Clinical & Experimental Metastasis

, Volume 32, Issue 5, pp 441–455 | Cite as

Transketolase is upregulated in metastatic peritoneal implants and promotes ovarian cancer cell proliferation

  • Carmela Ricciardelli
  • Noor A. Lokman
  • Sowmya Cheruvu
  • Izza A. Tan
  • Miranda P. Ween
  • Carmen E. Pyragius
  • Andrew Ruszkiewicz
  • Peter Hoffmann
  • Martin K. Oehler
Article

Abstract

Ovarian cancer, the most lethal gynaecological cancer, is characterised by the shedding of epithelial cells from the ovarian surface, followed by metastasis and implantation onto the peritoneal surfaces of abdominal organs. Our proteomic studies investigating the interactions between peritoneal (LP-9) and ovarian cancer (OVCAR-5) cells found transketolase (TKT) to be regulated in the co-culture system. This study characterized TKT expression in advanced stage (III/IV) serous ovarian cancers (n = 125 primary and n = 54 peritoneal metastases), normal ovaries (n = 6) and benign serous cystadenomas (n = 10) by immunohistochemistry. In addition, we also evaluated the function of TKT in ovarian cancer cells in vitro. Nuclear TKT was present in all primary serous ovarian cancer tissues examined (median 82.0 %, range 16.5–100 %) and was significantly increased in peritoneal metastases compared with matching primary cancers (P = 0.01, Wilcoxon Rank test). Kaplan–Meier survival and Cox regression analyses showed that high nuclear TKT positivity in peritoneal metastases (>94 %) was significantly associated with reduced overall survival (P = 0.006) and a 2.8 fold increased risk of ovarian cancer death (95 % CI 1.29–5.90, P = 0.009). Knockdown of TKT by siRNAs significantly reduced SKOV-3 cell proliferation but had no effect on their motility or invasion. Oxythiamine, an inhibitor of TKT activity, significantly inhibited the proliferation of four ovarian cancer cell lines (OV-90, SKOV-3, OVCAR-3 and OVCAR-5) and primary serous ovarian cancer cells isolated from patient ascites. In conclusion, these findings indicate that TKT plays an important role in the proliferation of metastatic ovarian cancer cells and could be used as novel therapeutic target for advanced disease.

Keywords

Ovarian cancer Transketolase Glucose metabolism Pentose phosphate pathway Oxythiamine Invasion Motility Proliferation Metastasis 

Abbreviations

CM

Conditioned media

FBS

Fetal bovine serum

PPP

Phosphate pentose pathway

TKT

Transketolase

TDP

Thiamine diphosphate

Notes

Acknowledgments

We thank Dr. Thomas Hamilton (Fox Chase Cancer Center, Philadelphia, PA) for kindly providing the OVCAR-5 cell line, Mrs Wendy Bonner and Dr Noor Hammodi for their help with the TKT immunostaining. This research has been funded by the Ovarian Cancer Research Foundation (OCRF), Australia, Cancer Council of South Australia and South Australian Health and Medical Research Institute.

Conflict of interest

The authors declare they have no conflict of interest.

Supplementary material

10585_2015_9718_MOESM1_ESM.tif (3.8 mb)
Supplementary material 1 (TIFF 3891 kb). Supplementary Fig. 1: Cytokeratin immunocytochemical staining in primary serous ovarian cancer cells. Pan-cytokeratin immunostaining of primary ovarian cancer cells derived from patient 1 (a) and patient 2 (b) ascites compared with the established serous ovarian cancer cell line OV-90 (c). d Represents OV-90 cells negative control of immunocytochemistry without primary antibody. Magnification bar = 100 µm for all images
10585_2015_9718_MOESM2_ESM.tif (799 kb)
Supplementary material 2 (TIFF 798 kb). Supplementary Fig. 2: Ion-trap mass spectrometry LC–MS/MS analysis of spot 1 (a) and spot 2 (b) identified in the secretome of co-cultured OVCAR-5 and LP-9 cells by 2D gel electrophoresis and silver staining
10585_2015_9718_MOESM3_ESM.docx (16 kb)
Supplementary material 3 (DOCX 16 kb). Supplementary Table 1: Summary of clinical and pathological characteristics of the primary ovarian cancer cells established from patient ascites

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Copyright information

© Springer Science+Business Media Dordrecht 2015

Authors and Affiliations

  • Carmela Ricciardelli
    • 1
  • Noor A. Lokman
    • 1
  • Sowmya Cheruvu
    • 1
  • Izza A. Tan
    • 1
  • Miranda P. Ween
    • 2
  • Carmen E. Pyragius
    • 1
  • Andrew Ruszkiewicz
    • 3
    • 4
  • Peter Hoffmann
    • 5
  • Martin K. Oehler
    • 1
    • 6
  1. 1.Discipline of Obstetrics and Gynaecology, School of Paediatrics and Reproductive Health, Robinson Research InstituteUniversity of AdelaideAdelaideAustralia
  2. 2.Research Centre for Infectious Diseases, School of Molecular BiosciencesUniversity of AdelaideAdelaideAustralia
  3. 3.Centre of Cancer BiologyUniversity of South AustraliaAdelaideAustralia
  4. 4.Department of Anatomical PathologySA PathologyAdelaideAustralia
  5. 5.Adelaide Proteomics Centre, School of Molecular and Biomedical ScienceUniversity of AdelaideAdelaideAustralia
  6. 6.Department of Gynaecological OncologyRoyal Adelaide HospitalAdelaideAustralia

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