Kallikrein-Related Peptidase mRNA Expression in Adenoid Cystic Carcinoma of Salivary Glands: A Polymerase Chain Reaction Study

  • Zachary Kerr
  • Alyssa Hayter
  • Zia Khan
  • Mark DarlingEmail author
Original Paper


Kallikrein-related peptidases (KLKs) are a group of 15 serine proteases implicated in a variety of biological processes. Aberrant expression of KLKs has been associated with the development of certain cancers. However, the role of KLKs in salivary tumors has not been extensively studied. This study evaluated the expression of KLKs in both adenoid cystic carcinoma (ACC) and normal salivary gland tissue. We isolated total RNA from 39 formalin-fixed, paraffin-embedded samples, which included 24 ACCs and 15 normal salivary gland tissues. Complementary DNA, synthesized by reverse transcription, was combined with gene specific kallikrein primers (KLK1KLK15) to allow for quantitative real-time PCR. Data was normalized to a β-actin housekeeping gene. Relative quantification analysis was performed using the ΔCq method. KLK1KLK15 expression was observed in both tissue types. However, KLK1, KLK8, KLK11, and KLK14 were found to be downregulated in ACC. We propose that this may represent a multi-parametric panel providing diagnostic and prognostic information.


Kallikreins Serine protease Adenoid cystic carcinoma RT-PCR Gene expression 



This study was partially funded by the Canadian Association of Oral and Maxillofacial Surgeons.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical Approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the Western University Research Ethics Board, Study Number 103783 and with the 1964 Helsinki declaration and its later amendments or comparable ethical standards. This article does not contain any studies with animals performed by any of the authors.


  1. 1.
    Ellis GL, Auclair PL. Tumors of the salivary glands. Series 4. In: Ellis GL, Auclair PL, editors. Washington DC: American Registry of Pathology; 2008.Google Scholar
  2. 2.
    El-Naggar AK, Chan JKC, Grandis JR, Takashi T, Slootweg PJ. WHO classification of head and neck tumours. 4th ed. In: El-Naggar AK, Chan JKC, Grandis JR, Takata TSP, editors; 2017.Google Scholar
  3. 3.
    Kokemueller H, Eckardt A, Brachvogel P, Hausamen J-E. Adenoid cystic carcinoma of the head and neck: a 20 years experience. Int J Oral Maxillofac Surg. 2004;33:25–31.CrossRefGoogle Scholar
  4. 4.
    Kokemueller H, Swennen G, Brueggemann N, Brachvogel P, Eckardt A, Hausamen J-E. Epithelial malignancies of the salivary glands: clinical experience of a single institution: a review. Int J Oral Maxillofac Surg. 2004;33:423–32.CrossRefGoogle Scholar
  5. 5.
    Kim KH, Sung MW, Chung PS, Rhee CS, Park CI, Kim WH. Adenoid cystic carcinoma of the head and neck. Arch Otolaryngol Head Neck Surg. 1994;120:721–6.CrossRefGoogle Scholar
  6. 6.
    Adelstein DJ, Koyfman SA, El-Naggar AK, Hanna EY. Biology and management of salivary gland cancers. Semin Radiat Oncol. 2012;22:245–53.CrossRefGoogle Scholar
  7. 7.
    Sur RK, Donde B, Levin V, Pacella J, Kotzen J, Cooper K, et al. Adenoid cystic carcinoma of the salivary glands: a review of 10 years. Laryngoscope. 1997;107:1276–80.CrossRefGoogle Scholar
  8. 8.
    de Almeida-Pinto YD, dos Costa SF, de Andrade BAB, Altemani A, Vargas PA, Abreu LG, et al. t(6;9)(MYB-NFIB) in head and neck adenoid cystic carcinoma: a systematic review with meta-analysis. Oral Dis. 2019;25:1277–82.CrossRefGoogle Scholar
  9. 9.
    Fujii K, Murase T, Beppu S, Saida K, Takino H, Masaki A, et al. MYB, MYBL1, MYBL2 and NFIB gene alterations and MYC overexpression in salivary gland adenoid cystic carcinoma. Histopathology. 2017;71:823–34.CrossRefGoogle Scholar
  10. 10.
    Shaw JLV, Diamandis EP. Distribution of 15 human kallikreins in tissues and biological fluids. Clin Chem. 2007;53:1423–32.CrossRefGoogle Scholar
  11. 11.
    Kalinska M, Meyer-Hoffert U, Kantyka T, Potempa J. Kallikreins: the melting pot of activity and function. Biochimie. 2016;122:270–82.CrossRefGoogle Scholar
  12. 12.
    Hollenberg MD. KLKs and their hormone-like signaling actions: a new life for the PSA-KLK family. Biol Chem. 2014;395:915–29.CrossRefGoogle Scholar
  13. 13.
    Kontos CK, Scorilas A. Kallikrein-related peptidases (KLKs): a gene family novel cancer biomarkers. Clin Chem Lab Med. 2012;50:1877–91.CrossRefGoogle Scholar
  14. 14.
    Kryza T, Silva ML, Loessner D, Heuzé-Vourc’h N, Clements JA. The kallikrein-related peptidase family: dysregulation and functions during cancer progression. Biochimie. 2016;122:283–99.CrossRefGoogle Scholar
  15. 15.
    Lawrence MG, Lai J, Clements JA. Kallikreins on steroids: structure, function, and hormonal regulation of prostate-specific antigen and the extended kallikrein locus. Endocr Rev. 2010;31:407–46.CrossRefGoogle Scholar
  16. 16.
    Dong Y, Loessner D, Irving-Rodgers H, Obermair A, Nicklin JL, Clements JA. Metastasis of ovarian cancer is mediated by kallikrein related peptidases. Clin Exp Metastasis. 2014;31:135–47.CrossRefGoogle Scholar
  17. 17.
    Prassas I, Eissa A, Poda G, Diamandis EP. Unleashing the therapeutic potential of human kallikrein-related serine proteases. Nat Rev Drug Discov. 2015;14:183–202.CrossRefGoogle Scholar
  18. 18.
    Darling MR, Tsai S, Jackson-Boeters L, Daley TD, Diamandis EP. Human kallikrein 8 expression in salivary gland tumors. Head Neck Pathol. 2008;2(3):169–74.CrossRefGoogle Scholar
  19. 19.
    Borgoño CA, Diamandis EP. The emerging roles of human tissue kallikreins in cancer. Nat Rev Cancer. 2004;4:876–90.CrossRefGoogle Scholar
  20. 20.
    Sher Y-P, Chou C-C, Chou R-H, Wu H-M, Wayne Chang W-S, Chen C-H, et al. Human kallikrein 8 protease confers a favorable clinical outcome in non-small cell lung cancer by suppressing tumor cell invasiveness. Cancer Res. 2006;66:11763–70.CrossRefGoogle Scholar
  21. 21.
    Magklara A, Scorilas A, Katsaros D, Massobrio M, Yousef GM, Fracchioli S, et al. The human KLK8 (neuropsin/ovasin) gene: identification of two novel splice variants and its prognostic value in ovarian cancer. Clin Cancer Res. 2001;7:806–11.Google Scholar
  22. 22.
    Borgoño CA, Kishi T, Scorilas A, Harbeck N, Dorn J, Schmalfeldt B, et al. Human kallikrein 8 protein is a favorable prognostic marker in ovarian cancer. Clin Cancer Res. 2008;12:1487–93.CrossRefGoogle Scholar
  23. 23.
    Yousef GM, Fracchioli S, Scorilas A, Borgoño CA, Iskander L, Puopolo M, et al. Steroid hormone regulation and prognostic value of the human kallikrein gene 14 in ovarian cancer. Am J Clin Pathol. 2003;119:346–55.CrossRefGoogle Scholar
  24. 24.
    Hashem NN, Mara TW, Mohamed M, Zhang I, Fung K, Kwan KF, et al. Human kallikrein 14 (KLK14) expression in salivary gland tumors. Int J Biol Mark. 2010;25:32–7.CrossRefGoogle Scholar
  25. 25.
    Darling MR, Jackson-Boeters L, Daley TD, Diamandis EP. Human kallikrein 13 expression in salivary gland tumors. Int J Biol Mark. 2006;21(2):106–10.CrossRefGoogle Scholar
  26. 26.
    Darling MR, Tsai S, Jackson-Boeters L, Daley TD, Diamandis EP. Human kallikrein 3 (prostate-specific antigen) and human kallikrein 5 expression in salivary gland tumors. Int J Biol Mark. 2006;21(4):201–5.Google Scholar
  27. 27.
    Darling MR, Hashem NN, Zhang I, Mohamed M, Fung K, Kwan K, et al. Kallikrein-related peptidase 10 expression in salivary gland tissues and tumours. Int J Biol Mark. 2012;27(4):e381–8.CrossRefGoogle Scholar
  28. 28.
    Godfrey TE, Kim SH, Chavira M, Ruff DW, Warren RS, Gray JW, et al. Quantitative mRNA expression analysis from formalin-fixed, paraffin-embedded tissues using 5′ nuclease quantitative reverse transcription-polymerase chain reaction. J Mol Diagn. 2000;2:84–91.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.Interface Oral SurgeryLondonCanada
  2. 2.Pathology and Laboratory Medicine, Schulich School of Medicine and DentistryUniversity of Western OntarioLondonCanada

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