Skip to main content
SpringerLink
Log in

MicroRNA dysregulation in adenoid cystic carcinoma of the salivary gland in relation to prognosis and gene fusion status: a cohort study

  • Original Article
  • Published:
Virchows Archiv Aims and scope Submit manuscript

Abstract

Adenoid cystic carcinoma (ACC) is among the most frequent malignancies of the salivary gland, and is notorious for its prolonged clinical course characterized by frequent recurrences often years after initial treatment. No molecular marker has been shown to have independent prognostic value in ACC, including characteristic gene fusions involving MYB, MYBL1, and NFIB. MicroRNA has been shown to be associated with clinical outcome in numerous malignancies, including one study of ACC, warranting further validation of this class of markers in this disease. Here, we investigate the prognostic value of microRNA in two ACC cohorts: a training cohort (n = 64) and a validation cohort (n = 120) with microarray and qPCR. In the training cohort, multivariate analysis of microarray data found high expression of hsa-miR-6835-3p to be associated with reduced recurrence-free survival (RFS) (p = 0.016). Measuring the highest ranking microRNAs identified in survival analysis in the same cohort, qPCR identified high expression of hsa-miR-4676 to be associated with reduced overall survival (OS) and high expression of hsa-mir-1180 to be associated with improved RFS. This was not confirmed in the validation cohort, in which qPCR identified high expression of hsa-mir-21, hsa-mir-181a-2, and hsa-mir-152 to be associated with reduced OS and high expression of hsa-miR-374c to be associated with improved RFS. Interestingly, two distinct subsets of ACC separated in microRNA expression irrespective of gene fusion status, but without significant difference in outcome. Collectively, qPCR identified several microRNAs associated with OS and RFS, and different subsets of ACC separated according to microRNA expression, suggestive of ACC being a heterogeneous group of malignancies in its microRNA profile.

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

References

  1. Bjørndal K, Krogdahl A, Therkildsen MH, Charabi B, Kristensen CA, Andersen E, Schytte S, Primdahl H, Johansen J, Pedersen HB, Andersen LJ, Godballe C (2015) Salivary adenoid cystic carcinoma in Denmark 1990–2005: Outcome and independent prognostic factors including the benefit of radiotherapy. Results of the Danish head and neck Cancer group (DAHANCA). Oral Oncol 51:1138–1142. https://doi.org/10.1016/j.oraloncology.2015.10.002

    Article  PubMed  Google Scholar 

  2. Kokemueller H, Eckardt A, Brachvogel P, Hausamen JE (2004) Adenoid cystic carcinoma of the head and neck—a 20 years experience. Int J Oral Maxillofac Surg 33:25–31. https://doi.org/10.1054/ijom.2003.0448

    Article  PubMed  CAS  Google Scholar 

  3. Oplatek A, Ozer E, Agrawal A, Bapna S, Schuller DE (2010) Patterns of recurrence and survival of head and neck adenoid cystic carcinoma after definitive resection. Laryngoscope 120:65–70. https://doi.org/10.1002/lary.20684

    Article  PubMed  Google Scholar 

  4. Chen AM, Bucci MK, Weinberg V, Garcia J, Quivey JM, Schechter NR, Phillips TL, Fu KK, Eisele DW (2006) Adenoid cystic carcinoma of the head and neck treated by surgery with or without postoperative radiation therapy: Prognostic features of recurrence. Int J Radiat Oncol Biol Phys 66:152–159. https://doi.org/10.1016/j.ijrobp.2006.04.014

    Article  PubMed  Google Scholar 

  5. Balamucki CJ, Amdur RJ, Werning JW, Vaysberg M, Morris CG, Kirwan JM, Mendenhall WM (2012) Adenoid cystic carcinoma of the head and neck. Am J Otolaryngol 33:510–518. https://doi.org/10.1016/j.amjoto.2011.11.006

    Article  PubMed  Google Scholar 

  6. Adelstein DJ, Koyfman SA, El-Naggar AK, Hanna EY (2012) Biology and management of salivary gland cancers. Semin Radiat Oncol 22:245–253. https://doi.org/10.1016/j.semradonc.2012.03.009

    Article  PubMed  Google Scholar 

  7. Dillon PM, Chakraborty S, Moskaluk CS, Joshi PJ, Thomas CY (2016) Adenoid cystic carcinoma: a review of recent advances, molecular targets, and clinical trials. Head Neck 38:620–627. https://doi.org/10.1002/hed.23925

    Article  PubMed  Google Scholar 

  8. Persson M, Andrén Y, Mark J, Horlings HM, Persson F, Stenman G (2009) Recurrent fusion of MYB and NFIB transcription factor genes in carcinomas of the breast and head and neck. Proc Natl Acad Sci U S A 106:18740–18744. https://doi.org/10.1073/pnas.0909114106

    Article  PubMed  PubMed Central  Google Scholar 

  9. Mitani Y, Liu B, Rao PH, Borra VJ, Zafereo M, Weber RS, Kies M, Lozano G, Futreal A, Caulin C, El-Naggar AK (2016) Novel MYBL1 gene rearrangements with recurrent MYBL1-NFIB fusions in salivary adenoid cystic carcinomas lacking t(6;9) translocations. Clin Cancer Res 22:725–733. https://doi.org/10.1158/1078-0432.CCR-15-2867-T

    Article  PubMed  CAS  Google Scholar 

  10. Ho AS, Kannan K, Roy DM et al (2013) The mutational landscape of adenoid cystic carcinoma. Nat Genet 45:791–798. https://doi.org/10.1038/ng.2643

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  11. Liu B, Mitani Y, Rao X, Zafereo M, Zhang J, Zhang J, Futreal A, Lozano G, El-Naggar AK (2017) Spatio-temporal genomic heterogeneity, phylogeny, and metastatic evolution in salivary adenoid cystic carcinoma. JNCI J Natl Cancer Inst. https://doi.org/10.1093/jnci/djx033

  12. Sajed DP, Faquin WC, Carey C, Severson EA, Afrogheh A, Johnson C, Blacklow SC, Chau NG, Lin DT, Krane JF, Jo VY, Garcia JJ, Scholl LM, Aster JC (2017) Diffuse staining for activated NOTCH1 correlates with NOTCH1 mutation status and is associated with worse outcome in adenoid cystic carcinoma. Am J Surg Pathol 41:1473–1482. https://doi.org/10.1097/PAS.0000000000000945

    Article  PubMed  Google Scholar 

  13. Ferrarotto R, Mitani Y, Diao L et al (2017) Activating NOTCH1 mutations define a distinct subgroup of patients with adenoid cystic carcinoma who have poor prognosis, propensity to bone and liver metastasis, and potential responsiveness to Notch1 inhibitors. J Clin Oncol 35:352–360. https://doi.org/10.1200/JCO.2016.67.5264

    Article  PubMed  CAS  Google Scholar 

  14. Frerich CA, Brayer KJ, Painter BM, Kang H, Mitani Y, El-naggar AK, Ness SA (2017) Transcriptomes define distinct subgroups of salivary gland adenoid cystic carcinoma with different driver mutations and outcomes. Oncotarget 9:7341–7358. https://doi.org/10.18632/oncotarget.23641

    Article  PubMed  PubMed Central  Google Scholar 

  15. Spiro RH (1997) Distant metastasis in adenoid cystic carcinoma of salivary origin. Am J Surg 174:495–498

    Article  PubMed  CAS  Google Scholar 

  16. Filipowicz W, Bhattacharyya SN, Sonenberg N (2008) Mechanisms of post-transcriptional regulation by microRNAs: are the answers in sight? Nat Rev Genet 9:102–114. https://doi.org/10.1038/nrg2290

    Article  PubMed  CAS  Google Scholar 

  17. Ul Hussain M (2012) Micro-RNAs (miRNAs): genomic organisation, biogenesis and mode of action. Cell Tissue Res 349:405–413. https://doi.org/10.1007/s00441-012-1438-0

    Article  PubMed  CAS  Google Scholar 

  18. Calin GA, Croce CM (2006) MicroRNA signatures in human cancers. Nat Rev Cancer 6:857–866. https://doi.org/10.1038/nrc1997

    Article  PubMed  CAS  Google Scholar 

  19. Thomson JM, Newman M, Parker JS, Morin-Kensicki EM, Wright T, Hammond SM (2006) Extensive post-transcriptional regulation of microRNAs and its implications for cancer. Genes Dev 20:22022207. https://doi.org/10.1101/gad.1444406

    Article  CAS  Google Scholar 

  20. Kong YW, Ferland-McCollough D, Jackson TJ, Bushell M (2012) MicroRNAs in cancer management. Lancet Oncol 13:e249–e258. https://doi.org/10.1016/S1470-2045(12)70073-6

    Article  PubMed  CAS  Google Scholar 

  21. Mitani Y, Roberts DB, Fatani H, Weber RS, Kies MS, Lippman SM, El-Naggar AK (2013) MicroRNA profiling of salivary adenoid cystic carcinoma: association of miR-17-92 upregulation with poor outcome. PLoS One 8:e66778. https://doi.org/10.1371/journal.pone.0066778

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  22. McShane LM, Altman DG, Sauerbrei W, Taube SE, Gion M, Clark GM (2005) REporting recommendations for tumour MARKer prognostic studies (REMARK). Br J Cancer 93:387–391. https://doi.org/10.1038/sj.bjc.6602678

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  23. Moore HM, Kelly AB, Jewell SD, McShane LM, Clark DP, Greenspan R, Hayes DF, Hainaut P, Kim P, Mansfield E, Potapova O, Riegman P, Rubinstein Y, Seijo E, Somiari S, Watson P, Weier HU, Zhu C, Vaught J (2011) Biospeciment reporting for improved study quality (BRISQ). J Proteome Res 10:3429–3438. https://doi.org/10.1021/pr200021n

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  24. Erichsen R, Lash TL, Hamilton-Dutoit SJ, Bjerregaard B, Vyberg M, Pedersen L (2010) Existing data sources for clinical epidemiology: the Danish National Pathology Registry and Data Bank. Clin Epidemiol 2:51–56

    Article  PubMed  PubMed Central  Google Scholar 

  25. Bjørndal K, Krogdahl A, Therkildsen MH, Overgaard J, Johansen J, Kristensen CA, Homøe P, Sørensen CH, Andersen E, Bundgaard T, Primdahl H, Lambertsen H, Andersen LJ, Godballe C (2011) Salivary gland carcinoma in Denmark 1990-2005: a national study of incidence, site and histology. Results of the Danish head and neck Cancer group (DAHANCA). Oral Oncol 47:677–682. https://doi.org/10.1016/j.oraloncology.2011.04.020

    Article  PubMed  Google Scholar 

  26. Stenman G, Licitra L, Said-Al-Naief N, van Zante A, Yarborough W (2017) Adenoid cystic carcinoma. In: El-Naggar A, Chan J, Grandis J, Takata T, Slootweg P (eds) World Health Organization classification of tumours. IARC Press, Lyon, 164–165

    Google Scholar 

  27. Andreasen S, Bjørndal K, Agander TK, Wessel I, Homøe P (2016) Tumors of the sublingual gland: a national clinicopathologic study of 29 cases. Eur Arch Otorhinolaryngol 273:3847–3856. https://doi.org/10.1007/s00405-016-4000-y

    Article  PubMed  Google Scholar 

  28. Andreasen S, Bishop JA, Hansen TV, Westra WH, Bilde A, von Buchwald C, Kiss K (2017) Human Papillomavirus-related Carcinoma with Adenoid Cystic-like Features of the Sinonasal Tract: Clinical and Morphological Characterization of 6 New Cases. Histopathology 70:880–888. https://doi.org/10.1111/his.13162

    Article  PubMed  Google Scholar 

  29. Bishop JA, Andreasen S, Hang JF et al (2017) HPV-related Multiphenotypic Sinonasal Carcinoma. Am J Surg Pathol 41:1690–1701. https://doi.org/10.1097/PAS.0000000000000944

    Article  PubMed  Google Scholar 

  30. Andreasen S, Melchior LC, Kiss K, Bishop JA, Høgdall E, Grauslund M, Wessel I, Homøe P, Agander TK (2018) The PRKD1 E710D hotspot mutation is highly specific in separating polymorphous adenocarcinoma of the palate from adenoid cystic carcinoma and pleomorphic adenoma on FNA. Cancer Cytopathol 126:275–281. https://doi.org/10.1002/cncy.21959

    Article  PubMed  CAS  Google Scholar 

  31. Pedersen CB (2011) The danish civil registration system. Scand J Public Health 39:22–25. https://doi.org/10.1177/1403494810387965

    Article  PubMed  Google Scholar 

  32. Helweg-Larsen K (2011) The danish register of causes of death. Scand J Public Health 39:26–29. https://doi.org/10.1177/1403494811399958

    Article  PubMed  Google Scholar 

  33. Howe K (2017) Extraction of miRNAs from formalin-fixed paraffin-embedded (FFPE) tissues. Methods Mol Biol. https://doi.org/10.1007/978-1-4939-6524-3_3

  34. Bolstad BM, Irizarry RA, Astrand M, Speed TP (2003) A comparison of normalization methods for high density oligonucleotide array data based on variance and bias. Bioinformatics 19:185–193

    Article  PubMed  CAS  Google Scholar 

  35. Skálová A, Vanecek T, Sima R, Laco J, Weinreb I, Perez-Ordonez B, Starek I, Geierova M, Simpson RH, Passador-Santos F, Ryska A, Leivo I, Kinkor Z, Michal M (2010) Mammary analogue secretory carcinoma of salivary glands, containing the ETV6-NTRK3 fusion gene: a hitherto undescribed salivary gland tumor entity. Am J Surg Pathol 34:599–608. https://doi.org/10.1097/PAS.0b013e3181d9efcc

    Article  PubMed  Google Scholar 

  36. Hudson JB, Collins BT (2014) MYB gene abnormalities t(6;9) in adenoid cystic carcinoma fine-needle aspiration biopsy using fluorescence in situ hybridization. Arch Pathol Lab Med 138:403–409. https://doi.org/10.5858/arpa.2012-0736-OA

    Article  PubMed  Google Scholar 

  37. Bass BP, Engel KB, Greytak SR, Moore HM (2014) A review of preanalytical factors affecting molecular, protein, and morphological analysis of formalin-fixed, paraffin-embedded (FFPE) tissue. How well do you know your FFPE specimen? Arch Pathol Lab Med 138:1520–1530. https://doi.org/10.5858/arpa.2013-0691-RA

    Article  PubMed  Google Scholar 

  38. Hall JS, Taylor J, Valentine HR, Irlam JJ, Eustace A, Hoskin PJ, Miller CJ, West CM (2012) Enhanced stability of microRNA expression facilitates classification of FFPE tumour samples exhibiting near total mRNA degradation. Br J Cancer 107:684–894. https://doi.org/10.1038/bjc.2012.294

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  39. Xi Y, Nakajima G, Gavin E, Morris CG, Kudo K, Hayashi K, Ju J (2007) Systematic analysis of microRNA expression of RNA extracted from fresh frozen and formalin-fixed paraffin-embedded samples. RNA 13:1668–1674. https://doi.org/10.1261/rna.642907

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  40. Yu SL, Chen HY, Chang GC et al (2008) MicroRNA signature predicts survival and relapse in lung cancer. Cancer Cell 13:48–57. https://doi.org/10.1016/j.ccr.2007.12.008

    Article  PubMed  CAS  Google Scholar 

  41. Spahn M, Kneitz S, Scholz CJ, Stenger N, Rüdiger T, Ströbel P, Riedmiller H, Kneitz B (2010) Expression of microRNA-221 is progressively reduced in aggressive prostate cancer and metastasis and predicts clinical recurrence. Int J Cancer 127:394–403. https://doi.org/10.1002/ijc.24715

    Article  PubMed  CAS  Google Scholar 

  42. Schetter AJ, Leung SY, Sohn JJ, Zanetti KA, Bowman ED, Yanaihara N, Yuen ST, Chan TL, Kwong DL, Au GK, Liu CG, Calin GA, Croce CM, Harris CC (2008) MicroRNA expression profiles associated with prognosis and therapeutic outcome in colon adenocarcinoma. JAMA 299:425–436. https://doi.org/10.1001/jama.299.4.425

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  43. Amit M, Binenbaum Y, Trejo–Leider L, Sharma K, Ramer N, Ramer I, Agbetoba A, Miles B, Xang X, Lei D, Bjørndal K, Godballe C, Mücke T, Wolff KD, Eckardt AM, Copelli C, Sesenna E, Palmer F, Ganly I, Patel S, Gil Z (2015) International collaborative validation of intraneural invasion as a prognostic marker in adenoid cystic carcinoma of the head and neck. Head Neck 37:1038–1045. https://doi.org/10.1002/hed.23710

    Article  PubMed  Google Scholar 

  44. Barrett AW, Speight PM (2009) Perineural invasion in adenoid cystic carcinoma of the salivary glands: a valid prognostic indicator? Oral Oncol 45:936–940. https://doi.org/10.1016/j.oraloncology.2009.07.001

    Article  PubMed  Google Scholar 

  45. Ma W, Kang Y, Ning L, Tan J, Wang H, Ying Y (2017) Identification of microRNAs involved in gefitinib resistance of non-small-cell lung cancer through the insulin-like growth factor receptor 1 signaling pathway. Exp Ther Med 14:2853–2862. https://doi.org/10.3892/etm.2017.4847

    Article  PubMed  PubMed Central  Google Scholar 

  46. Andersson MK, Afshari MK, Andre Y, Wick MJ, Stenman G (2017) Targeting the oncogenic transcriptional regulator MYB in adenoid cystic carcinoma by inhibition of IGF1R/AKT signaling. J Natl Cancer Inst. https://doi.org/10.1093/jnci/djx017

  47. Jang JS, Simon VA, Feddersen RM, Rakhshan F, Schultz DA, Zschunke MA, Lingle WL, Kolbert CP, Jen J (2011) Quantitative miRNA expression analysis using fluidigm microfluidics dynamic arrays. BMC Genomics 12:144. https://doi.org/10.1186/1471-2164-12-144

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  48. Chen Y, Gelfond JA, McManus LM, Shireman PK (2009) Reproducibility of quantitative RT-PCR array in miRNA expression profiling and comparison with microarray analysis. BMC Genomics 10:407. https://doi.org/10.1186/1471-2164-10-407

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  49. Mestdagh P, Feys T, Bernard N, Guenther S, Chen C, Speleman F, Vandesomplete J (2008) High-throughput stem-loop RT-qPCR miRNA expression profiling using minute amounts of input RNA. Nucleic Acids Res 36:e143. https://doi.org/10.1093/nar/gkn725

    Article  PubMed  PubMed Central  CAS  Google Scholar 

  50. Pérez-Rivas LG, Jerez JM, Carmona R, de Luque V, Vicioso L, Claros MG, Viguera E, Pajares B, Sánchez A, Ribelles N, Alba E, Lozano J (2014) A microRNA signature associated with early recurrence in breast cancer. PLoS One 14:e91884. https://doi.org/10.1371/journal.pone.0091884

    Article  CAS  Google Scholar 

Download references

Acknowledgements

We wish to thank Katarina Larsen and Lene Pedersen for valuable assistance with miRNA extraction.

Funding

SA was supported by Einar Willumsen Foundation, Merchant Kristjan Kjær and wife Margrethe Kjær’s Foundation, The Danielsen Foundation, Else and Mogens Wedell-Wedellsborg’s Foundation, Region Zealand’s Research Fund, Hans Skouby and wife Emma Skouby’s Foundation, and the A.P. Møller Foundation for the Advancement of Medical Research.

Author information

Authors and Affiliations

  1. Department of Otorhinolaryngology and Maxillofacial Surgery, Zealand University Hospital, Køge, Denmark

    Simon Andreasen & Preben Homøe

  2. Department of Otorhinolaryngology Head and Neck Surgery and Audiology, Rigshospitalet, Copenhagen, Denmark

    Simon Andreasen & Irene Wessel

  3. Unit of Human Genetics, Department of Clinical Research, University of Southern Denmark, Odense, Denmark

    Qihua Tan

  4. Department of Pathology, Rigshospitalet, Copenhagen, Denmark

    Tina Klitmøller Agander & Steffen Heegaard

  5. Genomic Medicine, Rigshospitalet, Copenhagen University Hospital, Copenhagen, Denmark

    Thomas V. O. Hansen

  6. Department of Pathology, Charles University in Prague, Faculty of Medicine, Pilsen, Czech Republic

    Petr Steiner

  7. Bioptic Laboratory Ltd, Molecular Pathology Laboratory, Pilsen, Czech Republic

    Petr Steiner

  8. Department of ORL–Head and Neck Surgery, Odense University Hospital, Odense, Denmark

    Kristine Bjørndal

  9. Department of Pathology, Herlev Hospital, University of Copenhagen, Herlev, Denmark

    Estrid Høgdall

  10. Department of Pathology, Odense University Hospital, Odense, Denmark

    Stine Rosenkilde Larsen

  11. Department of Pathology, Aalborg University Hospital, Aalborg, Denmark

    Daiva Erentaite

  12. Department of Pathology, Zealand University Hospital, Roskilde, Denmark

    Caroline Holkmann Olsen

  13. Department of Pathology, Aarhus University Hospital, Aarhus, Denmark

    Benedicte Parm Ulhøi

  14. Department of Ophthalmology, Rigshospitalet-Glostrup, Copenhagen, Denmark

    Steffen Heegaard

Authors
  1. Simon Andreasen
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Qihua Tan
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Tina Klitmøller Agander
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Thomas V. O. Hansen
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Petr Steiner
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Kristine Bjørndal
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Estrid Høgdall
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Stine Rosenkilde Larsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Daiva Erentaite
    View author publications

    You can also search for this author in PubMed Google Scholar

  10. Caroline Holkmann Olsen
    View author publications

    You can also search for this author in PubMed Google Scholar

  11. Benedicte Parm Ulhøi
    View author publications

    You can also search for this author in PubMed Google Scholar

  12. Steffen Heegaard
    View author publications

    You can also search for this author in PubMed Google Scholar

  13. Irene Wessel
    View author publications

    You can also search for this author in PubMed Google Scholar

  14. Preben Homøe
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Simon Andreasen.

Ethics declarations

Conflicts of interest

The authors declare that they have no conflict of interest.

Electronic supplementary material

ESM 1

(DOCX 134 kb)

Table S1

(DOCX 18 kb)

Table S2

(DOCX 14 kb)

Table S3

(DOCX 43 kb)

Table S4

(XLSX 3328 kb)

Table S5

(DOCX 119 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Andreasen, S., Tan, Q., Agander, T.K. et al. MicroRNA dysregulation in adenoid cystic carcinoma of the salivary gland in relation to prognosis and gene fusion status: a cohort study. Virchows Arch 473, 329–340 (2018). https://doi.org/10.1007/s00428-018-2423-0

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s00428-018-2423-0

Keywords

Search

Navigation