Tumor Biology

, Volume 37, Issue 3, pp 2909–2924 | Cite as

The roles of parafibromin expression in ovarian epithelial carcinomas: a marker for differentiation and prognosis and a target for gene therapy

  • Dao-fu Shen
  • Xin Liu
  • Xue-feng Yang
  • Lei Fang
  • Yang Gao
  • Shuang Zhao
  • Ji-cheng Wu
  • Shuai Shi
  • Jun-jun Li
  • Xiang-xuan Zhao
  • Wen-feng Gou
  • Hua-chuan Zheng
Original Article


Parafibromin is a protein encoded by hyperparathyroidism 2 (HRPT2) and its downregulated expression is involved in the pathogenesis of parathyroid, breast, gastric, colorectal, lung, head and neck cancers. We aimed to investigate the roles of parafibromin expression in tumorigenesis, progression, or prognostic evaluation of ovarian cancers. HRPT2-expressing plasmid was transfected into ovarian cancer cells with the phenotypes and related molecules examined. The messenger RNA (mRNA) and protein expression of parafibromin were also examined in ovarian normal tissue, benign and borderline tumors and cancers by reverse transcription-polymerase chain reaction (RT-PCR), Western blot, or immunohistochemistry respectively. It was found that parafibromin overexpression caused a lower growth, migration and invasion, higher sensitivity to cisplatin and apoptosis than the mock and control (P < 0.05). The transfectants showed the hypoexpression of phosphoinositide 3-kinase (PI3K), Akt, p70 ribosomal protein S6 kinase (p70s6k), Wnt5a, B cell lymphoma-extra large (Bcl-xL), survivin, vascular endothelial growth factor (VEGF) and matrix metallopeptidase 9 (MMP-9) than the mock and control at both mRNA and protein levels (P < 0.05). According to real-time PCR, parafibromin mRNA level was lower in ovarian benign tumors and cancers than normal ovary (P < 0.05), while parafibromin was strongly expressed in metastatic cancers in omentum than primary cancers by Western blot. Immunohistochemically, parafibromin expression was stronger in primary cancers than that in ovarian normal tissue (P < 0.05) but weaker than the metastatic cancers (P < 0.05) with a positive correlation with dedifferentiation, ki-67 expression and the lower cumulative survival rate (P < 0.05). These findings indicate that parafibromin downregulation might promote the pathogenesis, dedifferentiation and metastasis of ovarian cancers possibly by suppressing aggressive phenotypes, such as proliferation, cell cycle, apoptosis, migration and invasion.


Ovarian cancer Parafibromin Pathogenesis Progression Gene therapy 



This study was supported by President Fund of Liaoning Medical University (XZJJ20140201; XZJJ20140203), a project supported by Scientific Research Fund of Liaoning Provincial Education Department (LJQ2014093), a Key Scientific and Technological Project of Liaoning Province (2015408001), and National Natural Scientific Foundation of China (81172371; 81472544).

Conflicts of interest


Supplementary material

13277_2015_4103_MOESM1_ESM.docx (16 kb)
Supplementary Table 1 (DOCX 16 kb)
13277_2015_4103_MOESM2_ESM.docx (14 kb)
Supplementary Table 2 (DOCX 14 kb)


  1. 1.
    Agarwal SK, Simonds WF, Marx SJ. The parafibromin tumor suppressor protein interacts with actin-binding proteins actinin-2 and actinin-3. Mol Cancer. 2008;7:65.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Bradley KJ, Bowl MR, Williams SE, Ahmad BN, Partridge CJ, Patmanidi AL, et al. Parafibromin is a nuclear protein with a functional monopartite nuclear localization signal. Oncogene. 2007;26:1213–21.CrossRefPubMedGoogle Scholar
  3. 3.
    Bast Jr RC, Hennessy B, Mills GB. The biology of ovarian cancer: new opportunities for translation. Nat Rev Cancer. 2009;9:415–28.CrossRefPubMedPubMedCentralGoogle Scholar
  4. 4.
    Cho KR. Ovarian cancer update: lessons from morphology, molecules, and mice. Arch Pathol Lab Med. 2009;133(11):1775–81.PubMedPubMedCentralGoogle Scholar
  5. 5.
    Cetani F, Banti C, Pardi E, Borsari S, Viacava P, Miccoli P, et al. CDC73 mutational status and loss of parafibromin in the outcome of parathyroid cancer. Endocr Connect. 2013;2:186–95.CrossRefPubMedPubMedCentralGoogle Scholar
  6. 6.
    Gill AJ, Clarkson A, Gimm O, Keil J, Dralle H, Howell VM, et al. Loss of nuclear expression of parafibromin distinguishes parathyroid carcinomas and hyperparathyroidism-jaw tumor (HPT-JT) syndrome-related adenomas from sporadic parathyroid adenomas and hyperplasias. Am J Surg Pathol. 2006;30:1140–9.CrossRefPubMedGoogle Scholar
  7. 7.
    Hahn MA, Dickson KA, Jackson S, Clarkson A, Gill AJ, Marsh DJ, et al. The tumor suppressor CDC73 interacts with the ring finger proteins RNF20 and RNF40 and is required for the maintenance of histone 2B monoubiquitination. Hum Mol Genet. 2012;21:559–68.CrossRefPubMedGoogle Scholar
  8. 8.
    Jo JH, Chung TM, Youn H, Yoo JY. Cytoplasmic parafibromin/hCdc73 targets and destabilizes p53 mRNA to control p53-mediated apoptosis. Nat Commun. 2014;12(5):5433.CrossRefGoogle Scholar
  9. 9.
    Krogan NJ, Dover J, Wood A, Schneider J, Heidt J, Boateng MA, et al. The Paf1 complex is required for histone H3 methylation by COMPASS and Dot1p: linking transcriptional elongation to histone methylation. Mol Cell. 2003;11:721–9.CrossRefPubMedGoogle Scholar
  10. 10.
    Kumada T, Tsuneyama K, Hatta H, Ishizawa S, Takano Y. Improved 1-h rapid immunostaining method using intermittent microwave irradiation: practicability based on 5 years application in Toyama Medical and Pharmaceutical University Hospital. Mod Pathol. 2004;17:1141–9.CrossRefPubMedGoogle Scholar
  11. 11.
    Lin L, Zhang JH, Panicker LM, Simonds WF. The parafibromin tumor suppressor protein inhibits cell proliferation by repression of the c-myc proto-oncogene. Proc Natl Acad Sci U S A. 2008;105:17420–5.CrossRefPubMedPubMedCentralGoogle Scholar
  12. 12.
    Lin L, Czapiga M, Nini L, Zhang JH, Simonds WF. Nuclear localization of the parafibromin tumor suppressor protein implicated in the hyperparathyroidism-jaw tumor syndrome enhances its proapoptotic function. Mol Cancer Res. 2007;5:183–93.CrossRefPubMedGoogle Scholar
  13. 13.
    Mosimann C, Hausmann G, Basler K. Parafibromin/Hyrax activates Wnt/Wg target gene transcription by direct association with beta-catenin/Armadillo. Cell. 2006;125:327–41.CrossRefPubMedGoogle Scholar
  14. 14.
    Newey PJ, Bowl MR, Thakker RV. Parafibromin-functional insights. J Intern Med. 2009;266:84–98.CrossRefPubMedGoogle Scholar
  15. 15.
    Nishita M, Enomoto M, Yamagata K, Minami Y. Cell/tissue-tropic functions of Wnt5a signaling in normal and cancer cells. Trends Cell Biol. 2010;20:346–54.CrossRefPubMedGoogle Scholar
  16. 16.
    Prat J. Ovarian carcinomas: five distinct diseases with different origins, genetic alterations, and clinicopathological features. Virchows Arch. 2012;460:237–49.CrossRefPubMedGoogle Scholar
  17. 17.
    Rather MI, Swamy S, Gopinath KS, Kumar A. Transcriptional repression of tumor suppressor CDC73, encoding an RNA polymerase II interactor, by Wilms tumor 1 protein (WT1) promotes cell proliferation: implication for cancer therapeutics. J Biol Chem. 2014;289:968–76.CrossRefPubMedGoogle Scholar
  18. 18.
    Selvarajan S, Sii LH, Lee A, Yip G, Bay BH, Tan MH, et al. Parafibromin expression in breast cancer: a novel marker for prognostication? J Clin Pathol. 2008;61:64–7.CrossRefPubMedGoogle Scholar
  19. 19.
    Shao JL, Wan XH, Chen Y, Bi C, Chen HM, Zhong Y, et al. H2S protects hippocampal neurons from anoxia-reoxygenation through cAMP-mediated PI3K/Akt/p70S6K cell-survival signaling pathways. J Mol Neurosci. 2011;43:453–60.CrossRefPubMedGoogle Scholar
  20. 20.
    Saiki S, Sasazawa Y, Imamichi Y, Kawajiri S, Fujimaki T, Tanida I, et al. Caffeine induces apoptosis by enhancement of autophagy via PI3K/Akt/mTOR/p70S6K inhibition. Autophagy. 2011;7:176–87.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Truran PP, Johnson SJ, Bliss RD, Lennard TW, Aspinall SR. Parafibromin, galectin-3, PGP9.5, Ki67, and cyclin D1: using an immunohistochemical panel to aid in the diagnosis of parathyroid cancer. World J Surg. 2014;38:2845–54.CrossRefPubMedGoogle Scholar
  22. 22.
    Takahashi A, Tsutsumi R, Kikuchi I, Obuse C, Saito Y, Seidi A, et al. SHP2 tyrosine phosphatase converts parafibromin/Cdc73 from a tumor suppressor to an oncogenic driver. Mol Cell. 2011;43:45–56.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Witteveen JE, Hamdy NA, Dekkers OM, Kievit J, van Wezel T, Teh BT, et al. Downregulation of CASR expression and global loss of parafibromin staining are strong negative determinants of prognosis in parathyroid carcinoma. Mod Pathol. 2011;24:688–97.CrossRefPubMedGoogle Scholar
  24. 24.
    Xia P, Wang W, Xu XY, Wang JP, Takano Y, Zheng HC, et al. Parafibromin expression in lung normal tissue and carcinoma: its comparison with clinicopathological parameters of carcinoma. Histol Histopathol. 2011;26:1039–47.PubMedGoogle Scholar
  25. 25.
    Yart A, Gstaiger M, Wirbelauer C, Pecnik M, Anastasiou D, Hess D, et al. The HRPT2 tumour suppressor gene product parafibromin associates with human PAF1 and RNA polymerase II. Mol Cell Biol. 2007;25:5052–60.CrossRefGoogle Scholar
  26. 26.
    Yang YJ, Han JW, Youn HD, Cho EJ. The tumor suppressor, parafibromin, mediates histone H3 K9 methylation for cyclin D1 repression. Nucleic Acids Res. 2010;38:382–90.CrossRefPubMedGoogle Scholar
  27. 27.
    Zhang Z, Yang XF, Huang KQ, Ren L, Gou WF, Shen DF, Zhao S, Sun HZ, Takano Y, Zheng HC, et al. The clinicopathological significance and biological functions of parafibromin expression in head and neck squamous cell carcinomas. Tum Biol in pressGoogle Scholar
  28. 28.
    Zheng HC, Takahashi H, Li XH, Hara T, Masuda S, Guan YF, et al. Downregulated parafibromin expression is a promising marker for pathogenesis, invasion, metastasis and prognosis of gastric carcinomas. Virchows Arch. 2008;452:147–55.CrossRefPubMedGoogle Scholar
  29. 29.
    Zheng HC, Wei ZL, Xu XY, Nie XC, Yang X, Takahashi H, et al. Parafibromin expression is an independent prognostic factor for colorectal carcinomas. Hum Pathol. 2011;42:1089–102.CrossRefPubMedGoogle Scholar
  30. 30.
    Zheng HC, Nakamura T, Zheng Y, Nakanishi Y, Tabuchi Y, Uchiyama A, et al. SV40 T antigen disrupted the cell metabolism and the balance between proliferation and apoptosis in lens tumors of transgenic mice. J Cancer Res Clin Oncol. 2009;135:1521–32.CrossRefPubMedGoogle Scholar
  31. 31.
    Zhao S, Sun HZ, Zhu ST, Lu H, Niu ZF, Guo WF, et al. Effects of parafibromin expression on the phenotypes and relevant mechanisms in the DLD-1 colon carcinoma cell line. Asian Pac J Cancer Prev. 2013;14:4249–54.CrossRefPubMedGoogle Scholar
  32. 32.
    Zheng H, Tsuneyama K, Cheng C, Takahashi H, Cui Z, Murai Y, et al. An immunohistochemical study of P53 and Ki-67 in gastrointestinal adenoma and adenocarcinoma using tissue microarray. Anticancer Res. 2006;26:2353–60.PubMedGoogle Scholar
  33. 33.
    Zheng H, Takahashi H, Murai Y, Cui Z, Nomoto K, Niwa H, et al. Expressions of MMP-2, MMP-9 and VEGF are closely linked to growth, invasion, metastasis and angiogenesis of gastric carcinoma. Anticancer Res. 2006;26:3579–83.PubMedGoogle Scholar

Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2015

Authors and Affiliations

  • Dao-fu Shen
    • 1
  • Xin Liu
    • 2
  • Xue-feng Yang
    • 1
  • Lei Fang
    • 1
  • Yang Gao
    • 1
  • Shuang Zhao
    • 1
  • Ji-cheng Wu
    • 1
  • Shuai Shi
    • 1
  • Jun-jun Li
    • 1
  • Xiang-xuan Zhao
    • 1
  • Wen-feng Gou
    • 1
  • Hua-chuan Zheng
    • 1
  1. 1.Cancer Center, Key Laboratory of Brain and Spinal Cord Injury of Liaoning Province, and Laboratory Animal CenterThe First Affiliated Hospital of Liaoning Medical UniversityJinzhouChina
  2. 2.Department of Clinical LaboratoryThe First Affiliated Hospital of Liaoning Medical UniversityJinzhouChina

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