required for vertebrate hepatobiliary development. Gastroenterology 136(3):1081–1090

    Article  PubMed  CAS  Google Scholar 

  39. Agrawal R, Tran U, Wessely O (2009) The miR-30 miRNA family regulates Xenopus pronephros development and targets the transcription factor Xlim1/Lhx1. Development 136(23):3927–3936

    Article  PubMed  CAS  Google Scholar 

  40. Kumarswamy R, Mudduluru G, Ceppi P, Muppala S, Kozlowski M, Niklinski J, Papotti M, Allgayer H (2012) MicroRNA-30a inhibits epithelial-to-mesenchymal transition by targeting Snai1 and is downregulated in non-small cell lung cancer. Int J Cancer 130(9):2044–2053

    Article  PubMed  CAS  Google Scholar 

  41. Izzotti A, Calin GA, Arrigo P, Steele VE, Croce CM, De Flora S (2009) Downregulation of microRNA expression in the lungs of rats exposed to cigarette smoke. FASEB J 23(3):806–812

    Article  PubMed  CAS  Google Scholar 

  42. Volinia S, Galasso M, Costinean S, Tagliavini L, Gamberoni G, Drusco A, Marchesini J, Mascellani N, Sana ME, Abu Jarour R, Desponts C, Teitell M, Baffa R, Aqeilan R, Iorio MV, Taccioli C, Garzon R, Di Leva G, Fabbri M, Catozzi M, Previati M, Ambs S, Palumbo T, Garofalo M, Veronese A, Bottoni A, Gasparini P, Harris CC, Visone R, Pekarsky Y, de la Chapelle A, Bloomston M, Dillhoff M, Rassenti LZ, Kipps TJ, Huebner K, Pichiorri F, Lenze D, Cairo S, Buendia MA, Pineau P, Dejean A, Zanesi N, Rossi S, Calin GA, Liu CG, Palatini J, Negrini M, Vecchione A, Rosenberg A, Croce CM (2010) Reprogramming of miRNA networks in cancer and leukemia. Genome Res 20(5):589–599

    Article  PubMed  CAS  Google Scholar 

  43. Chappell SA, Walsh T, Walker RA, Shaw JA (1997) Loss of heterozygosity at chromosome 6q in preinvasive and early invasive breast carcinomas. Br J Cancer 75(9):1324–1329

    Article  PubMed  CAS  Google Scholar 

  44. Noviello C, Courjal F, Theillet C (1996) Loss of heterozygosity on the long arm of chromosome 6 in breast cancer: possibly four regions of deletion. Clin Cancer Res 2(9):1601–1606

    PubMed  CAS  Google Scholar 

  45. Heinzelmann J, Henning B, Sanjmyatav J, Posorski N, Steiner T, Wunderlich H, Gajda MR, Junker K (2011) Specific miRNA signatures are associated with metastasis and poor prognosis in clear cell renal cell carcinoma. World J Urol 29(3):367–373

    Article  PubMed  CAS  Google Scholar 

  46. Li X, Zhang Y, Ding J, Wu K, Fan D (2010) Survival prediction of gastric cancer by a seven-microRNA signature. Gut 59(5):579–585

    Article  PubMed  CAS  Google Scholar 

  47. Tan X, Qin W, Zhang L, Hang J, Li B, Zhang C, Wan J, Zhou F, Shao K, Sun Y, Wu J, Zhang X, Qiu B, Li N, Shi S, Feng X, Zhao S, Wang Z, Zhao X, Chen Z, Mitchelson K, Cheng J, Guo Y, He J (2011) A 5-microRNA signature for lung squamous cell carcinoma diagnosis and hsa-miR-31 for prognosis. Clin Cancer Res 17(21):6802–6811

    Article  PubMed  CAS  Google Scholar 

  48. Brennecke J, Stark A, Russell RB, Cohen SM (2005) Principles of microRNA-target recognition. PLoS Biol 3(3):e85

    Article  PubMed  Google Scholar 

  49. Peter ME (2010) Targeting of mRNAs by multiple miRNAs: the next step. Oncogene 29(15):2161–2164

    Article  PubMed  CAS  Google Scholar 

  50. Ozcan S (2009) MiR-30 family and EMT in human fetal pancreatic islets. Islets 1(3):283–285

    Article  PubMed  Google Scholar 

  51. Kong W, Yang H, He L, Zhao JJ, Coppola D, Dalton WS, Cheng JQ (2008) MicroRNA-155 is regulated by the transforming growth factor beta/Smad pathway and contributes to epithelial cell plasticity by targeting RhoA. Mol Cell Biol 28(22):6773–6784

    Article  PubMed  CAS  Google Scholar 

  52. Wendt MK, Allington TM, Schiemann WP (2009) Mechanisms of the epithelial–mesenchymal transition by TGF-beta. Future Oncol 5(8):1145–1168

    Article  PubMed  CAS  Google Scholar 

  53. Rodriguez-Gonzalez FG, Sieuwerts AM, Smid M, Look MP, Meijer-van Gelder ME, de Weerd V, Sleijfer S, Martens JW, Foekens JA (2011) MicroRNA-30c expression level is an independent predictor of clinical benefit of endocrine therapy in advanced estrogen receptor positive breast cancer. Breast Cancer Res Treat 127(1):43–51

    Article  PubMed  CAS  Google Scholar 

Download references

Acknowledgments

We sincerely appreciate Ms. Show-Lin Yang for her assistance in organizing our study specimens. This study was supported by research grant NSC 98-2314-B-040-009-MY3 from the National Science Council, Taipei, Taiwan.

Conflict of interest

The authors declare that they have no conflict of interest.

Author information

Authors and Affiliations

  1. Institute of Biochemistry and Biotechnology, Chung Shan Medical University, Taichung, 40201, Taiwan

    Chun-Wen Cheng, Chia-Wei Chang & Cheng-You Chen

  2. Clinical Laboratory, Chung Shan Medical University Hospital, Taichung, Taiwan

    Chun-Wen Cheng

  3. Institute of Biomedical Sciences, Academia Sinica, Taipei, 11529, Taiwan

    Chun-Wen Cheng, Hsiao-Wei Wang, Hou-Wei Chu & Chen-Yang Shen

  4. Department of Surgery, Tri-Service General Hospital, National Defense Medical Center, Taipei, Taiwan

    Jyh-Cherng Yu

  5. Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan

    Jui-I Chao & Huei-Fang Liu

  6. Department of Nursing, Kang-Ning Junior College of Medical Care and Management, Taipei, Taiwan

    Shian-ling Ding

  7. Graduate Institute of Environmental Science, China Medical University, Taichung, Taiwan

    Chen-Yang Shen

Authors
  1. Chun-Wen Cheng
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hsiao-Wei Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chia-Wei Chang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Hou-Wei Chu
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Cheng-You Chen
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jyh-Cherng Yu
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jui-I Chao
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Huei-Fang Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Shian-ling Ding
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Chen-Yang Shen
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Chun-Wen Cheng or Chen-Yang Shen.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Cheng, CW., Wang, HW., Chang, CW. et al. MicroRNA-30a inhibits cell migration and invasion by downregulating vimentin expression and is a potential prognostic marker in breast cancer. Breast Cancer Res Treat 134, 1081–1093 (2012). https://doi.org/10.1007/s10549-012-2034-4

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10549-012-2034-4

Keywords

Search

Navigation