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Characterization of tumor-derived mesenchymal stem cells potentially differentiating into cancer-associated fibroblasts in lung cancer

  • S. Arena
  • M. Salati
  • G. Sorgentoni
  • F. Barbisan
  • M. Orciani
Research Article
  • 128 Downloads

Abstract

Purpose

The goal of this study was to understand if mesenchymal stem cells isolated from lung tumor tissue (T-MSCs) may differentiate into cancer associated fibroblasts (CAFs), that promote neoplastic progression, angiogenesis and metastasis in the epithelial solid tumors, mimicking the tumor microenvironmental influence.

Methods

MSCs were been obtained from healthy (Control, C-MSCs) and tumor (T-MSCs) tissue of one patient who underwent a lobectomy for a lung adenocarcinoma pT1bN0. Isolated cells were characterized for the presence of molecular markers (identified by routine diagnostic characterization in differentiated tumoral cells), stemness properties, and CAF-related markers expression. Subsequently, cells were co-cultured with a lung adenocarcinoma cell line (A549 cells) to evaluate the effects on proliferation, oncogene expression and IL6 secretion.

Results

C- and T-MSCs did not present EGFR mutations unlike tumor tissue and showed a stem-like immunophenotype, characterized by the ability to differentiate towards osteo-, chondro- and adipogenic lineages. The expression of markers referred to CAFs (α-SMA, HI-1α, MMP11, VEGF, CXCL12, TGF-β1, TGF-βRII, IL6, TNFα) was significantly higher in T-MSCs than in C-MSCs. The co-cultures with A549 cells led to the over-expression of selected oncogenes and to the increase of IL6 secretion in T-MSCs but not in C-MSCs.

Conclusions

MSCs isolated from tumor tissue displayed distinct properties compared to MSCs isolated from healthy tissue, suggesting T-MSCs differentiation towards a CAF-related phenotype under the influence of the tumoral microenvironment.

Keywords

Lung cancer Mesenchymal stem cells Cancer-associated fibroblasts IL6 

Notes

Compliance with ethical standards

Conflict of interest

The authors declare that they have no conflict of interest.

Ethical approval

All procedures performed in studies involving human participants were in accordance with the ethical standards of the institutional and/or national research committee and with the 1964 Helsinki Declaration and its later amendments or comparable ethical standards.

Informed consent

Informed consent was obtained from all individual participants included in the study.

Supplementary material

12094_2018_1894_MOESM1_ESM.docx (13 kb)
Supplementary material 1 (DOCX 13 kb)
12094_2018_1894_MOESM2_ESM.docx (14 kb)
Supplementary material 2 (DOCX 13 kb)
12094_2018_1894_MOESM3_ESM.docx (12 kb)
Supplementary material 3 (DOCX 12 kb)

References

  1. 1.
    Travis WD, Brambilla E, Nicholson AG, Yatabe Y, et al. The 2015 world health organization classification of lung tumors: impact of genetic, clinical and radiologic advances since the 2004 classification. J Thorac Oncol. 2015;10:43–1260.Google Scholar
  2. 2.
    Siegel RL, Miller KD, Jemal A. Cancer statistics, 2017. CA Cancer J Clin. 2017;67:7–30.CrossRefPubMedGoogle Scholar
  3. 3.
    Liu R, Wei S, Chen J, Xu S. Mesenchymal stem cells in lung cancer tumor microenvironment: their biological properties, influence on tumor growth and therapeutic implications. Cancer Lett. 2014;353:145–52.CrossRefPubMedGoogle Scholar
  4. 4.
    Anderberg C, Pietras K. On the origin of cancer-associated fibroblasts. Cell Cycle. 2009;8:1461–2.CrossRefPubMedGoogle Scholar
  5. 5.
    Orimo A, Weinberg RA. Heterogeneity of stromal fibroblasts in tumors. Cancer Biol Ther. 2007;6:618–9.CrossRefPubMedGoogle Scholar
  6. 6.
    Navab R, Strumpf D, Bandarchi B, Zhu CQ, et al. Prognostic gene-expression signature of carcinoma-associated fibroblasts in non-small cell lung cancer. Proc Natl Acad Sci USA. 2011;108:7160–5.CrossRefPubMedPubMedCentralGoogle Scholar
  7. 7.
    Gascard P, Tlsty TD. Carcinoma-associated fibroblasts: orchestrating the composition of malignancy. Genes Dev. 2016;30:1002–19.CrossRefPubMedPubMedCentralGoogle Scholar
  8. 8.
    Siravegna G, Mussolin B, Buscarino M, Corti G, et al. Clonal evolution and resistance to EGFR blockade in the blood of colorectal cancer patients. Nat Med. 2015;21:827.CrossRefPubMedGoogle Scholar
  9. 9.
    Mariotti C, Lazzarini R, Nicolai M, Saitta A, Orsini E, Orciani M, Di Primio R. Comparative study between amniotic-fluid mesenchymal stem cells and retinal pigmented epithelium (RPE) stem cells ability to differentiate towards RPE cells. Cell Tissue Res. 2015;362:21–31.CrossRefPubMedGoogle Scholar
  10. 10.
    Dominici M, Le Blanc K, Mueller I, et al. Minimal criteria for defining multipotent mesenchymal stromal cells. The international society for cellular therapy position statement. Cytotherapy. 2006;8:315–7.CrossRefPubMedGoogle Scholar
  11. 11.
    Orciani M, Lazzarini R, Scartozzi M, Bolletta E, Mattioli-Belmonte M, Scalise A, Di Benedetto G, Di Primio R. The response of breast cancer cells to mesenchymal stem cells: a possible role of inflammation by breast implants. Plast Reconstr Surg. 2013;132:899e–910e.CrossRefPubMedGoogle Scholar
  12. 12.
    Lazzarini R, Olivieri F, Ferretti C, Mattioli-Belmonte M, Di Primio R, Orciani M. mRNAs and miRNAs profiling of mesenchymal stem cells derived from amniotic fluid and skin: the double face of the coin. Cell Tissue Res. 2014;355:121–30.CrossRefPubMedGoogle Scholar
  13. 13.
    Orciani M, Sorgentoni G, Torresetti M, Di Primio R, Di Benedetto G. MSCs and inflammation: new insights into the potential association between ALCL and breast implants. Breast Cancer Res Treat. 2016;156:65–72.CrossRefPubMedGoogle Scholar
  14. 14.
    Gazdar AF. Activating and resistance mutations of EGFR in non-small-cell lung cancer: role in clinical response to EGFR tyrosine kinase inhibitors. Oncogene. 2009;28:S24–31.CrossRefPubMedPubMedCentralGoogle Scholar
  15. 15.
    Sharma SV, Bell DW, Settleman J, Haber DA. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer. 2007;7:169–81.CrossRefPubMedGoogle Scholar
  16. 16.
    Mishra PJ, Mishra PJ, Humeniuk R, Medina DJ, Alexe G, Mesirov JP, Ganesan S, Glod JW, Banerjee D. Carcinoma-associated fibroblast-like differentiation of human mesenchymal stem cells. Cancer Res. 2008;68:4331–9.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Sharma SV, Bell DW, Settleman J, Haber DA. Epidermal growth factor receptor mutations in lung cancer. Nat Rev Cancer. 2007;7:169–81.CrossRefPubMedGoogle Scholar
  18. 18.
    Chen LY, Molina-Vila MA, Ruan SY, Su KY, et al. Coexistence of EGFR T790M mutation and common activating mutations in pretreatment non-small cell lung cancer: a systematic review and meta-analysis. Lung Cancer. 2016;94:46–53.CrossRefPubMedGoogle Scholar
  19. 19.
    Zhang W, Stabile LP, Keohavong P, Romkes M, Grandis JR, Traynor AM, Siegfried JM. Mutation and polymorphism in the EGFR-TK domain associated with lung cancer. J Thorac Oncol. 2006;1:635–47.PubMedGoogle Scholar
  20. 20.
    Lau EY, Lo J, Cheng BY, Ma MK, et al. Cancer-associated fibroblasts regulate tumor-initiating cell plasticity in hepatocellular carcinoma through c-Met/FRA1/HEY1 signaling. Cell Rep. 2016;15:1175–89.CrossRefPubMedGoogle Scholar
  21. 21.
    Li H, Fan X, Houghton J. Tumor microenvironment: the role of the tumor stroma in cancer. J Cell Biochem. 2007;101:805–15.CrossRefPubMedGoogle Scholar
  22. 22.
    Mitra AK, Zillhardt M, Hua Y, Tiwari P, Murmann AE, Peter ME, Lengyel E. Micro RNAs reprogram normal fibroblasts into cancer-associated fibroblasts in ovarian cancer. Cancer Discov. 2012;2:1100–8.CrossRefPubMedPubMedCentralGoogle Scholar
  23. 23.
    Wen S, Niu Y, Yeh S, Chang C. BM-MSCS promote prostate cancer progression via the conversion of normal fibroblasts to cancer-associated fibroblasts. Int J Oncol. 2015;47:719–27.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Orimo A, Gupta PB, Sgroi DC, Arenzana-Seisdedos F, Delaunay T, Naeem R, Carey VJ, Richardson AL, Weinberg RA. Stromal fibroblasts present in invasive human breast carcinomas promote tumor growth and angiogenesis through elevated SDF-1/CXCL12 secretion. Cell. 2005;121:335–48.CrossRefPubMedGoogle Scholar
  25. 25.
    Weekes Daniel, Kashima Takeshi G, Zandueta Carolina, Perurena Naiara, et al. Regulation of osteosarcoma cell lung metastasis by the c-Fos/AP-1 target FGFR1. Oncogene. 2016;35:2852–61.CrossRefPubMedGoogle Scholar
  26. 26.
    Schafer ZT, Brugge JS. IL-6 involvement in epithelial cancers. J Clin Invest. 2007;117:3660–3.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    Yan HQ, Huang XB, Ke SZ, Jiang YN, Zhang YH, Wang YN, Li J, Gao FG. Interleukin 6 augments lung cancer chemotherapeutic resistance via ataxia-telangiectasia mutated/NF-kappaB pathway activation. Cancer Sci. 2014;105:1220–7.CrossRefPubMedPubMedCentralGoogle Scholar
  28. 28.
    Chang CH, Hsiao CF, Yeh YM, Chang GC, et al. Circulating interleukin-6 level is a prognostic marker for survival in advanced nonsmall cell lung cancer patients treated with chemotherapy. Int J Cancer. 2013;132:1977–85.CrossRefPubMedGoogle Scholar
  29. 29.
    Shintani Y, Fujiwara A, Kimura T, Kawamura T, Funaki S, Minami M, Okumura M. IL-6 secreted from cancer-associated fibroblasts mediates chemoresistance in NSCLC by increasing epithelial-mesenchymal transition signaling. J Thorac Oncol. 2016;11:1482–92.CrossRefPubMedGoogle Scholar
  30. 30.
    Wang J, Wang B, Chu H, Yao Y. Intrinsic resistance to EGFR tyrosine kinase inhibitors in advanced non-small-cell lung cancer with activating EGFR mutations. Onco Targets Ther. 2016;9:3711–26.CrossRefPubMedPubMedCentralGoogle Scholar

Copyright information

© Federación de Sociedades Españolas de Oncología (FESEO) 2018

Authors and Affiliations

  1. 1.Department of OncologyUniversity of TorinoCandioloItaly
  2. 2.Candiolo Cancer Institute-FPO, IRCCSCandioloItaly
  3. 3.Unit of Thoracic SurgeryAOU Ospedali RiunitiAnconaItaly
  4. 4.Department of Molecular and Clinical Sciences-HistologyUniversità Politecnica delle MarcheAnconaItaly
  5. 5.Unit of PathologyAOU Ospedali RiunitiAnconaItaly

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