Skip to main content

Advertisement

SpringerLink
Log in

Epigenetic control of group V phospholipase A2 expression in human malignant cells

  • Original Article
  • Published:
Tumor Biology

Abstract

Secreted phospholipases A2 (sPLA2) are suggested to play an important role in inflammation and tumorigenesis. Different mechanisms of epigenetic regulation are involved in the control of group IIA, III and X sPLA2s expression in cancer cells, but group V sPLA2 (GV-PLA2) in this respect has not been studied. Here, we demonstrate the role of epigenetic mechanisms in regulation of GV-PLA2 expression in different cell lines originating from leukaemia and solid cancers. In blood leukocytes from leukaemic patients, levels of GV-PLA2 transcripts were significantly lower in comparison to those from healthy individuals. Similarly, in DU-145 and PC-3 prostate and CAL-51 and MCF-7 mammary cancer cell lines, levels of GV-PLA2 transcripts were significantly lower in relation to those found in normal epithelial cells of prostate or mammary. By sequencing and methylation-specific high-resolution melting (MS-HRM) analyses of bisulphite-modified DNA, distinct CpG sites in the GV-PLA2 promoter region were identified that were differentially methylated in cancer cells in comparison to normal epithelial and endothelial cells. Spearman rank order analysis revealed a significant negative correlation between the methylation degree and the cellular expression of GV-PLA2 (r = −0.697; p = 0.01). The effects of demethylating agent (5-aza-2′-deoxycytidine) and histone deacetylase inhibitor (trichostatin A) on GV-PLA2 transcription in the analysed cells confirmed the importance of DNA methylation and histone modification in the regulation of the GV-PLA2 gene expression in leukaemic, prostate and mammary cancer cell lines. The exposure of tumour cells to human recombinant GV-PLA2 resulted in a reduced colony forming activity of MCF-7, HepG2 and PC-3 cells, but not of DU-145 cells suggesting a cell-type-dependent effect of GV-PLA2 on cell growth. In conclusion, our results suggest that epigenetic mechanisms such as DNA methylation and histone modification play an important role in downregulation of GV-PLA2 expression in cancer cells.

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
Fig. 4
Fig. 5

Similar content being viewed by others

Abbreviations

5-aza-dC:

5-Aza-2′-deoxycytidine

CV:

Crystal violet

DMSO:

Dimethyl sulfoxide

FCS:

Foetal calf serum

GV-PLA2 :

Group V secreted phospholipase A2

HCAEC:

Human coronary artery endothelial cells

HUVEC:

Human umbilical vein endothelial cells

MAPK:

Mitogen-activated protein kinase

MS-HRM:

Methylation-specific high-resolution melting

PBS:

Phosphate-buffered saline

PCR:

Polymerase chain reaction

RT-qPCR:

Real-time quantitative PCR

SEM:

Standard error of the mean

sPLA2 :

Secreted phospholipase A2

TSA:

Trichostatin A

References

  1. Dennis EA, Deems RA, Yu L. Extracellular phospholipase A2. Adv Exp Med Biol. 1992;318:35–9.

    Article  CAS  PubMed  Google Scholar 

  2. Murakami M, Lambeau G. Emerging roles of secreted phospholipase A2 enzymes: an update. Biochimie. 2013;95:43–50.

    Article  CAS  PubMed  Google Scholar 

  3. Menschikowski M, Hagelgans A, Siegert G. Secretory phospholipase A2 of group IIA: is it an offensive or a defensive player during atherosclerosis and other inflammatory diseases? Prostaglandins Other Lipid Mediat. 2006;79:1–33.

    Article  CAS  PubMed  Google Scholar 

  4. Chen J, Engle SJ, Seilhamer JJ, Tischfield JA. Cloning and recombinant expression of a novel human low molecular weight Ca2+-dependent phospholipase A2. J Biol Chem. 1994;269:2365–8.

    CAS  PubMed  Google Scholar 

  5. Xue M, Shen K, McKelvey K, Li J, Chan YK, Hatzis V, et al. Endothelial protein C receptor associated invasiveness of rheumatoid synovial fibroblasts is likely driven by group V secretory phospholipase A2. Arthritis Res Ther. 2014;16:R44.

    Article  PubMed  PubMed Central  Google Scholar 

  6. Rosengren B, Peilot H, Umaerus M, Jönsson-Rylander AC, Mattsson-Hultén L, Hallberg C, et al. Secretory phospholipase A2 group V: lesion distribution, activation by arterial proteoglycans and induction in aorta by a western diet. Arterioscler Thromb Vasc Biol. 2006;26:1579–85.

    Article  CAS  PubMed  Google Scholar 

  7. Sonoki K, Iwase M, Ohdo S, Ieiri I, Matsuyama N, Takata Y, et al. Telmisartan and N-acetylcysteine suppress group V secretory phospholipase A2 expression in TNFα-stimulated human endothelial cells and reduce associated atherogenicity. J Cardiovasc Pharmacol. 2012;60:367–74.

    Article  CAS  PubMed  Google Scholar 

  8. Muñoz NM, Desai A, Meliton LN, Meliton AY, Zhou T, Leff AR, et al. Group V phospholipase A2 increases pulmonary endothelial permeability through direct hydrolysis of the cell membrane. Pulm Circ. 2012;2:182–92.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  9. Rosenson RS, Gelb MH. Secretory phospholipase A2: a multifaceted family of proatherogenic enzymes. Curr Cardiol Rep. 2009;11:445–51.

    Article  PubMed  Google Scholar 

  10. Balestrieri B, Arm JP. Group V sPLA2: classical and novel functions. Biochim Biophys Acta. 2006;1761:1280–8.

    Article  CAS  PubMed  Google Scholar 

  11. Degousee N, Ghomashchi F, Stefanski E, Singer A, Smart BP, Borregaard N, et al. V, and X phospholipases A2s in human neutrophils: role in eicosanoid production and gram-negative bacterial phospholipid hydrolysis. J Biol Chem. 2002;277:5061–73.

    Article  CAS  PubMed  Google Scholar 

  12. Kim KP, Rafter JD, Bittova L, Han SK, Snitko Y, Munoz NM, et al. Mechanism of human group V phospholipase A2 (PLA2)-induced leukotriene biosynthesis in human neutrophils. A potential role of heparan sulfate binding in PLA2 internalization and degradation. J Biol Chem. 2001;276:11126–34.

    Article  CAS  PubMed  Google Scholar 

  13. Rubio JM, Rodríguez JP, Gil-de-Gómez L, Guijas C, Balboa MA, Balsinde J. Group V Secreted Phospholipase A2 Is Upregulated by IL-4 in Human Macrophages and Mediates Phagocytosis via Hydrolysis of Ethanolamine Phospholipids. J Immunol. 2015;194:3327–39.

    Article  CAS  PubMed  Google Scholar 

  14. Boilard E, Lai Y, Larabee K, Balestrieri B, Ghomashchi F, Fujioka D, et al. A novel anti-inflammatory role for secretory phospholipase A2 in immune complex-mediated arthritis. EMBO Mol Med. 2010;2:172–87.

    Article  PubMed  PubMed Central  Google Scholar 

  15. Murakami M, Sato H, Miki Y, Yamamoto K, Taketomi Y. A New Era of Secreted Phospholipase A2 (sPLA2). J Lipid Res. 2015;56:1248–61.

    CAS  PubMed  PubMed Central  Google Scholar 

  16. Ito M, Ishikawa Y, Kiguchi H, Komiyama K, Murakami M, Kudo I, et al. Distribution of type V secretory phospholipase A2 expression in human hepatocytes damaged by liver disease. J Gastroenterol Hepatol. 2004;19:1140–9.

    Article  CAS  PubMed  Google Scholar 

  17. Masuda S, Murakami M, Ishikawa Y, Ishii T, Kudo I. Diverse cellular localizations of secretory phospholipase A2 enzymes in several human tissues. Biochim Biophys Acta. 2005;1736:200–10.

    Article  CAS  PubMed  Google Scholar 

  18. Scott KF, Sajinovic M, Hein J, Nixdorf S, Galettis P, Liauw W, et al. Emerging roles for phospholipase A2 enzymes in cancer. Biochimie. 2010;92:601–10.

    Article  CAS  PubMed  Google Scholar 

  19. Quach ND, Arnold RD, Cummings BS. Secretory phospholipase A2 enzymes as pharmacological targets for treatment of disease. Biochem Pharmacol. 2014;90:338–48.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Brglez V, Lambeau G, Petan T. Secreted phospholipases A2 in cancer: diverse mechanisms of action. Biochimie. 2014;107:114–23.

    Article  CAS  PubMed  Google Scholar 

  21. Mounier CM, Wendum D, Greenspan E, Fléjou JF, Rosenberg DW, Lambeau G. Distinct expression pattern of the full set of secreted phospholipases A2 in human colorectal adenocarcinomas: sPLA2-III as a biomarker candidate. Br J Cancer. 2008;98:587–95.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  22. Fiancette R, Vincent C, Donnard M, Bordessoule D, Turlure P, Trimoreau F, et al. Genes encoding multiple forms of phospholipase A2 are expressed in immature forms of human leukemic blasts. Leukemia. 2009;23:1196–9.

    Article  CAS  PubMed  Google Scholar 

  23. Menschikowski M, Hagelgans A, Kostka H, Eisenhofer G, Siegert G. Involvement of epigenetic mechanisms in the regulation of secreted phospholipase A2 expressions in Jurkat leukemia cells. Neoplasia. 2008;10:1195–203.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  24. Gorovetz M, Baekelandt M, Berner A, Trope’ CG, Davidson B, Reich R. The clinical role of phospholipase A2 isoforms in advanced-stage ovarian carcinoma. Gynecol Oncol. 2006;103:831–40.

    Article  CAS  PubMed  Google Scholar 

  25. Herman JG, Baylin SB. Gene silencing in cancer in association with promoter hypermethylation. N Engl J Med. 2003;349:2042–54.

    Article  CAS  PubMed  Google Scholar 

  26. Toiyama Y, Okugawa Y, Goel A. DNA methylation and microRNA biomarkers for noninvasive detection of gastric and colorectal cancer. Biochem Biophys Res Commun. 2014;455:43–57.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  27. Menschikowski M, Hagelgans A, Gussakovsky E, Kostka H, Paley EL, Siegert G. Differential expression of secretory phospholipases A2 in normal and malignant prostate cell lines: regulation by cytokines, cell signaling pathways, and epigenetic mechanisms. Neoplasia. 2008;10:279–86.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  28. Ganesan K, Ivanova T, Wu Y, Rajasegaran V, Wu J, Lee MH, et al. Inhibition of gastric cancer invasion and metastasis by PLA2G2A, a novel beta-catenin/TCF target gene. Cancer Res. 2008;68:4277–86.

    Article  CAS  PubMed  Google Scholar 

  29. Brglez V, Pucer A, Pungerčar J, Lambeau G, Petan T. Secreted phospholipases A2 are differentially expressed and epigenetically silenced in human mammary cancer cells. Biochem Biophys Res Commun. 2014;445:230–5.

    Article  CAS  PubMed  Google Scholar 

  30. Lacroux C, Vilette D, Fernández-Borges N, Litaise C, Lugan S, Morel N, et al. Prionemia and leukocyte-platelet-associated infectivity in sheep transmissible spongiform encephalopathy models. J Virol. 2012;86:2056–66.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  31. McCurdy RD, McGrath JJ, Mackay-Sim A. Validation of the comparative quantification method of real-time PCR analysis and a cautionary tale of housekeeping gene selection. Gene Ther Mol Biol. 2008;12:15–24.

    CAS  Google Scholar 

  32. Li LC, Dahiya R. MethPrimer: designing primers for methylation PCRs. Bioinformatics. 2002;18:1427–31.

    Article  CAS  PubMed  Google Scholar 

  33. Farré D, Roset R, Huerta M, Adsuara JE, Roselló L, Albà MM, et al. Identification of patterns in biological sequences at the ALGGEN server: PROMO and MALGEN. Nucleic Acids Res. 2003;31:3651–3.

    Article  PubMed  PubMed Central  Google Scholar 

  34. Rhodes DR, Kalyana-Sundaram S, Mahavisno V, Varambally R, Yu J, Briggs BB, et al. Oncomine 3.0: genes, pathways, and networks in a collection of 18,000 cancer gene expression profiles. Neoplasia. 2007;9:166–80.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  35. Ben-Hattar J, Jiricny J. Methylation of single CpG dinucleotides within a promoter element of the Herpes simplex virus tk gene reduces its transcription in vivo. Gene. 1988;65:219–27.

    Article  CAS  PubMed  Google Scholar 

  36. Douet V, Heller MB, Le Saux O. DNA methylation and Sp1 binding determine the tissue-specific transcriptional activity of the mouse Abcc6 promoter. Biochem Biophys Res Commun. 2007;354:66–71.

    Article  CAS  PubMed  Google Scholar 

  37. Zou B, Chim CS, Zeng H, Leung SY, Yang Y, Tu SP, et al. Correlation between the single-site CpG methylation and expression silencing of the XAF1 gene in human gastric and colon cancers. Gastroenterology. 2006;131:1835–43.

    Article  CAS  PubMed  Google Scholar 

  38. Miki Y, Mukae S, Murakami M, Ishikawa Y, Ishii T, Ohki H, et al. Butyrate inhibits oral cancer cell proliferation and regulates expression of secretory phospholipase A2-X and COX-2. Anticancer Res. 2007;27:1493–502.

    CAS  PubMed  Google Scholar 

  39. Jaenisch R, Bird A. Epigenetic regulation of gene expression: how the genome integrates intrinsic and environmental signals. Nat Genet. 2003;33(Suppl):245–54.

    Article  CAS  PubMed  Google Scholar 

  40. Grivennikov SI, Greten FR, Karin M. Immunity, inflammation, and cancer. Cell. 2010;140:883–99.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  41. Menschikowski M, Platzbecker U, Hagelgans A, Vogel M, Thiede C, Schönefeldt C, et al. Aberrant methylation of the M-type phospholipase A2 receptor gene in leukemic cells. BMC Cancer. 2012;12:576.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  42. Bernard D, Vindrieux D. PLA2R1: expression and function in cancer. Biochim Biophys Acta. 1846;2014:40–4.

    Google Scholar 

  43. Surrel F, Jemel I, Boilard E, Bollinger JG, Payré C, Mounier CM, et al. Group X phospholipase A2 stimulates the proliferation of colon cancer cells by producing various lipid mediators. Mol Pharmacol. 2009;76:778–90.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  44. Pucer A, Brglez V, Payré C, Pungerčar J, Lambeau G, Petan T. Group X secreted phospholipase A2 induces lipid droplet formation and prolongs breast cancer cell survival. Mol Cancer. 2013;12:111.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  45. Martín R, Cordova C, Nieto ML. Secreted phospholipase A2-IIA-induced a phenotype of activated microglia in BV-2 cells requires epidermal growth factor receptor transactivation and proHB-EGF shedding. J Neuroinflammation. 2012;9:154.

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  46. Henderson Jr WR, Ye X, Lai Y, Ni Z, Bollinger JG, Tien YT, et al. Key role of group V secreted phospholipase A2 in Th2 cytokine and dendritic cell-driven airway hyperresponsiveness and remodeling. PLoS One. 2013;8:e56172.

    Article  CAS  PubMed  Google Scholar 

  47. Murakami M, Kambe T, Shimbara S, Higashino K, Hanasaki K, Arita H, et al. Different functional aspects of the group II subfamily (Types IIA and V) and type X secretory phospholipase A2s in regulating arachidonic acid release and prostaglandin generation. Implications of cyclooxygenase-2 induction and phospholipid scramblase-mediated cellular membrane perturbation. J Biol Chem. 1999;274:31435–44.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors are grateful to Dr. Sylke Winkler, Liane Funke and Dorit Pache for performing DNA sequencing with ABI 3.1 at the 3730 XL ABI Hitachi-sequencer (Max Planck Institute of Molecular Cell Biology and Genetics, DNA Sequencing Facility, Dresden) and to Margot Vogel and Romy Adler for their expert technical assistance.

Author information

Authors and Affiliations

  1. Institute of Clinical Chemistry and Laboratory Medicine, Carl Gustav Carus University Hospital, Technical University of Dresden, Fetscherstr. 74, D-01307, Dresden, Germany

    Mario Menschikowski, Albert Hagelgans, Brit Nacke, Carsten Jandeck & Gabriele Siegert

  2. Veterans Affairs Greater Los Angeles Healthcare System and University of California at Los Angeles, Los Angeles, CA, USA

    Olga A. Mareninova

  3. Titus Family Department of Clinical Pharmacy, USC School of Pharmacy, Los Angeles, CA, USA

    Liana Asatryan

Authors
  1. Mario Menschikowski
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Albert Hagelgans
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Brit Nacke
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Carsten Jandeck
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Olga A. Mareninova
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Liana Asatryan
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Gabriele Siegert
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Mario Menschikowski.

Ethics declarations

Conflicts of interest

None

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Figure S1

(DOC 993 kb)

Supplementary Table S1

(DOC 51 kb)

Supplementary Table S2

(DOC 35 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Menschikowski, M., Hagelgans, A., Nacke, B. et al. Epigenetic control of group V phospholipase A2 expression in human malignant cells. Tumor Biol. 37, 8097–8105 (2016). https://doi.org/10.1007/s13277-015-4670-x

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13277-015-4670-x

Keywords

Search

Navigation