Endocrine Pathology

, Volume 27, Issue 1, pp 1–11 | Cite as

Oncogene-Induced Senescence in Pituitary Adenomas—an Immunohistochemical Study

  • Emilija Manojlovic-Gacic
  • Milica Skender-Gazibara
  • Vera Popovic
  • Ivan Soldatovic
  • Novica Boricic
  • Savo Raicevic
  • Sandra Pekic
  • Mirjana Doknic
  • Dragana Miljic
  • Irina Alafuzoff
  • Fredrik Pontén
  • Olivera Casar-Borota


Oncogene-induced senescence (OIS) serves as an initial barrier to cancer development, being proposed as a possible explanation for the usually benign behavior of the pituitary adenomas. We aimed to explore the immunohistochemical expression of the OIS markers, senescence-associated lysosomal β-galactosidase (SA-β-GAL), p16, and p21 in different types of 345 pituitary adenomas and compared it with the expression in the normal pituitary and in the specimens from the repeated surgeries. SA-β-GAL was overexpressed in the pituitary adenomas, compared to the normal pituitaries. Growth hormone (GH) producing adenomas showed the strongest SA-β-GAL, with densely granulated (DG)-GH adenomas more reactive than the sparsely granulated (SG). Nuclear p21 was decreased in the adenomas, except for the SG-GH adenomas that had higher p21 than the normal pituitaries and the other adenomas. p16 was significantly lower in the adenomas, without type-related differences. SA-β-GAL was slightly lower and p16 slightly higher in the recurrences. Our findings indicate alterations of the senescence program in the different types of pituitary adenomas. Activation of senescence in the pituitary adenomas presents one possible explanation for their usually benign behavior, at least in the GH adenomas that show a synchronous increase of two OIS markers. However, subdivision into GH adenoma subtypes reveals differences that reflect complex regulatory mechanisms influenced by the interplay between the granularity pattern and the hormonal factors, with possible impact on the different clinical behavior of the SG- and DG-GH adenoma subtypes. p16 seems to have a more prominent role in the pituitary tumorigenesis than in the senescence. Recurrent growth in a subset of the pituitary adenomas is not associated with consistent changes in the senescence pattern.


Pituitary adenoma Oncogene-induced senescence GH adenomas granulation pattern Immunohistochemistry 



This work was supported by the Ministry of Education and Science of Republic of Serbia, Grant No 175033. Part of this study performed within the Human Protein Atlas was supported by funding from the Knut and Alice Wallenberg Foundation. We thank Dijana Cerjan for assistance with the immunohistochemical analysis of beta-galactosidase.

Compliance with Ethical Standards

Conflict of Interest

Authors declare that they have no conflict of interest.


  1. 1.
    Xu Y, Li N, Xiang R, Sun P (2014) Emerging roles of the p38 MAPK and PI3K/AKT/mTOR pathways in oncogene-induced senescence. Trends Biochem Sci 39:268–276. doi: 10.1016/j.tibs.2014.04.004 PubMedCentralCrossRefPubMedGoogle Scholar
  2. 2.
    Collado M, Serrano M (2006) The power and the promise of oncogene-induced senescence markers. Nat Rev Cancer 6:472–476. doi: 10.1038/nrc1884 CrossRefPubMedGoogle Scholar
  3. 3.
    Alexandraki KI, Munayem Khan M, Chahal HS, Dalantaeva NS, Trivellin G, Berney DM, Caron P, Popovic V, Pfeifer M, Jordan S, Korbonits M, Grossman AB (2012) Oncogene-induced senescence in pituitary adenomas and carcinomas. Hormones (Athens, Greece) 11:297–307Google Scholar
  4. 4.
    Zhang H (2007) Molecular signaling and genetic pathways of senescence: Its role in tumorigenesis and aging. Journal of Cellular Physiology 210:567–574. doi: 10.1002/jcp.20919 CrossRefPubMedGoogle Scholar
  5. 5.
    Serrano M, Lin AW, McCurrach ME, Beach D, Lowe SW (1997) Oncogenic ras provokes premature cell senescence associated with accumulation of p53 and p16INK4a. Cell 88:593–602CrossRefPubMedGoogle Scholar
  6. 6.
    Quereda V, Martinalbo J, Dubus P, Carnero A, Malumbres M (2007) Genetic cooperation between p21Cip1 and INK4 inhibitors in cellular senescence and tumor suppression. Oncogene 26:7665–7674. doi: 10.1038/sj.onc.1210578 CrossRefPubMedGoogle Scholar
  7. 7.
    Bringold F, Serrano M (2000) Tumor suppressors and oncogenes in cellular senescence. Exp Gerontol 35:317–329CrossRefPubMedGoogle Scholar
  8. 8.
    Campisi J (2005) Senescent cells, tumor suppression, and organismal aging: good citizens, bad neighbors. Cell 120:513–522. doi: 10.1016/j.cell.2005.02.003 CrossRefPubMedGoogle Scholar
  9. 9.
    Campisi J (2013) Aging, cellular senescence, and cancer. Annu Rev Physiol 75:685–705. doi: 10.1146/annurev-physiol-030212-183653 PubMedCentralCrossRefPubMedGoogle Scholar
  10. 10.
    Vousden KH, Lane DP (2007) p53 in health and disease. Nature Reviews Molecular Cell Biology 8:275–283. doi: 10.1038/nrm2147 CrossRefPubMedGoogle Scholar
  11. 11.
    Rodier F, Campisi J, Bhaumik D (2007) Two faces of p53: aging and tumor suppression. Nucleic Acids Res 35:7475–7484. doi: 10.1093/nar/gkm744 PubMedCentralCrossRefPubMedGoogle Scholar
  12. 12.
    Cazzalini O, Scovassi AI, Savio M, Stivala LA, Prosperi E (2010) Multiple roles of the cell cycle inhibitor p21(CDKN1A) in the DNA damage response. Mutat Res 704:12–20. doi: 10.1016/j.mrrev.2010.01.009 CrossRefPubMedGoogle Scholar
  13. 13.
    Rayess H, Wang MB, Srivatsan ES (2012) Cellular senescence and tumor suppressor gene p16. International journal of cancer Journal international du cancer 130 (8):1715–1725. doi: 10.1002/ijc.27316 PubMedCentralCrossRefPubMedGoogle Scholar
  14. 14.
    Bernardes de Jesus B, Blasco MA (2012) Assessing cell and organ senescence biomarkers. Circ Res 111:97–109. doi: 10.1161/circresaha.111.247866 CrossRefPubMedGoogle Scholar
  15. 15.
    Lee BY, Han JA, Im JS, Morrone A, Johung K, Goodwin EC, Kleijer WJ, DiMaio D, Hwang ES (2006) Senescence-associated beta-galactosidase is lysosomal beta-galactosidase. Aging Cell 5:187–195. doi: 10.1111/j.1474-9726.2006.00199.x CrossRefPubMedGoogle Scholar
  16. 16.
    Kurz DJ, Decary S, Hong Y, Erusalimsky JD (2000) Senescence-associated (beta)-galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells. J Cell Sci 113:3613–3622PubMedGoogle Scholar
  17. 17.
    Yang NC, Hu ML (2005) The limitations and validities of senescence associated-beta-galactosidase activity as an aging marker for human foreskin fibroblast Hs68 cells. Exp Gerontol 40:813–819. doi: 10.1016/j.exger.2005.07.011 CrossRefPubMedGoogle Scholar
  18. 18.
    Gary RK, Kindell SM (2005) Quantitative assay of senescence-associated beta-galactosidase activity in mammalian cell extracts. Anal Biochem 343:329–334. doi: 10.1016/j.ab.2005.06.003 CrossRefPubMedGoogle Scholar
  19. 19.
    Kuilman T, Michaloglou C, Mooi WJ, Peeper DS (2010) The essence of senescence. Genes Dev 24:2463–2479. doi: 10.1101/gad.1971610 PubMedCentralCrossRefPubMedGoogle Scholar
  20. 20.
    Debacq-Chainiaux F, Erusalimsky JD, Campisi J, Toussaint O (2009) Protocols to detect senescence-associated beta-galactosidase (SA-betagal) activity, a biomarker of senescent cells in culture and in vivo. Nat Protoc 4:1798–1806. doi: 10.1038/nprot.2009.191 CrossRefPubMedGoogle Scholar
  21. 21.
    Ezzat S, Asa SL, Couldwell WT, Barr CE, Dodge WE, Vance ML, McCutcheon IE (2004) The prevalence of pituitary adenomas: a systematic review. Cancer 101:613–619. doi: 10.1002/cncr.20412 CrossRefPubMedGoogle Scholar
  22. 22.
    Pernicone PJ, Scheithauer BW, Sebo TJ, Kovacs KT, Horvath E, Young WF, Jr., Lloyd RV, Davis DH, Guthrie BL, Schoene WC (1997) Pituitary carcinoma: a clinicopathologic study of 15 cases. Cancer 79:804–812CrossRefPubMedGoogle Scholar
  23. 23.
    Chesnokova V, Zonis S, Rubinek T, Yu R, Ben-Shlomo A, Kovacs K, Wawrowsky K, Melmed S (2007) Senescence mediates pituitary hypoplasia and restrains pituitary tumor growth. Cancer Research 67:10564–10572. doi: 10.1158/0008-5472.can-07-0974 PubMedCentralCrossRefPubMedGoogle Scholar
  24. 24.
    Chesnokova V, Melmed S (2010) Pituitary senescence: the evolving role of Pttg. Mol Cell Endocrinol 326:55–59. doi: 10.1016/j.mce.2010.02.012 PubMedCentralCrossRefPubMedGoogle Scholar
  25. 25.
    Sabatino ME, Petiti JP, Sosa Ldel V, Perez PA, Gutierrez S, Leimgruber C, Latini A, Torres AI, De Paul AL (2015) Evidence of cellular senescence during the development of estrogen-induced pituitary tumors. Endocr Relat Cancer 22:299–317. doi: 10.1530/erc-14-0333 CrossRefPubMedGoogle Scholar
  26. 26.
    Chesnokova V, Zhou C, Ben-Shlomo A, Zonis S, Tani Y, Ren SG, Melmed S (2013) Growth hormone is a cellular senescence target in pituitary and nonpituitary cells. Proceedings of the National Academy of Sciences of the United States of America 110:E3331-3339. doi: 10.1073/pnas.1310589110 PubMedCentralCrossRefPubMedGoogle Scholar
  27. 27.
    Chesnokova V, Zonis S, Kovacs K, Ben-Shlomo A, Wawrowsky K, Bannykh S, Melmed S (2008) p21(Cip1) restrains pituitary tumor growth. Proceedings of the National Academy of Sciences of the United States of America 105:17498–17503. doi: 10.1073/pnas.0804810105 PubMedCentralCrossRefPubMedGoogle Scholar
  28. 28.
    DeLellis RA (2004) Pathology and genetics of tumours of endocrine organs. World Health Organization classification of tumours, vol 8. IARC Press, LyonGoogle Scholar
  29. 29.
    Manojlovic Gacic E, Skender-Gazibara M, Soldatovic I, Dundjerovic D, Boricic N, Raicevic S, Popovic V (2015) Immunohistochemical expression of p16 and p21 in pituitary tissue adjacent to pituitary adenoma versus pituitary tissue obtained at autopsy: is there a difference? Endocrine Pathology 26:104–110. doi: 10.1007/s12022-015-9358-7 CrossRefPubMedGoogle Scholar
  30. 30.
    Kononen J, Bubendorf L, Kallioniemi A, Barlund M, Schraml P, Leighton S, Torhorst J, Mihatsch MJ, Sauter G, Kallioniemi OP (1998) Tissue microarrays for high-throughput molecular profiling of tumor specimens. Nature Medicine 4:844–847CrossRefPubMedGoogle Scholar
  31. 31.
    Uhlen M, Fagerberg L, Hallstrom BM, Lindskog C, Oksvold P, Mardinoglu A, Sivertsson A, Kampf C, Sjostedt E, Asplund A, Olsson I, Edlund K, Lundberg E, Navani S, Szigyarto CA, Odeberg J, Djureinovic D, Takanen JO, Hober S, Alm T, Edqvist PH, Berling H, Tegel H, Mulder J, Rockberg J, Nilsson P, Schwenk JM, Hamsten M, von Feilitzen K, Forsberg M, Persson L, Johansson F, Zwahlen M, von Heijne G, Nielsen J, Ponten F (2015) Proteomics. Tissue-based map of the human proteome. Science (New York, NY) 347:1260419. doi: 10.1126/science.1260419
  32. 32.
    Kampf C, Olsson I, Ryberg U, Sjostedt E, Ponten F (2012) Production of tissue microarrays, immunohistochemistry staining and digitalization within the human protein atlas. Journal of Visualized Experiments : JoVE. doi: 10.3791/3620 PubMedCentralPubMedGoogle Scholar
  33. 33.
    Prasad K, Prabhu GK (2012) Image analysis tools for evaluation of microscopic views of immunohistochemically stained specimen in medical research-a review. J Med Syst 36:2621–2631. doi: 10.1007/s10916-011-9737-7 CrossRefPubMedGoogle Scholar
  34. 34.
    Obari A, Sano T, Ohyama K, Kudo E, Qian ZR, Yoneda A, Rayhan N, Mustafizur Rahman M, Yamada S (2008) Clinicopathological features of growth hormone-producing pituitary adenomas: difference among various types defined by cytokeratin distribution pattern including a transitional form. Endocrine Pathology 19:82–91. doi: 10.1007/s12022-008-9029-z CrossRefPubMedGoogle Scholar
  35. 35.
    Bakhtiar Y, Hirano H, Arita K, Yunoue S, Fujio S, Tominaga A, Sakoguchi T, Sugiyama K, Kurisu K, Yasufuku-Takano J, Takano K (2010) Relationship between cytokeratin staining patterns and clinico-pathological features in somatotropinomae. European journal of endocrinology / European Federation of Endocrine Societies 163:531–539. doi: 10.1530/eje-10-0586 CrossRefPubMedGoogle Scholar
  36. 36.
    Bhayana S, Booth GL, Asa SL, Kovacs K, Ezzat S (2005) The implication of somatotroph adenoma phenotype to somatostatin analog responsiveness in acromegaly. J Clin Endocrinol Metab 90:6290–6295. doi: 10.1210/jc.2005-0998 CrossRefPubMedGoogle Scholar
  37. 37.
    Fougner SL, Casar-Borota O, Heck A, Berg JP, Bollerslev J (2012) Adenoma granulation pattern correlates with clinical variables and effect of somatostatin analogue treatment in a large series of patients with acromegaly. Clin Endocrinol (Oxf) 76 (1):96–102. doi: 10.1111/j.1365-2265.2011.04163.x CrossRefGoogle Scholar
  38. 38.
    Cooper O (2015) Silent corticotroph adenomas. Pituitary 18:225–231. doi: 10.1007/s11102-014-0624-3 CrossRefPubMedGoogle Scholar
  39. 39.
    Saeger W, Ludecke DK, Buchfelder M, Fahlbusch R, Quabbe HJ, Petersenn S (2007) Pathohistological classification of pituitary tumors: 10 years of experience with the German Pituitary Tumor Registry. European journal of endocrinology / European Federation of Endocrine Societies 156:203–216. doi: 10.1530/eje.1.02326 CrossRefPubMedGoogle Scholar
  40. 40.
    Aflorei ED, Korbonits M (2014) Epidemiology and etiopathogenesis of pituitary adenomas. J Neurooncol 117:379–394. doi: 10.1007/s11060-013-1354-5 CrossRefPubMedGoogle Scholar
  41. 41.
    Warfel NA, El-Deiry WS (2013) p21WAF1 and tumourigenesis: 20 years after. Current Opinion in Oncology 25:52–58. doi: 10.1097/CCO.0b013e32835b639e CrossRefPubMedGoogle Scholar
  42. 42.
    Bianchi-Smiraglia A, Nikiforov MA (2012) Controversial aspects of oncogene-induced senescence. Cell Cycle (Georgetown, Tex) 11:4147–4151. doi: 10.4161/cc.22589
  43. 43.
    Laberge RM, Awad P, Campisi J, Desprez PY (2012) Epithelial-mesenchymal transition induced by senescent fibroblasts. Cancer Microenviron 5:39–44. doi: 10.1007/s12307-011-0069-4 PubMedCentralCrossRefPubMedGoogle Scholar
  44. 44.
    Lekva T, Berg JP, Heck A, Lyngvi Fougner S, Olstad OK, Ringstad G, Bollerslev J, Ueland T (2013) Attenuated RORC expression in the presence of EMT progression in somatotroph adenomas following treatment with somatostatin analogs is associated with poor clinical recovery. PLoS One 8:e66927. doi: 10.1371/journal.pone.0066927 PubMedCentralCrossRefPubMedGoogle Scholar
  45. 45.
    Ogino A, Yoshino A, Katayama Y, Watanabe T, Ota T, Komine C, Yokoyama T, Fukushima T (2005) The p15(INK4b)/p16(INK4a)/RB1 pathway is frequently deregulated in human pituitary adenomas. Journal of Neuropathology and Experimental Neurology 64:398–403CrossRefPubMedGoogle Scholar
  46. 46.
    Seemann N, Kuhn D, Wrocklage C, Keyvani K, Hackl W, Buchfelder M, Fahlbusch R, Paulus W (2001) CDKN2A/p16 inactivation is related to pituitary adenoma type and size. The Journal of Pathology 193:491–497. doi: 10.1002/path.833 CrossRefPubMedGoogle Scholar
  47. 47.
    Yoshino A, Katayama Y, Ogino A, Watanabe T, Yachi K, Ohta T, Komine C, Yokoyama T, Fukushima T (2007) Promoter hypermethylation profile of cell cycle regulator genes in pituitary adenomas. J Neurooncol 83:153–162. doi: 10.1007/s11060-006-9316-9 CrossRefPubMedGoogle Scholar
  48. 48.
    Farrell WE (2014) Epigenetics of pituitary tumours: an update. Curr Opin Endocrinol Diabetes Obes 21:299–305. doi: 10.1097/med.0000000000000078 CrossRefPubMedGoogle Scholar
  49. 49.
    Ruebel KH, Jin L, Zhang S, Scheithauer BW, Lloyd RV (2001) Inactivation of the p16 gene in human pituitary nonfunctioning tumors by hypermethylation is more common in null cell adenomas. Endocrine Pathology 12: 281–9CrossRefPubMedGoogle Scholar
  50. 50.
    Rickert CH, Dockhorn-Dworniczak B, Busch G, Moskopp D, Albert FK, Rama B, Paulus W (2001) Increased chromosomal imbalances in recurrent pituitary adenomas. Acta Neuropathologica 102:615–620CrossRefPubMedGoogle Scholar
  51. 51.
    Turner HE, Nagy Z, Sullivan N, Esiri MM, Wass JA (2000) Expression analysis of cyclins in pituitary adenomas and the normal pituitary gland. Clin Endocrinol (Oxf) 53:337–344CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2015

Authors and Affiliations

  • Emilija Manojlovic-Gacic
    • 1
  • Milica Skender-Gazibara
    • 1
  • Vera Popovic
    • 2
    • 3
  • Ivan Soldatovic
    • 3
  • Novica Boricic
    • 1
  • Savo Raicevic
    • 4
  • Sandra Pekic
    • 2
    • 3
  • Mirjana Doknic
    • 2
    • 3
  • Dragana Miljic
    • 2
    • 3
  • Irina Alafuzoff
    • 5
    • 6
  • Fredrik Pontén
    • 7
  • Olivera Casar-Borota
    • 5
    • 6
  1. 1.Institute of Pathology, Medical FacultyUniversity of BelgradeBelgradeSerbia
  2. 2.Clinic of Endocrinology, Diabetes and Metabolic Diseases, Clinical Center of SerbiaBelgradeSerbia
  3. 3.Medical FacultyUniversity of BelgradeBelgradeSerbia
  4. 4.Neurosurgery Clinic, Clinical Center of SerbiaBelgradeSerbia
  5. 5.Department of Immunology, Genetics and PathologyUppsala UniversityUppsalaSweden
  6. 6.Department of Clinical Pathology and CytologyUppsala University HospitalUppsalaSweden
  7. 7.Department of Immunology, Genetics and Pathology, Science for Life Laboratory Uppsala UniversityUppsalaSweden

Personalised recommendations