Virchows Archiv

, Volume 458, Issue 5, pp 583–592 | Cite as

Aberrant PLAG1 expression in pleomorphic adenomas of the salivary gland: a molecular genetic and immunohistochemical study

  • Atsuji MatsuyamaEmail author
  • Masanori Hisaoka
  • Yuichi Nagao
  • Hiroshi Hashimoto
Original Article


The morphologic distinction of pleomorphic adenoma from other benign or low-grade salivary gland tumors is sometimes difficult and problematic because of their potentially overlapping histological patterns. A subset of pleomorphic adenoma harbors specific gene alterations involving PLAG1 or HMGA2, and the detection of these fusion genes and their products using formalin-fixed, paraffin-embedded (FFPE) tumor specimens may be a useful diagnostic adjunct. In the present study, gene fusions involving PLAG1 or HMGA2 were examined by reverse transcription-polymerase chain reaction (RT-PCR) analysis, with FFPE tumor tissues and immunohistochemical expression of PLAG1 in 45 pleomorphic adenomas, using a commercially available antibody. RT-PCR analyses identified the CTNNB1-PLAG1, LIFR-PLAG1, CHCHD7-PLAG1, and HMGA2-WIF1 fusion transcripts in eight, two, one, and one case, respectively. The TCEA1-PLAG1, HMGA2-FHIT, and HMGA2-NFIB fusion transcripts were not detected. Immunohistochemically, tumor cells in all 45 pleomorphic adenomas were positive for PLAG1, irrespective of PLAG1 rearrangements, even in the case with the HMGA2-WIF1 fusion transcript. Tumor cells displaying myoepithelial or cartilaginous differentiation were almost constantly positive for PLAG1, whereas a limited expression was observed in glandular or keratinizing cells. Among the 46 tumors other than pleomorphic adenoma, 4 carcinomatous components of carcinomas ex pleomorphic adenoma were positive for PLAG1, the other 39 were negative for PLAG1, and the remaining 3 were only faintly and/or focally stained, indicating that the immunohistochemical detection of PLAG1 is diagnostically useful. The present results also suggest that overexpression of PLAG1 is essential for the tumorigenesis of pleomorphic adenomas, although the mechanisms mediating PLAG1 overexpression seem to be variable.


Pleomorphic adenoma PLAG1 HMGA2 Immunohistochemistry RT-PCR 



The authors thank Kazumi Tanaka for her technical assistance.

Conflict of interest

We declare that we have no conflict of interest.


  1. 1.
    Barnes L, Eveson JW, Reichart PA, Sidransky D (eds) (2005) World Health Organization classification of tumours. Pathology and genetics of head and neck tumours. IARC Press, LyonGoogle Scholar
  2. 2.
    Asp J, Persson F, Kost-Alimova M, Stenman G (2006) CHCHD7-PLAG1 and TCEA1-PLAG1 gene fusions resulting from cryptic, intrachromosomal 8q rearrangements in pleomorphic salivary gland adenomas. Genes Chromosom Cancer 45:820–828PubMedCrossRefGoogle Scholar
  3. 3.
    Aström AK, Voz ML, Kas K, Röijer E, Wedell B, Mandahl N, Van de Ven W, Mark J, Stenman G (1999) Conserved mechanism of PLAG1 activation in salivary gland tumors with and without chromosome 8q12 abnormalities: identification of SII as a new fusion partner gene. Cancer Res 59:918–923PubMedGoogle Scholar
  4. 4.
    Martins C, Fonseca I, Roque L, Pereira T, Ribeiro C, Bullerdiek J, Soares J (2005) PLAG1 gene alterations in salivary gland pleomorphic adenoma and carcinoma ex-pleomorphic adenoma: a combined study using chromosome banding, in situ hybridization and immunocytochemistry. Mod Pathol 18:1048–1055PubMedCrossRefGoogle Scholar
  5. 5.
    Kas K, Voz ML, Röijer E, Aström AK, Meyen E, Stenman G, Van de Ven WJ (1997) Promoter swapping between the genes for a novel zinc finger protein and beta-catenin in pleiomorphic adenomas with t(3;8)(p21;q12) translocations. Nat Genet 15:170–174PubMedCrossRefGoogle Scholar
  6. 6.
    Voz ML, Aström AK, Kas K, Mark J, Stenman G, Van de Ven WJ (1998) The recurrent translocation t(5;8)(p13;q12) in pleomorphic adenomas results in upregulation of PLAG1 gene expression under control of the LIFR promoter. Oncogene 16:1409–1416PubMedCrossRefGoogle Scholar
  7. 7.
    Bullerdiek J, Wobst G, Meyer-Bolte K, Chilla R, Haubrich J, Thode B, Bartnitzke S (1993) Cytogenetic subtyping of 220 salivary gland pleomorphic adenomas: correlation to occurrence, histological subtype, and in vitro cellular behavior. Cancer Genet Cytogenet 65:27–31PubMedCrossRefGoogle Scholar
  8. 8.
    Mark J, Dahlenfors R, Wedell B (1997) Impact of the in vitro technique used on the cytogenetic patterns in pleomorphic adenomas. Cancer Genet Cytogenet 95:9–15PubMedCrossRefGoogle Scholar
  9. 9.
    Geurts JM, Schoenmakers EF, Röijer E, Aström AK, Stenman G, van de Ven WJ (1998) Identification of NFIB as recurrent translocation partner gene of HMGIC in pleomorphic adenomas. Oncogene 16:865–872PubMedCrossRefGoogle Scholar
  10. 10.
    Geurts JM, Schoenmakers EF, Röijer E, Stenman G, Van de Ven WJ (1997) Expression of reciprocal hybrid transcripts of HMGIC and FHIT in a pleomorphic adenoma of the parotid gland. Cancer Res 57:13–17PubMedGoogle Scholar
  11. 11.
    Persson F, Andrén Y, Winnes M, Wedell B, Nordkvist A, Gudnadottir G, Dahlenfors R, Sjögren H, Mark J, Stenman G (2009) High-resolution genomic profiling of adenomas and carcinomas of the salivary glands reveals amplification, rearrangement, and fusion of HMGA2. Genes Chromosom Cancer 48:69–82PubMedCrossRefGoogle Scholar
  12. 12.
    Hisaoka M, Tsuji S, Morimitsu Y, Hashimoto H, Shimajiri S, Komiya S, Ushijima M (1998) Detection of TLS/FUS-CHOP fusion transcripts in myxoid and round cell liposarcomas by nested reverse transcription-polymerase chain reaction using archival paraffin-embedded tissues. Diagn Mol Pathol 7:96–101PubMedCrossRefGoogle Scholar
  13. 13.
    Tsuji S, Hisaoka M, Morimitsu Y, Hashimoto H, Shimajiri S, Komiya S, Ushijima M, Nakamura T (1998) Detection of SYT-SSX fusion transcripts in synovial sarcoma by reverse transcription-polymerase chain reaction using archival paraffin-embedded tissues. Am J Pathol 153:1807–1812PubMedCrossRefGoogle Scholar
  14. 14.
    Argani P, Zakowski MF, Klimstra DS, Rosai J, Ladanyi M (1998) Detection of the SYT-SSX chimeric RNA of synovial sarcoma in paraffin-embedded tissue and its application in problematic cases. Mod Pathol 11:65–71PubMedGoogle Scholar
  15. 15.
    Li XQ, Hisaoka M, Shi DR, Zhu XZ, Hashimoto H (2004) Expression of anaplastic lymphoma kinase in soft tissue tumors: an immunohistochemical and molecular study of 249 cases. Hum Pathol 35:711–721PubMedCrossRefGoogle Scholar
  16. 16.
    Okabe M, Miyabe S, Nagatsuka H, Terada A, Hanai N, Yokoi M, Shimozato K, Eimoto T, Nakamura S, Nagai N, Hasegawa Y, Inagaki H (2006) MECT1-MAML2 fusion transcript defines a favorable subset of mucoepidermoid carcinoma. Clin Cancer Res 12:3902–3907PubMedCrossRefGoogle Scholar
  17. 17.
    Llombart-Bosch A, Machado I, Navarro S, Bertoni F, Bacchini P, Alberghini M, Karzeladze A, Savelov N, Petrov S, Alvarado-Cabrero I, Mihaila D, Terrier P, Lopez-Guerrero JA, Picci P (2009) Histological heterogeneity of Ewing's sarcoma/PNET: an immunohistochemical analysis of 415 genetically confirmed cases with clinical support. Virchows Arch 455:397–411PubMedCrossRefGoogle Scholar
  18. 18.
    Barnoud R, Sabourin JC, Pasquier D, Ranchère D, Bailly C, Terrier-Lacombe MJ, Pasquier B (2000) Immunohistochemical expression of WT1 by desmoplastic small round cell tumor: a comparative study with other small round cell tumors. Am J Surg Pathol 24:830–836PubMedCrossRefGoogle Scholar
  19. 19.
    Argani P, Lal P, Hutchinson B, Lui MY, Reuter VE, Ladanyi M (2003) Aberrant nuclear immunoreactivity for TFE3 in neoplasms with TFE3 gene fusions: a sensitive and specific immunohistochemical assay. Am J Surg Pathol 27:750–761PubMedCrossRefGoogle Scholar
  20. 20.
    Nakamura S, Shiota M, Nakagawa A, Yatabe Y, Kojima M, Motoori T, Suzuki R, Kagami Y, Ogura M, Morishima Y, Mizoguchi Y, Okamoto M, Seto M, Koshikawa T, Mori S, Suchi T (1997) Anaplastic large cell lymphoma: a distinct molecular pathologic entity: a reappraisal with special reference to p80(NPM/ALK) expression. Am J Surg Pathol 21:1420–1432PubMedCrossRefGoogle Scholar
  21. 21.
    Takeuchi K, Choi YL, Soda M, Inamura K, Togashi Y, Hatano S, Enomoto M, Takada S, Yamashita Y, Satoh Y, Okumura S, Nakagawa K, Ishikawa Y, Mano H (2008) Multiplex reverse transcription-PCR screening for EML4-ALK fusion transcripts. Clin Cancer Res 14:6618–6624PubMedCrossRefGoogle Scholar
  22. 22.
    Tomlins SA, Rhodes DR, Perner S, Dhanasekaran SM, Mehra R, Sun XW, Varambally S, Cao X, Tchinda J, Kuefer R, Lee C, Montie JE, Shah RB, Pienta KJ, Rubin MA, Chinnaiyan AM (2005) Recurrent fusion of TMPRSS2 and ETS transcription factor genes in prostate cancer. Science 310:644–648PubMedCrossRefGoogle Scholar
  23. 23.
    Sandros J, Stenman G, Mark J (1990) Cytogenetic and molecular observations in human and experimental salivary gland tumors. Cancer Genet Cytogenet 44:153–167PubMedCrossRefGoogle Scholar
  24. 24.
    Rohen C, Rogalla P, Meyer-Bolte K, Bartnitzke S, Chilla R, Bullerdiek J (1999) Pleomorphic adenomas of the salivary glands: absence of HMGIY rearrangements. Cancer Genet Cytogenet 111:178–181PubMedCrossRefGoogle Scholar
  25. 25.
    Jin C, Martins C, Jin Y, Wiegant J, Wennerberg J, Dictor M, Gisselsson D, Strömbeck B, Fonseca I, Mitelman F, Tanke HJ, Höglund M, Mertens F (2001) Characterization of chromosome aberrations in salivary gland tumors by FISH, including multicolor COBRA-FISH. Genes Chromosom Cancer 30:161–167PubMedCrossRefGoogle Scholar
  26. 26.
    Debiec-Rychter M, Van Valckenborgh I, Van den Broeck C, Hagemeijer A, Van deVen WJ, Kas K, Van Damme B, Voz ML (2001) Histologic localization of PLAG1 (pleomorphic adenoma gene 1) in pleomorphic adenoma of the salivary gland: cytogenetic evidence of common origin of phenotypically diverse cells. Lab Invest 81:1289–1297PubMedCrossRefGoogle Scholar
  27. 27.
    el-Naggar AK, Lovell M, Callender DL, Ordonez NG, Killary AM (1999) Cytogenetic analysis of a primary salivary gland myoepithelioma. Cancer Genet Cytogenet 113:49–53PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2011

Authors and Affiliations

  • Atsuji Matsuyama
    • 1
    Email author
  • Masanori Hisaoka
    • 1
  • Yuichi Nagao
    • 1
  • Hiroshi Hashimoto
    • 1
  1. 1.Department of Pathology and Oncology, School of MedicineUniversity of Occupational and Environmental HealthKitakyushuJapan

Personalised recommendations