Skip to main content
SpringerLink
Log in

CCL2 secreted from cancer-associated mesothelial cells promotes peritoneal metastasis of ovarian cancer cells through the P38-MAPK pathway

  • Research Paper
  • Published:
Clinical & Experimental Metastasis Aims and scope Submit manuscript

Abstract

Epithelial ovarian cancer (EOC) is considered to secrete various factors in order to promote peritoneal dissemination through cell-to-cell interaction between cancer and mesothelial cells. We previously revealed that TGF-β secreted from EOC induces normal human peritoneal mesothelial cells (HPMCs) to differentiate into cancer-associated mesothelial cells (CAMCs). However, the relationship between tumor cells and CAMCs in EOC is still unclear. We hypothesized that CAMCs also secrete chemokines that attract cancer cells and induce peritoneal dissemination of EOC. We examined chemokines secreted from HPMCs and CAMCs by human chemokine array, and revealed that conditioned medium of CAMCs (CAMCs-CM) included many types of chemokines. The signals of CCL2 were the highest compared with other chemokines. The secretion and relative expression of CCL2 were significantly higher in CAMCs. Recombinant CCL2 promoted trans-mesothelial migration of HPMCs and the migration and invasion by EOC cells. In addition, CCL2 secreted from CAMCs promoted invasion of EOC cells. Furthermore, the neutralizing antibody of CCL2 reduced invasion by EOC. Clinical outcomes of patients whose tissue expressed higher CCR2 were significantly poorer than in patients whose tissue expression was lower. CCL2 activated the phosphorylation of p38 mitogen-activated protein kinase (MAPK). In addition, CAMCs-CM activated the p38 MAPK pathway. Phosphorylation of p38 MAPK reduced with the presence of neutralizing antibody of CCL2. In conclusion, these data indicate CCL2 in CAMCs-CM promoted the malignant potential of EOC. CCL2 plays a crucial role in the tumor microenvironment of EOC.

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
Fig. 6

Similar content being viewed by others

References

  1. Colombo N, Peiretti M, Parma G et al (2010) Newly diagnosed and relapsed epithelial ovarian carcinoma: ESMO Clinical Practice Guidelines for diagnosis, treatment and follow-up. Ann Oncol 21(Suppl 5):v23–30

    PubMed  Google Scholar 

  2. Torre LA, Islami F, Siegel RL et al (2017) Global cancer in women: burden and trends. Cancer Epidemiol Biomark Prev 26:444–457

    Google Scholar 

  3. Patient report from Gynecologic Oncology Committee (2016) Acta obstetrica et gynaecologica. Japonica 68:1117–1160

    Google Scholar 

  4. Kikkawa F, Nawa A, Ino K et al (2006) Advances in treatment of epithelial ovarian cancer. Nagoya J Med Sci 68:19–26

    PubMed  Google Scholar 

  5. Brown PO, Palmer C (2009) The preclinical natural history of serous ovarian cancer: defining the target for early detection. PLoS Med 6:e1000114

    PubMed  PubMed Central  Google Scholar 

  6. Chang LC, Huang CF, Lai MS et al (2018) Prognostic factors in epithelial ovarian cancer: a population-based study. PLoS One 13:e0194993

    PubMed  PubMed Central  Google Scholar 

  7. Pannu HK, Bristow RE, Montz FJ, Fishman EK (2003) Multidetector CT of peritoneal carcinomatosis from ovarian cancer. Radiographics 23:687–701

    PubMed  Google Scholar 

  8. Force USPST, Grossman DC, Curry SJ et al (2018) Screening for ovarian cancer: US preventive services task force recommendation statement. JAMA 319:588–594

    Google Scholar 

  9. Drabsch Y, ten Dijke P (2012) TGF-beta signalling and its role in cancer progression and metastasis. Cancer Metast Rev 31:553–568

    CAS  Google Scholar 

  10. Pickup M, Novitskiy S, Moses HL (2013) The roles of TGFbeta in the tumour microenvironment. Nat Rev Cancer 13:788–799

    CAS  PubMed  PubMed Central  Google Scholar 

  11. Yu Y, Xiao CH, Tan LD et al (2014) Cancer-associated fibroblasts induce epithelial-mesenchymal transition of breast cancer cells through paracrine TGF-beta signalling. Br J Cancer 110:724–732

    CAS  PubMed  Google Scholar 

  12. Giannoni E, Bianchini F, Masieri L et al (2010) Reciprocal activation of prostate cancer cells and cancer-associated fibroblasts stimulates epithelial-mesenchymal transition and cancer stemness. Cancer Res 70:6945–6956

    CAS  PubMed  Google Scholar 

  13. Fujikake K, Kajiyama H, Yoshihara M et al (2018) A novel mechanism of neovascularization in peritoneal dissemination via cancer-associated mesothelial cells affected by TGF-beta derived from ovarian cancer. Oncol Rep 39:193–200

    CAS  PubMed  Google Scholar 

  14. Kajiyama H, Shibata K, Ino K et al (2007) Possible involvement of SDF-1alpha/CXCR4-DPPIV axis in TGF-beta1-induced enhancement of migratory potential in human peritoneal mesothelial cells. Cell Tissue Res 330:221–229

    CAS  PubMed  Google Scholar 

  15. Sugiyama K, Kajiyama H, Shibata K et al (2014) Expression of the miR200 family of microRNAs in mesothelial cells suppresses the dissemination of ovarian cancer cells. Mol Cancer Ther 13:2081–2091

    CAS  PubMed  Google Scholar 

  16. Liu J, Chen S, Wang W et al (2016) Cancer-associated fibroblasts promote hepatocellular carcinoma metastasis through chemokine-activated hedgehog and TGF-beta pathways. Cancer Lett 379:49–59

    CAS  PubMed  Google Scholar 

  17. Worzfeld T, Pogge von Strandmann E, Huber M et al (2017) The unique molecular and cellular microenvironment of ovarian cancer. Front Oncol 7:24

    PubMed  PubMed Central  Google Scholar 

  18. Muralidhar GG, Barbolina MV (2013) Chemokine receptors in epithelial ovarian cancer. Int J Mol Sci 15:361–376

    PubMed  PubMed Central  Google Scholar 

  19. Sarvaiya PJ, Guo D, Ulasov I et al (2013) Chemokines in tumor progression metastasis. Oncotarget 4:2171–2185

    PubMed  PubMed Central  Google Scholar 

  20. EssenBioScience-IncuCyte ZOOM live cell imaging.

  21. Dobrzycka B, Mackowiak-Matejczyk B, Terlikowska KM et al (2013) Serum levels of IL-6, IL-8 and CRP as prognostic factors in epithelial ovarian cancer. Eur Cytokine Netw 24:106–113

    CAS  PubMed  Google Scholar 

  22. Popple A, Durrant LG, Spendlove I et al (2012) The chemokine, CXCL12, is an independent predictor of poor survival in ovarian cancer. Br J Cancer 106:1306–1313

    CAS  PubMed  PubMed Central  Google Scholar 

  23. O'Connor T, Borsig L, Heikenwalder M (2015) CCL2-CCR2 signaling in disease pathogenesis. Endocr Metab Immune Disord Drug Targets 15:105–118

    CAS  PubMed  Google Scholar 

  24. Li S, Lu J, Chen Y et al (2017) MCP-1-induced ERK/GSK-3beta/Snail signaling facilitates the epithelial-mesenchymal transition and promotes the migration of MCF-7 human breast carcinoma cells. Cell Mol Immunol 14:621–630

    CAS  PubMed  Google Scholar 

  25. Lin TH, Liu HH, Tsai TH et al (2013) CCL2 increases alphavbeta3 integrin expression and subsequently promotes prostate cancer migration. Biochim Biophys Acta 1830:4917–4927

    CAS  PubMed  Google Scholar 

  26. Furukawa S, Soeda S, Kiko Y et al (2013) MCP-1 promotes invasion and adhesion of human ovarian cancer cells. Anticancer Res 33:4785–4790

    CAS  PubMed  Google Scholar 

  27. Tsaur I, Rutz J, Makarevic J et al (2015) CCL2 promotes integrin-mediated adhesion of prostate cancer cells in vitro. World J Urol 33:1051–1056

    CAS  PubMed  Google Scholar 

  28. Eferl R (2013) CCL2 at the crossroad of cancer metastasis. JAKSTAT 2:e23816

    PubMed  PubMed Central  Google Scholar 

  29. Fang WB, Jokar I, Zou A et al (2012) CCL2/CCR2 chemokine signaling coordinates survival and motility of breast cancer cells through Smad3 protein- and p42/44 mitogen-activated protein kinase (MAPK)-dependent mechanisms. J Biol Chem 287:36593–36608

    CAS  PubMed  PubMed Central  Google Scholar 

  30. He S, He S, Chen CH et al (2015) The chemokine (CCL2-CCR2) signaling axis mediates perineural invasion. Mol Cancer Res 13:380–390

    CAS  PubMed  Google Scholar 

  31. Chien J, Neums L, Powell A et al (2018) Genetic evidence for early peritoneal spreading in pelvic high-grade serous cancer. Front Oncol 8:58

    PubMed  PubMed Central  Google Scholar 

  32. Pasquier J, Gosset M, Geyl C et al (2018) CCL2/CCL5 secreted by the stroma induce IL-6/PYK2 dependent chemoresistance in ovarian cancer. Mol Cancer 17:47

    PubMed  PubMed Central  Google Scholar 

  33. Solinas G, Germano G, Mantovani A, Allavena P (2009) Tumor-associated macrophages (TAM) as major players of the cancer-related inflammation. J Leukoc Biol 86:1065–1073

    CAS  PubMed  Google Scholar 

  34. Negus RP, Stamp GW, Relf MG et al (1995) The detection and localization of monocyte chemoattractant protein-1 (MCP-1) in human ovarian cancer. J Clin Invest 95:2391–2396

    CAS  PubMed  PubMed Central  Google Scholar 

  35. Hefler L, Tempfer C, Heinze G et al (1999) Monocyte chemoattractant protein-1 serum levels in ovarian cancer patients. Br J Cancer 81:855–859

    CAS  PubMed  PubMed Central  Google Scholar 

  36. Mikula-Pietrasik J, Uruski P, Szubert S et al (2016) Biochemical composition of malignant ascites determines high aggressiveness of undifferentiated ovarian tumors. Med Oncol 33:94

    PubMed  Google Scholar 

  37. Tsuyada A, Chow A, Wu J et al (2012) CCL2 mediates cross-talk between cancer cells and stromal fibroblasts that regulates breast cancer stem cells. Cancer Res 72:2768–2779

    CAS  PubMed  PubMed Central  Google Scholar 

  38. O'Hayre M, Salanga CL, Handel TM, Allen SJ (2008) Chemokines and cancer: migration, intracellular signalling and intercellular communication in the microenvironment. Biochem J 409:635–649

    CAS  PubMed  Google Scholar 

  39. Pahler JC, Tazzyman S, Erez N et al (2008) Plasticity in tumor-promoting inflammation: impairment of macrophage recruitment evokes a compensatory neutrophil response. Neoplasia 10:329–340

    CAS  PubMed  PubMed Central  Google Scholar 

  40. Green JM, Alvero AB, Kohen F, Mor G (2009) 7-(O)-Carboxymethyl daidzein conjugated to N-t-Boc-hexylenediamine: a novel compound capable of inducing cell death in epithelial ovarian cancer stem cells. Cancer Biol Ther 8:1747–1753

    CAS  PubMed  PubMed Central  Google Scholar 

  41. Puvanenthiran S, Essapen S, Seddon AM, Modjtahedi H (2016) Impact of the putative cancer stem cell markers and growth factor receptor expression on the sensitivity of ovarian cancer cells to treatment with various forms of small molecule tyrosine kinase inhibitors and cytotoxic drugs. Int J Oncol 49:1825–1838

    CAS  PubMed  PubMed Central  Google Scholar 

  42. Sun W, Li WJ, Wei FQ et al (2016) Blockade of MCP-1/CCR4 signaling-induced recruitment of activated regulatory cells evokes an antitumor immune response in head and neck squamous cell carcinoma. Oncotarget 7:37714–37727

    PubMed  PubMed Central  Google Scholar 

  43. Fader AN, Rasool N, Vaziri SA et al (2010) CCL2 expression in primary ovarian carcinoma is correlated with chemotherapy response and survival outcomes. Anticancer Res 30:4791–4798

    CAS  PubMed  Google Scholar 

  44. Saito T, Katabuchi H (2016) Annual Report of the Committee on Gynecologic Oncology, Japan Society of Obstetrics and Gynecology: patient annual report for 2013 and treatment annual report for 2008. J Obstet Gynaecol Res 42: 1069–1079

    PubMed  Google Scholar 

  45. Wolf MJ, Hoos A, Bauer J et al (2012) Endothelial CCR2 signaling induced by colon carcinoma cells enables extravasation via the JAK2-Stat5 and p38MAPK pathway. Cancer Cell 22:91–105

    CAS  PubMed  Google Scholar 

  46. Mellado M, Rodriguez-Frade JM, Aragay A et al (1998) The chemokine monocyte chemotactic protein 1 triggers Janus kinase 2 activation and tyrosine phosphorylation of the CCR2B receptor. J Immunol 161:805–813

    CAS  PubMed  Google Scholar 

  47. Zhuang H, Cao G, Kou C, Liu T (2018) CCL2/CCR2 axis induces hepatocellular carcinoma invasion and epithelial-mesenchymal transition in vitro through activation of the Hedgehog pathway. Oncol Rep 39:21–30

    CAS  PubMed  Google Scholar 

Download references

Acknowledgements

We sincerely thank Mr. Y. Koya and Y. Yamakita (Bell Research Center, Department of Obstetrics and Gynecology Collaborative Research, Nagoya University Graduate School of Medicine) for various forms of technical support.

Funding

This study was supported by JSPS (Japan Society for the Promotion of Science) KAKENHI Grants-in-Aid for Scientific Research: Grant Numbers 17H04338 and 16K15704.

Author information

Authors and Affiliations

  1. Department of Obstetrics and Gynecology, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya, 466-8550, Japan

    Hiroaki Yasui, Hiroaki Kajiyama, Satoshi Tamauchi, Shiro Suzuki, Yang Peng, Nobuhisa Yoshikawa, Mai Sugiyama, Kae Nakamura & Fumitaka Kikkawa

  2. Division of Clinical Genetics, Department of Laboratory Medicine, Lund University, BMC C13, 22184, Lund, Sweden

    Hiroaki Yasui

Authors
  1. Hiroaki Yasui
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Hiroaki Kajiyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Satoshi Tamauchi
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Shiro Suzuki
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Yang Peng
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Nobuhisa Yoshikawa
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Mai Sugiyama
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Kae Nakamura
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Fumitaka Kikkawa
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Hiroaki Kajiyama.

Ethics declarations

Conflict of interest

All authors declare that they have no conflict of interest.

Additional information

Publisher's Note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

Table S1 Clinical characteristics of patient ascites (DOCX 15 kb)

Table S2 Clinical characteristics of patients on survival analyses (DOCX 18 kb)

10585_2019_9993_MOESM3_ESM.jpg

Fig. S1 The expressions of CCR2 in EOC cell lines. The protein levels of SKOV-3, A-2780, and OVCAR-3 were high. However, that of ES-2 was low (JPG 76 kb)

10585_2019_9993_MOESM4_ESM.jpg

Fig. S2 Proliferation assay to assess the effect of CCL2 for the cell proliferation. There was no correlation between CCL2 and the cell proliferation (JPG 224 kb)

10585_2019_9993_MOESM5_ESM.jpg

Fig. S3 Survival analyses on stratification by each histological type of EOC. A: overall survival (serous carcinoma), B: overall survival (clear-cell carcinoma), C: progression-free survival (serous carcinoma), D: progression-free survival (clear-cell carcinoma) (JPG 430 kb)

10585_2019_9993_MOESM6_ESM.jpg

Fig. S4 Schematic summary of the current study. Enhanced production of CCL2 from CAMCs is crucial for the development of peritoneal carcinomatosis of EOC through the adhesion-, migration-, and invasion-promoting effects of tumor cells. These findings indicate the scheme of the auto-stimulatory triangle axis among cancer, ascites, and the mesothelium (JPG 528 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Yasui, H., Kajiyama, H., Tamauchi, S. et al. CCL2 secreted from cancer-associated mesothelial cells promotes peritoneal metastasis of ovarian cancer cells through the P38-MAPK pathway. Clin Exp Metastasis 37, 145–158 (2020). https://doi.org/10.1007/s10585-019-09993-y

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s10585-019-09993-y

Keywords

Search

Navigation