Skip to main content

Advertisement

SpringerLink
Log in

Clusterin and Chemotherapy Sensitivity Under Normoxic and Graded Hypoxic Conditions in Colorectal Cancer

  • Original Article
  • Published:
Journal of Gastrointestinal Cancer Aims and scope Submit manuscript

Abstract

Background and Aims

In vitro studies have shown that clusterin modulates treatment sensitivity in a number of human cancers; however, the interaction between clusterin expression and hypoxia in controlling treatment response in CRC has not previously been examined. The aim of this study was to assess the effect of clusterin overexpression in CRC cells on sensitivity to 5-fluorouracil (5-FU), oxaliplatin and FOLFOX treatment under normoxic and graded hypoxic conditions.

Methods

SW480 colon cancer cells were transfected with full length Clusterin cDNA to generate a clusterin overexpressing cell line. Overexpression was confirmed by western blot analysis. The response of parental and clusterin overexpressing cells to 5-FU, oxaliplatin and FOLFOX was examined using a crystal violet-based proliferation assay under normoxic conditions, 3% and 1% hypoxic conditions. The levels of apoptosis and G2/M arrest in FOLFOX-treated cells were assessed by flow cytometry.

Results

Under normoxic conditions, clusterin overexpressing cells were more sensitive to FOLFOX treatment (p = 0.01); under 3% and 1% hypoxic conditions, overexpressing clusterin cells were more sensitive to 5-FU, oxaliplatin and FOLFOX, p values <0.05 for all conditions. Under normoxic conditions, overexpressing clusterin cells showed significantly higher levels of apoptosis when treated with FOLFOX compared to untransfected cells; levels of G2M cells were not significantly different. Under both 3% and 1% hypoxia, the percentage of cells undergoing apoptosis following FOLFOX treatment was significantly higher in overexpressing clusterin cells.

Conclusion

These in vitro findings suggest that tumours expressing high levels of clusterin, particularly if hypoxic in nature, may benefit from treatments such as FOLFOX.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3

Similar content being viewed by others

References

  1. Trougakos IP, Gonos ES. Clusterin/apolipoprotein J in human aging and cancer. Int J Biochem Cell Biol. 2002;34(11):1430–48.

    Article  PubMed  CAS  Google Scholar 

  2. Pucci S, Bonanno E, Pichiorri F, Angeloni C, Spagnoli LG. Modulation of different clusterin isoforms in human colon tumorigenesis. Oncogene. 2004;23(13):2298–304. doi:10.1038/sj.onc.1207404.

    Article  PubMed  CAS  Google Scholar 

  3. Shannan B, Seifert M, Leskov K, Willis J, Boothman D, Tilgen W, et al. Challenge and promise: roles for clusterin in pathogenesis, progression and therapy of cancer. Cell Death Differ. 2006;13(1):12–9. doi:10.1038/sj.cdd.4401779.

    Article  PubMed  CAS  Google Scholar 

  4. Andersen CL, Schepeler T, Thorsen K, Birkenkamp-Demtroder K, Mansilla F, Aaltonen LA, et al. Clusterin expression in normal mucosa and colorectal cancer. Mol Cell Proteomics. 2007;6(6):1039–48. doi:10.1074/mcp.M600261-MCP200.

    Article  PubMed  CAS  Google Scholar 

  5. Lakins J, Bennett SA, Chen JH, Arnold JM, Morrissey C, Wong P, et al. Clusterin biogenesis is altered during apoptosis in the regressing rat ventral prostate. J Biol Chem. 1998;273(43):27887–95.

    Article  PubMed  CAS  Google Scholar 

  6. O’Sullivan J, Whyte L, Drake J, Tenniswood M. Alterations in the post-translational modification and intracellular trafficking of clusterin in MCF-7 cells during apoptosis. Cell Death Differ. 2003;10(8):914–27. doi:10.1038/sj.cdd.4401254.

    Article  PubMed  Google Scholar 

  7. Wilson MR, Easterbrook-Smith SB. Clusterin binds by a multivalent mechanism to the Fc and Fab regions of IgG. Biochim Biophys Acta. 1992;1159(3):319–26.

    Article  PubMed  CAS  Google Scholar 

  8. de Silva HV, Harmony JA, Stuart WD, Gil CM, Robbins J. Apolipoprotein J: structure and tissue distribution. Biochemistry. 1990;29(22):5380–9.

    Article  PubMed  Google Scholar 

  9. Jenne DE, Lowin B, Peitsch MC, Bottcher A, Schmitz G, Tschopp J. Clusterin (complement lysis inhibitor) forms a high density lipoprotein complex with apolipoprotein A-I in human plasma. J Biol Chem. 1991;266(17):11030–6.

    PubMed  CAS  Google Scholar 

  10. Li DQ, Ljungh A. Binding of human clusterin by Staphylococcus epidermidis. FEMS Immunol Med Microbiol. 2001;31(3):197–202.

    Article  PubMed  CAS  Google Scholar 

  11. Matsubara E, Soto C, Governale S, Frangione B, Ghiso J. Apolipoprotein J and Alzheimer’s amyloid beta solubility. Biochem J. 1996;316(Pt 2):671–9.

    PubMed  CAS  Google Scholar 

  12. Kujiraoka T, Hattori H, Miwa Y, Ishihara M, Ueno T, Ishii J, et al. Serum apolipoprotein j in health, coronary heart disease and type 2 diabetes mellitus. J Atheroscler Thromb. 2006;13(6):314–22.

    Article  PubMed  CAS  Google Scholar 

  13. Bajari TM, Strasser V, Nimpf J, Schneider WJ. A model for modulation of leptin activity by association with clusterin. FASEB J. 2003;17(11):1505–7. doi:10.1096/fj.02-1106fje.

    PubMed  CAS  Google Scholar 

  14. Matsuda A, Itoh Y, Koshikawa N, Akizawa T, Yana I, Seiki M. Clusterin, an abundant serum factor, is a possible negative regulator of MT6-MMP/MMP-25 produced by neutrophils. J Biol Chem. 2003;278(38):36350–7. doi:10.1074/jbc.M301509200.

    Article  PubMed  CAS  Google Scholar 

  15. Leskov KS, Klokov DY, Li J, Kinsella TJ, Boothman DA. Synthesis and functional analyses of nuclear clusterin, a cell death protein. J Biol Chem. 2003;278(13):11590–600. doi:10.1074/jbc.M209233200.

    Article  PubMed  CAS  Google Scholar 

  16. Miyake H, Nelson C, Rennie PS, Gleave ME. Acquisition of chemoresistant phenotype by overexpression of the antiapoptotic gene testosterone-repressed prostate message-2 in prostate cancer xenograft models. Cancer Res. 2000;60(9):2547–54.

    PubMed  CAS  Google Scholar 

  17. Cervellera M, Raschella G, Santilli G, Tanno B, Ventura A, Mancini C, et al. Direct transactivation of the anti-apoptotic gene apolipoprotein J (clusterin) by B-MYB. J Biol Chem. 2000;275(28):21055–60. doi:10.1074/jbc.M002055200.

    Article  PubMed  CAS  Google Scholar 

  18. Zellweger T, Chi K, Miyake H, Adomat H, Kiyama S, Skov K, et al. Enhanced radiation sensitivity in prostate cancer by inhibition of the cell survival protein clusterin. Clin Cancer Res. 2002;8(10):3276–84.

    PubMed  CAS  Google Scholar 

  19. Muramaki M, So A, Hayashi N, Sowery R, Miyake H, Fujisawa M, et al. Chemosensitization of gemcitabine-resistant human bladder cancer cell line both in vitro and in vivo using antisense oligonucleotide targeting the anti-apoptotic gene, clusterin. BJU Int. 2009;103(3):384–90. doi:10.1111/j.1464-410X.2008.08098.x.

    Article  PubMed  CAS  Google Scholar 

  20. Yamanaka K, Gleave M, Muramaki M, Hara I, Miyake H. Enhanced radiosensitivity by inhibition of the anti-apoptotic gene clusterin using antisense oligodeoxynucleotide in a human bladder cancer model. Oncol Rep. 2005;13(5):885–90.

    PubMed  CAS  Google Scholar 

  21. Toffanin S, Daidone MG, Miodini P, De Cecco L, Gandellini P, Cappelletti V. Clusterin: a potential target for improving response to antiestrogens. Int J Oncol. 2008;33(4):791–8.

    PubMed  CAS  Google Scholar 

  22. Gleave M, Miyake H. Use of antisense oligonucleotides targeting the cytoprotective gene, clusterin, to enhance androgen- and chemo-sensitivity in prostate cancer. World J Urol. 2005;23(1):38–46. doi:10.1007/s00345-004-0474-0.

    Article  PubMed  CAS  Google Scholar 

  23. Vaupel P, Thews O, Hoeckel M. Treatment resistance of solid tumors: role of hypoxia and anemia. Med Oncol. 2001;18(4):243–59. doi:10.1385/MO:18:4:243.

    Article  PubMed  CAS  Google Scholar 

  24. Vaupel P, Mayer A, Hockel M. Tumor hypoxia and malignant progression. Methods Enzymol. 2004;381:335–54. doi:10.1016/S0076-6879(04)81023-1.

    Article  PubMed  CAS  Google Scholar 

  25. Trougakos IP, So A, Jansen B, Gleave ME, Gonos ES. Silencing expression of the clusterin/apolipoprotein j gene in human cancer cells using small interfering RNA induces spontaneous apoptosis, reduced growth ability, and cell sensitization to genotoxic and oxidative stress. Cancer Res. 2004;64(5):1834–42.

    Article  PubMed  CAS  Google Scholar 

  26. Bettuzzi S, Scorcioni F, Astancolle S, Davalli P, Scaltriti M, Corti A. Clusterin (SGP-2) transient overexpression decreases proliferation rate of SV40-immortalized human prostate epithelial cells by slowing down cell cycle progression. Oncogene. 2002;21(27):4328–34. doi:10.1038/sj.onc.1205594.

    Article  PubMed  CAS  Google Scholar 

  27. Yang CR, Leskov K, Hosley-Eberlein K, Criswell T, Pink JJ, Kinsella TJ, et al. Nuclear clusterin/XIP8, an X-ray-induced Ku70-binding protein that signals cell death. Proc Natl Acad Sci USA. 2000;97(11):5907–12.

    Article  PubMed  CAS  Google Scholar 

  28. Yotnda P, Wu D, Swanson AM. Hypoxic tumors and their effect on immune cells and cancer therapy. Methods Mol Biol. 2010;651:1–29. doi:10.1007/978-1-60761-786-0_1.

    Article  PubMed  CAS  Google Scholar 

  29. Giatromanolaki A, Harris AL. Tumour hypoxia, hypoxia signaling pathways and hypoxia inducible factor expression in human cancer. Anticancer Res. 2001;21(6B):4317–24.

    PubMed  CAS  Google Scholar 

  30. Voss MJ, Niggemann B, Zanker KS, Entschladen F. Tumour reactions to hypoxia. Curr Mol Med. 2010;10(4):381–6.

    Article  PubMed  CAS  Google Scholar 

  31. Brahimi-Horn MC, Chiche J, Pouyssegur J. Hypoxia and cancer. J Mol Med. 2007;85(12):1301–7. doi:10.1007/s00109-007-0281-3.

    Article  PubMed  Google Scholar 

  32. Vaupel P. The role of hypoxia-induced factors in tumor progression. Oncologist. 2004;9 Suppl 5:10–7. doi:10.1634/theoncologist.9-90005-10.

    Article  PubMed  CAS  Google Scholar 

  33. Liao D, Johnson RS. Hypoxia: a key regulator of angiogenesis in cancer. Cancer Metastasis Rev. 2007;26(2):281–90. doi:10.1007/s10555-007-9066-y.

    Article  PubMed  CAS  Google Scholar 

  34. Kim JW, Gao P, Dang CV. Effects of hypoxia on tumor metabolism. Cancer Metastasis Rev. 2007;26(2):291–8. doi:10.1007/s10555-007-9060-4.

    Article  PubMed  Google Scholar 

  35. Cerniglia GJ, Pore N, Tsai JH, Schultz S, Mick R, Choe R, et al. Epidermal growth factor receptor inhibition modulates the microenvironment by vascular normalization to improve chemotherapy and radiotherapy efficacy. PLoS ONE. 2009;4(8):e6539. doi:10.1371/journal.pone.0006539.

    Article  PubMed  Google Scholar 

  36. Tamm M, Bihl M, Eickelberg O, Stulz P, Perruchoud AP, Roth M. Hypoxia-induced interleukin-6 and interleukin-8 production is mediated by platelet-activating factor and platelet-derived growth factor in primary human lung cells. Am J Respir Cell Mol Biol. 1998;19(4):653–61.

    PubMed  CAS  Google Scholar 

  37. Baggiolini M, Clark-Lewis I. Interleukin-8, a chemotactic and inflammatory cytokine. FEBS Lett. 1992;307(1):97–101.

    Article  PubMed  CAS  Google Scholar 

  38. Ning Y, Manegold PC, Hong YK, Zhang W, Pohl A, Lurje G, et al. Interleukin-8 is associated with proliferation, migration, angiogenesis and chemosensitivity in vitro and in vivo in colon cancer cell line models. Int J Cancer. 2011;128(9):2038–49. doi:10.1002/ijc.25562.

    Article  PubMed  CAS  Google Scholar 

  39. Wang Y, Guo XQ, Niu XL, Wu J, Zhu YQ, Mao LQ. Relationship of IL-6 and IL-8 secretion in epithelial ovarian cancer cell lines with their sensitivity to tamoxifen as well as MAPK, Akt and estrogen receptor phosphorylation. Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2010;26(1):21–4.

    PubMed  Google Scholar 

  40. Wilson C, Wilson T, Johnston PG, Longley DB, Waugh DJ. Interleukin-8 signaling attenuates TRAIL- and chemotherapy-induced apoptosis through transcriptional regulation of c-FLIP in prostate cancer cells. Mol Cancer Ther. 2008;7(9):2649–61. doi:10.1158/1535-7163.MCT-08-0148.

    Article  PubMed  CAS  Google Scholar 

  41. Miyake H, Yamanaka K, Muramaki M, Kurahashi T, Gleave M, Hara I. Enhanced expression of the secreted form of clusterin following neoadjuvant hormonal therapy as a prognostic predictor in patients undergoing radical prostatectomy for prostate cancer. Oncol Rep. 2005;14(5):1371–5.

    PubMed  CAS  Google Scholar 

  42. Kurahashi T, Muramaki M, Yamanaka K, Hara I, Miyake H. Expression of the secreted form of clusterin protein in renal cell carcinoma as a predictor of disease extension. BJU Int. 2005;96(6):895–9. doi:10.1111/j.1464-410X.2005.05733.x.

    Article  PubMed  CAS  Google Scholar 

  43. Xie D, Lau SH, Sham JS, Wu QL, Fang Y, Liang LZ, et al. Up-regulated expression of cytoplasmic clusterin in human ovarian carcinoma. Cancer. 2005;103(2):277–83. doi:10.1002/cncr.20765.

    Article  PubMed  Google Scholar 

  44. Watari H, Ohta Y, Hassan MK, Xiong Y, Tanaka S, Sakuragi N. Clusterin expression predicts survival of invasive cervical cancer patients treated with radical hysterectomy and systematic lymphadenectomy. Gynecol Oncol. 2008;108(3):527–32. doi:10.1016/j.ygyno.2007.11.026.

    Article  PubMed  CAS  Google Scholar 

  45. Kang YK, Hong SW, Lee H, Kim WH. Overexpression of clusterin in human hepatocellular carcinoma. Hum Pathol. 2004;35(11):1340–6. doi:10.1016/j.humpath.2004.07.021.

    Article  PubMed  CAS  Google Scholar 

  46. Xie MJ, Motoo Y, Su SB, Mouri H, Ohtsubo K, Matsubara F, et al. Expression of clusterin in human pancreatic cancer. Pancreas. 2002;25(3):234–8.

    Article  PubMed  Google Scholar 

  47. Albert JM, Gonzalez A, Massion PP, Chen H, Olson SJ, Shyr Y, et al. Cytoplasmic clusterin expression is associated with longer survival in patients with resected non small cell lung cancer. Cancer Epidemiol Biomarkers Prev. 2007;16(9):1845–51. doi:10.1158/1055-9965.EPI-07-0146.

    Article  PubMed  CAS  Google Scholar 

  48. Kevans D, Foley J, Tenniswood M, Sheahan K, Hyland J, O’Donoghue D, et al. High clusterin expression correlates with a poor outcome in stage II colorectal cancers. Cancer Epidemiol Biomarkers Prev. 2009;18(2):393–9. doi:10.1158/1055-9965.EPI-08-0302.

    Article  PubMed  CAS  Google Scholar 

Download references

Disclosures

The authors declare that they have no conflict of interest or financial support to disclose in relation to this article.

Author information

Authors and Affiliations

  1. Centre for Colorectal Disease, St. Vincent’s University Hospital, Elm Park, Dublin 4, Ireland

    David Kevans, Sheeona Gorman, Miriam Tosetto, Kieran Sheahan, Diarmuid O’Donoghue, Hugh Mulcahy & Jacintha O’Sullivan

  2. University College Dublin, Dublin, Ireland

    David Kevans, Sheeona Gorman, Miriam Tosetto, Kieran Sheahan, Diarmuid O’Donoghue, Hugh Mulcahy & Jacintha O’Sullivan

Authors
  1. David Kevans
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Sheeona Gorman
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Miriam Tosetto
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Kieran Sheahan
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Diarmuid O’Donoghue
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Hugh Mulcahy
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Jacintha O’Sullivan
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Jacintha O’Sullivan.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kevans, D., Gorman, S., Tosetto, M. et al. Clusterin and Chemotherapy Sensitivity Under Normoxic and Graded Hypoxic Conditions in Colorectal Cancer. J Gastrointest Canc 43, 305–313 (2012). https://doi.org/10.1007/s12029-011-9277-x

Download citation

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s12029-011-9277-x

Keywords

Search

Navigation