Advertisement

Breast Cancer Research and Treatment

, Volume 152, Issue 2, pp 305–312 | Cite as

Mutational analysis of MED12 in fibroadenomas and phyllodes tumors of the breast by means of targeted next-generation sequencing

  • Chieko Mishima
  • Naofumi Kagara
  • Tomonori Tanei
  • Yasuto Naoi
  • Masafumi Shimoda
  • Atsushi Shimomura
  • Kenzo Shimazu
  • Seung Jin Kim
  • Shinzaburo Noguchi
Preclinical study

Abstract

We aimed to analyze MED12 mutation in fibroadenomas (FAs) and phyllodes tumors (PTs) of the breast, which are closely related and consist of epithelial and stromal components. Targeted deep-sequencing using next-generation sequencing was performed in FAs (n = 58) and PTs (n = 27). The frequency of MED12 mutant tumors was significantly higher (P = 0.016) in PTs (74.1 %) than in FAs. (46.6 %). As for FAs, this frequency was significantly higher (P = 0.001) for intracanalicular type (69.0 %) than for other histological subtypes such as pericanalicular, organoid, and mastopathic types (24.1 %). Laser microdissection study revealed that stromal cells, but not epithelial cells, harbored MED12 mutations in both FAs and PTs. MED12 mutation is implicated in the pathogenesis of both FAs and PTs. The similarly high frequency of MED12 mutation in intracanalicular type FAs suggests that they are most closely related to PTs. It is thus speculated that FAs with MED12 mutation are more likely to progress to PTs.

Keywords

MED12 Somatic mutation Next-generation sequencing Fibroepithelial tumor 

Abbreviations

FA

Fibroadenoma

PT

Phyllodes tumor

Notes

Acknowledgments

We are grateful to Dr. Junichiro Ikeda for his help in the pathological diagnosis.

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical standards

The study complied with the current laws of Japan.

Supplementary material

10549_2015_3469_MOESM1_ESM.tif (75 kb)
Supplementary material 1 (TIFF 75 kb). Primers for next-generation sequencing of MED12. Two sets of primers were designed to amplify genomic DNA (gDNA) and complementary DNA (cDNA). Letters in bold type: primer sequences
10549_2015_3469_MOESM2_ESM.tif (233 kb)
Supplementary material 2 (TIFF 233 kb). MED12 mutation status of thirteen synchronous multiple tumors from six patients (a), and seven metachronous multiple tumors from three patients (b). Filled circle, fibroadenoma; circle with vertical lines, phyllodes tumor. Period after the resection of primary tumor (*); period after the resection of secondary tumor (**)
10549_2015_3469_MOESM3_ESM.doc (30 kb)
Supplementary material 3 (DOC 30 kb)

References

  1. 1.
    Carty NJ, Carter C, Rubin C, Ravichandran D, Royle GT, Taylor I (1995) Management of fibroadenoma of the breast. Ann R Coll Surg Engl 77:127–130PubMedCentralPubMedGoogle Scholar
  2. 2.
    Dixon JM (1991) Cystic disease and fibroadenoma of the breast: natural history and relation to breast cancer risk. Br Med Bull 47:258–271PubMedGoogle Scholar
  3. 3.
    Noguchi S, Yokouchi H, Aihara T, Motomura K, Inaji H, lmaoka S, Koyama H (1995) Progression of fibroadenoma to phyllodes tumor demonstrated by clonal analysis. Cancer 76:1779–1785PubMedCrossRefGoogle Scholar
  4. 4.
    Kuijper A (2006) Pathogenesis and progression of fibroepithelial breast tumors. Utrecht University Repository, NederlandGoogle Scholar
  5. 5.
    Yang S, Kandil D, Cosar EF, Khan A (2014) Fibroepithelial tumors of the breast pathologic and immunohistochemical features and molecular mechanisms. Arch Pathol Lab Med 138:25–36PubMedCrossRefGoogle Scholar
  6. 6.
    Abe M, Miyata S, Nishimura S, Iijima K, Makita M, Akiyama F, Iwase T (2011) Malignant transformation of breast fibroadenoma to malignant phyllodes tumor: long-term outcome of 36 malignant phyllodes tumors. Breast Cancer 18:268–272PubMedCrossRefGoogle Scholar
  7. 7.
    Kuijper A, Buerger H, Simon R, Schaefer K, Croonen A, Boecker Q, Wall E, Diest PJ (2002) Analysis of the progression of fibroepithelial tumours of the breast by PCR-based clonality assay. J Pathol 197:575–581PubMedCrossRefGoogle Scholar
  8. 8.
    Noguchi S, Aihara T, Motomura K, Inaji H, Imaoka S, Koyama H, Kasugai T, Wada A (1996) Phyllodes tumor of the breast: pathology, genesis, diagnosis, and treatment. Breast Cancer 3:79–92PubMedCrossRefGoogle Scholar
  9. 9.
    Lim WK, Ong CK, Tan J, Thike AA, Ng CC, Rajasegaran V, Myint SS, Nagarajan S, Nasir ND, McPherson JR, Cutcutache I, Poore G, Tay ST, Ooi WS, Tan VK, Hartman M, Ong KW, Tan BK, Rozen SG, Tan PH, Tan P, Teh BT (2014) Exome sequencing identifies highly recurrent MED12 somatic mutations in breast fibroadenoma. Nat Genet 46:877–880PubMedCrossRefGoogle Scholar
  10. 10.
    Mäkinen N, Mehine M, Tolvanen J, Kaasinen E, Li Y, Lehtonen HJ, Gentile M, Yan J, Enge M, Taipale M, Aavikko M, Katainen R, Virolainen E, Böhling T, Koski TA, Launonen V, Sjöberg J, Taipale J, Vahteristo P, Aaltonen LA (2011) MED12, the mediator complex subunit 12 gene, is mutated at high frequency in uterine leiomyomas. Science 334:252–255PubMedCrossRefGoogle Scholar
  11. 11.
    Taatjes DJ (2010) The human mediator complex: a versatile, genome-wide regulator of transcription. Trends Biochem Sci 35:315–322PubMedCentralPubMedCrossRefGoogle Scholar
  12. 12.
    Graham JM Jr, Schwartz CE (2013) MED12 related disorders. Am J Med Genet 161:2734–2740CrossRefGoogle Scholar
  13. 13.
    Turunen M, Spaeth JM, Keskitalo S, Park MJ, Kivioja T, Clark AD, Mäkinen N, Gao F, Palin K, Nurkkala H, Vähärautio A, Aavikko M, Kämpjärvi K, Vahteristo P, Kim CA, Aaltonen LA, Varjosalo M, Taipale J, Boyer TG (2014) Uterine leiomyoma-linked MED12 mutations disrupt mediator-associated CDK activity. Cell Reports 7:654–660PubMedCentralPubMedCrossRefGoogle Scholar
  14. 14.
    Cani AK, Hovelson DH, McDaniel AS, Sadis S, Haller MJ, Yadati V, Amin AM, Bratley J, Bandla S, Williams PD, Rhodes K, Liu CJ, Quist MJ, Rhodes DR, Grasso CS, Kleer CG, Tomlins SA (2015) Next-Gen sequencing exposes frequent MED12 mutations and actionable therapeutic targets in phyllodes tumors. Mol Cancer Res 13:613–619PubMedCrossRefGoogle Scholar
  15. 15.
    Yoshida M, Sekine S, Ogawa R, Yoshida H, Maeshima A, Kanai Y, Kinoshita T, Ochiai A (2015) Frequent MED12 mutations in phyllodes tumours of the breast. Br J Cancer 112:1703–1708PubMedCrossRefGoogle Scholar
  16. 16.
    Nagasawa S, Maeda I, Fukuda T, Wu W, Hayami R, Kojima Y, Tsugawa K, Ohta T (2015) MED12 exon 2 mutations in phyllodes tumors of the breast. Cancer Med. doi: 10.1002/cam4.462 PubMedGoogle Scholar
  17. 17.
    Piscuoglio S, Murray M, Fusco N, Marchiò C, Loo FL, Martelotto LG, Schultheis AM, Akram M, Weigelt B, Brogi E, Reis-Filho JS (2015) MED12 somatic mutations in fibroadenomas and phyllodes tumors of the breast. Histopathology. doi: 10.1111/his.12712 Google Scholar
  18. 18.
    Pfarr N, Kriegsmann M, Sinn P, Klauschen F, Endris V, Herpel E, Muckenhuber A, Jesinghaus M, Klosterhalfen B, Penzel R, Lennerz JK, Weichert W, Stenzinger A (2015) Distribution of MED12 mutations in fibroadenomas and phyllodes tumors of the breast-implications for tumor biology and pathological diagnosis. Genes Chromosom Cancer 54:444–452PubMedCrossRefGoogle Scholar
  19. 19.
    Kuroda H, Takeuchi I, Ohnishi K, Sakamoto G, Akiyama F, Toyozumi Y, Momose S, Tamaru J, Itoyama S (2006) Clinical and pathologic features of fibroadenoma of the mastopathic type. Surg Today 36:590–595PubMedCrossRefGoogle Scholar
  20. 20.
    Mori I, Han B, Wang X, Taniguchi E, Nakamura M, Nakamura Y, Bai Y, Kakudo K (2006) Mastopathic fibroadenoma of the breast: a pitfall of aspiration cytology. Cytopathology 17:233–238PubMedCrossRefGoogle Scholar
  21. 21.
    Feakins RM, Mulcahy HE, Nickols CD, Wells CA (1999) p53 expression in phyllodes tumours is associated with histological features of malignancy but does not predict outcome. Histopathology 35:162–169PubMedCrossRefGoogle Scholar
  22. 22.
    Tan PH, Jayabaskar T, Yip G, Tan Y, Hilmy M, Selvarajan S, Bay BH (2005) p53 and c-kit (CD117) protein expression as prognostic indicators in breast phyllodes tumors: a tissue microarray study. Mod Pathol 18:1527–1534PubMedCrossRefGoogle Scholar
  23. 23.
    Yonemori K, Hasegawa T, Shimizu C, Shibata T, Matsumoto K, Kouno T, Ando M, Katsumata N, Fujiwara Y (2006) Correlation of p53 and MIB-1 expression with both the systemic recurrence and survival in cases of phyllodes tumors of the breast. Pathol Res Pract 202:705–712PubMedCrossRefGoogle Scholar
  24. 24.
    Kleer CG, Giordano TJ, Braun T, Oberman HA (2001) Pathologic, immunohistochemical, and molecular features of benign and malignant phyllodes tumors of the breast. Mod Pathol 14:185–190PubMedCrossRefGoogle Scholar
  25. 25.
    Sawyer EJ, Poulsom R, Hunt FT, Jeffery R, Elia G, Ellis IO, Ellis P, Tomlinson IP, Hanby AM (2003) Malignant phyllodes tumours show stromal overexpression of c-myc and c-kit. J Pathol 200:59–64PubMedCrossRefGoogle Scholar
  26. 26.
    Giri D (2009) Recurrent challenges in the evaluation of fibroepithelial lesions. Arch Pathol Lab Med 133:713–721PubMedGoogle Scholar
  27. 27.
    Gatalica Z, Finkelstein S, Lucio E, Tawfik O, Palazzo J, Hightower B, Eyzaguirre E (2001) p53 protein expression and gene mutation in phyllodes tumors of the breast. Pathol Res Pract 197:183–187PubMedCrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Chieko Mishima
    • 1
  • Naofumi Kagara
    • 1
  • Tomonori Tanei
    • 1
  • Yasuto Naoi
    • 1
  • Masafumi Shimoda
    • 1
  • Atsushi Shimomura
    • 1
  • Kenzo Shimazu
    • 1
  • Seung Jin Kim
    • 1
  • Shinzaburo Noguchi
    • 1
  1. 1.Department of Breast and Endocrine Surgery, Faculty of MedicineOsaka University, Graduate School of MedicineSuita CityJapan

Personalised recommendations