Skip to main content

Advertisement

SpringerLink
Log in

Inhibitory effect and mechanism of mesenchymal stem cells on liver cancer cells

  • Research Article
  • Published:
Tumor Biology

Abstract

Mesenchymal stem cells (MSCs), with their capacity for self-renewal and differentiation into various cell types, are important seed cells for stem cell therapy. MSCs exhibit potent pathotropic migratory properties that make them attractive for use in tumor prevention and therapy. However, little is known about the underlying molecular mechanisms that link MSCs to the targeted tumor cells. This study investigated the inhibitory effect and mechanism of MSCs on human hepatoma HepG2 cells using co-culture and conditioned medium system and animal transplantation model. The HepG2 cells were co-cultured with MSCs or treated with conditional media derived from MSCs cultures in vitro. Results of methylthiazolyldiphenyl tetrazolium assay and flow cytometric assay showed that the proliferation and apoptosis of HepG2 cells decreased and increased, respectively. Reverse transcription polymerase chain reaction analysis showed that the expression levels of bcl-2, c-Myc, β-catenin, and survivin were downregulated. The results of enzyme-linked immunosorbent assay and Western blot proved that MSCs secreted Dkk-1 to inhibit the expression of Wnt signaling pathway-related factors (bcl-2, c-Myc, β-catenin, and survivin) in tumor cells, consequently inhibiting the proliferation and promoting the apoptosis of HepG2 cells. Animal transplantation experiment showed that tumor growth was significantly inhibited when HepG2 cells were co-injected with MSCs into nude mice. These results suggested that MSCs inhibited the growth and promoted the apoptosis of HepG2 cells in a dose-dependent manner. This study provided a new approach and experimental basis for cancer therapy. This study also proved that the Wnt signaling pathway may have a function in MSC-mediated tumor cell inhibition.

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
Fig. 7
Fig. 8
Fig. 9
Fig. 10

Similar content being viewed by others

References

  1. Molloy AP, Martin FT, Dwyer RM, Griffin TP, Murphy M, Barry FP, et al. Mesenchymal stem cell secretion of chemokines during differentiation into osteoblasts, and their potential role in mediating interactions with breast cancer cells. Int J Cancer. 2009;124(2):326–32.

    Article  CAS  PubMed  Google Scholar 

  2. Djouad F, Bouffi C, Ghannam S, Noel D, Jorgensen C. Mesenchymal stem cells: innovative therapeutic tools for rheumatic diseases. Nat Rev Rheumatol. 2009;5(7):392–9.

    Article  CAS  PubMed  Google Scholar 

  3. Khakoo AY, Pati S, Anderson SA, Reid W, Elshal MF, Rovira II, et al. Human mesenchymal stem cells exert potent antitumorigenic effects in a model of Kaposi's sarcoma. J Exp Med. 2006;203(5):1235–47.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  4. Voswinkel J, Francois S, Simon JM, Benderitter M, Gorin NC, Mohty M, et al. Use of mesenchymal stem cells (MSC) in chronic inflammatory fistulizing and fibrotic diseases: a comprehensive review. Clin Rev Allergy Immunol. 2013;45(2):180–92.

    Article  PubMed  Google Scholar 

  5. Ringden O, Uzunel M, Rasmusson I, Remberger M, Sundberg B, Lonnies H, et al. Mesenchymal stem cells for treatment of therapy-resistant graft-versus-host disease. Transplantation. 2006;81(10):1390–7.

    Article  PubMed  Google Scholar 

  6. Maestroni GJ, Hertens E, Galli P. Factor(s) from nonmacrophage bone marrow stromal cells inhibit Lewis lung carcinoma and B16 melanoma growth in mice. Cell Mol Life Sci. 1999;55(4):663–7.

    Article  CAS  PubMed  Google Scholar 

  7. Deng W, Han Q, Liao L, You S, Deng H, Zhao RC. Effects of allogeneic bone marrow-derived mesenchymal stem cells on T and B lymphocytes from BXSB mice. DNA Cell Biol. 2005;24(7):458–63.

    Article  CAS  PubMed  Google Scholar 

  8. Pisati F, Belicchi M, Acerbi F, Marchesi C, Giussani C, Gavina M, et al. Effect of human skin-derived stem cells on vessel architecture, tumor growth, and tumor invasion in brain tumor animal models. Cancer Res. 2007;67(7):3054–63.

    Article  CAS  PubMed  Google Scholar 

  9. Chen X, Lin X, Zhao J, Shi W, Zhang H, Wang Y, et al. A tumor-selective biotherapy with prolonged impact on established metastases based on cytokine gene-engineered MSCs. Mol Ther. 2008;16(4):749–56.

    Article  CAS  PubMed  Google Scholar 

  10. Kang SG, Jeun SS, Lim JY, Yoo DS, Huh PW, Cho KS, et al. Cytotoxicity of rat marrow stromal cells against malignant glioma cells. Childs Nerv Syst. 2005;21(7):528–38.

    Article  PubMed  Google Scholar 

  11. Reya T, Morrison SJ, Clarke MF, Weissman IL. Stem cells, cancer, and cancer stem cells. Nature. 2001;414(6859):105–11.

    Article  CAS  PubMed  Google Scholar 

  12. Menon LG, Picinich S, Koneru R, Gao H, Lin SY, Koneru M, et al. Differential gene expression associated with migration of mesenchymal stem cells to conditioned medium from tumor cells or bone marrow cells. Stem Cells. 2007;25(2):520–8.

    Article  CAS  PubMed  Google Scholar 

  13. Willert K, Jones KA. Wnt signaling: is the party in the nucleus? Genes Dev. 2006;20(11):1394–404.

    Article  CAS  PubMed  Google Scholar 

  14. Reya T, Clevers H. Wnt signalling in stem cells and cancer. Nature. 2005;434(7035):843–50.

    Article  CAS  PubMed  Google Scholar 

  15. Raida M, Heymann AC, Gunther C, Niederwieser D. Role of bone morphogenetic protein 2 in the crosstalk between endothelial progenitor cells and mesenchymal stem cells. Int J Mol Med. 2006;18(4):735–9.

    CAS  PubMed  Google Scholar 

  16. Miele L, Miao H, Nickoloff BJ. NOTCH signaling as a novel cancer therapeutic target. Curr Cancer Drug Targets. 2006;6(4):313–23.

    Article  CAS  PubMed  Google Scholar 

  17. Lindvall C, Evans NC, Zylstra CR, Li Y, Alexander CM, Williams BO. The Wnt signaling receptor Lrp5 is required for mammary ductal stem cell activity and Wnt1-induced tumorigenesis. J Biol Chem. 2006;281(46):35081–7.

    Article  CAS  PubMed  Google Scholar 

  18. Androutsellis-Theotokis A, Leker RR, Soldner F, Hoeppner DJ, Ravin R, Poser SW, et al. Notch signalling regulates stem cell numbers in vitro and in vivo. Nature. 2006;442(7104):823–6.

    Article  CAS  PubMed  Google Scholar 

  19. Gregory CA, Singh H, Perry AS, Prockop DJ. The Wnt signaling inhibitor dickkopf-1 is required for reentry into the cell cycle of human adult stem cells from bone marrow. J Biol Chem. 2003;278(30):28067–78.

    Article  CAS  PubMed  Google Scholar 

  20. Gaspar C, Fodde R. APC dosage effects in tumorigenesis and stem cell differentiation. Int J Dev Biol. 2004;48(5–6):377–86.

    Article  CAS  PubMed  Google Scholar 

  21. Chen G, Shukeir N, Potti A, Sircar K, Aprikian A, Goltzman D, et al. Up-regulation of Wnt-1 and beta-catenin production in patients with advanced metastatic prostate carcinoma: potential pathogenetic and prognostic implications. Cancer. 2004;101(6):1345–56.

    Article  CAS  PubMed  Google Scholar 

  22. Neth P, Ciccarella M, Egea V, Hoelters J, Jochum M, Ries C. Wnt signaling regulates the invasion capacity of human mesenchymal stem cells. Stem Cells. 2006;24(8):1892–903.

    Article  CAS  PubMed  Google Scholar 

  23. Yang F, Zeng Q, Yu G, Li S, Wang CY. Wnt/beta-catenin signaling inhibits death receptor-mediated apoptosis and promotes invasive growth of HNSCC. Cell Signal. 2006;18(5):679–87.

    Article  CAS  PubMed  Google Scholar 

  24. Baek SH, Kioussi C, Briata P, Wang D, Nguyen HD, Ohgi KA, et al. Regulated subset of G1 growth-control genes in response to derepression by the Wnt pathway. Proc Natl Acad Sci U S A. 2003;100(6):3245–50.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  25. Muncan V, Sansom OJ, Tertoolen L, Phesse TJ, Begthel H, Sancho E, et al. Rapid loss of intestinal crypts upon conditional deletion of the Wnt/Tcf-4 target gene c-Myc. Mol Cell Biol. 2006;26(22):8418–26.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  26. Joubert A, Bianchi P, Maritz C, Joubert F. Influence of prostaglandin A2 on Bax, Bcl-2 and PCNA expression in MCF-7 cells. Biomed Res. 2006;27(4):157–62.

    Article  CAS  PubMed  Google Scholar 

  27. Yang B, Guo H, Zhang Y, Chen L, Ying D, Dong S. MicroRNA-145 regulates chondrogenic differentiation of mesenchymal stem cells by targeting Sox9. PLoS ONE. 2011;6(7):e21679.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  28. Dwyer RM, Khan S, Barry FP, O'Brien T, Kerin MJ. Advances in mesenchymal stem cell-mediated gene therapy for cancer. Stem Cell Res Ther. 2010;1(3):25.

    Article  PubMed Central  PubMed  Google Scholar 

  29. Itakura S, Asari S, Rawson J, Ito T, Todorov I, Liu CP, et al. Mesenchymal stem cells facilitate the induction of mixed hematopoietic chimerism and islet allograft tolerance without GVHD in the rat. Am J Transplant. 2007;7(2):336–46.

    Article  CAS  PubMed  Google Scholar 

  30. Ramasamy R, Lam EW, Soeiro I, Tisato V, Bonnet D, Dazzi F. Mesenchymal stem cells inhibit proliferation and apoptosis of tumor cells: impact on in vivo tumor growth. Leukemia. 2007;21(2):304–10.

    Article  CAS  PubMed  Google Scholar 

  31. Ohlsson LB, Varas L, Kjellman C, Edvardsen K, Lindvall M. Mesenchymal progenitor cell-mediated inhibition of tumor growth in vivo and in vitro in gelatin matrix. Exp Mol Pathol. 2003;75(3):248–55.

    Article  CAS  PubMed  Google Scholar 

  32. Wang J, Shou J, Chen X. Dickkopf-1, an inhibitor of the Wnt signaling pathway, is induced by p53. Oncogene. 2000;19(14):1843–8.

    Article  CAS  PubMed  Google Scholar 

  33. Qiao L, Xu ZL, Zhao TJ, Ye LH, Zhang XD. Dkk-1 secreted by mesenchymal stem cells inhibits growth of breast cancer cells via depression of Wnt signalling. Cancer Lett. 2008;269(1):67–77.

    Article  CAS  PubMed  Google Scholar 

  34. Zhang L, Xiang J, Li G. The uncertain role of unmodified mesenchymal stem cells in tumor progression: what master switch? Stem Cell Res Ther. 2013;4(2):22.

    Article  CAS  PubMed  Google Scholar 

  35. Shah K. Mesenchymal stem cells engineered for cancer therapy. Adv Drug Deliv Rev. 2012;64(8):739–48.

    Article  CAS  PubMed Central  PubMed  Google Scholar 

  36. Ishii G, Sangai T, Oda T, Aoyagi Y, Hasebe T, Kanomata N, et al. Bone-marrow-derived myofibroblasts contribute to the cancer-induced stromal reaction. Biochem Biophys Res Commun. 2003;309(1):232–40.

    Article  CAS  PubMed  Google Scholar 

  37. Muehlberg FL, Song YH, Krohn A, Pinilla SP, Droll LH, Leng X, et al. Tissue-resident stem cells promote breast cancer growth and metastasis. Carcinogenesis. 2009;30(4):589–97.

    Article  CAS  PubMed  Google Scholar 

  38. Serakinci N, Christensen R, Fahrioglu U, Sorensen FB, Dagnaes-Hansen F, Hajek M, et al. Mesenchymal stem cells as therapeutic delivery vehicles targeting tumor stroma. Cancer Biother Radiopharm. 2011;26(6):767–73.

    Article  CAS  PubMed  Google Scholar 

  39. Otsu K, Das S, Houser SD, Quadri SK, Bhattacharya S, Bhattacharya J. Concentration-dependent inhibition of angiogenesis by mesenchymal stem cells. Blood. 2009;113(18):4197–205.

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgment

This work was supported by the Fundamental Research Funds for the Central Universities (2011JBM294), the National Natural Science Foundation of China (81201762), Foundation of State Key Laboratory Cultivation Base for the Chemistry and Molecular Engineering of Medicinal Resources, Ministry of Science and Technology of China (CMEMR2012-B07), and the National Basic Research Program of China (973 Program 2009CB521704). The authors thank Dr. Juan Du and Dr. Honggang Hu for critical reading of the manuscript.

Conflicts of Interest

None

Author information

Authors and Affiliations

  1. College of Life Sciences and Bioengineering, Beijing Jiaotong University, Beijing, 100044, People’s Republic of China

    Lingling Hou, Xiaoyu Wang, Yaqiong Zhou, Haibin Ma, Ziling Wang, Jinsheng He & Honggang Hu

  2. Institute of Animal Sciences, Chinese Academy of Agricultural Sciences (CAAS), Beijing, 100193, People’s Republic of China

    Weijun Guan & Yuehui Ma

Authors
  1. Lingling Hou
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Xiaoyu Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Yaqiong Zhou
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Haibin Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Ziling Wang
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Jinsheng He
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Honggang Hu
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Weijun Guan
    View author publications

    You can also search for this author in PubMed Google Scholar

  9. Yuehui Ma
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding authors

Correspondence to Lingling Hou, Weijun Guan or Yuehui Ma.

Additional information

Lingling Hou and Xiaoyu Wang contributed equally to this work.

Electronic supplementary material

Below is the link to the electronic supplementary material.

Supplementary Fig. 1

Apoptosis morphology of HepG2 induced by MSCs. (A) Cell morphology of the MSC:HepG2=5:1 group. HepG2 cell volume was reduced, cytoplasm density was increased, nuclear membrane and nucleolus were fragmented, and membrane structure was still intact, as indicated by an arrow. (B) Cell morphology of the MSC:HepG2=1:1 group, a small number of HepG2 cells showed visible cytoplasm agglutination and became round, as indicated by an arrow. (C) Cell morphology of the MSC:HepG2=1:5 group, apoptosis of HepG2 cells was not obvious, most cells were still in normal shape. (200×) (JPEG 32 kb)

High resolution (TIFF 1306 kb)

Supplementary Fig. 2

mRNA expression levels of Bcl-2, c-Myc, β-catenin, and survivin in HepG2 cells treated with MSC-conditioned media at different treatment times. Lane 1, HepG2 control group at 24 h; Lane 2, HepG2 cells were treated with 80 % MSC-conditioned media for 24 h; Lane 3, HepG2 cells were treated with 80 % HEK 293-conditioned media for 24 h; Lane 4, HepG2 control group at 48 h; Lane 5, HepG2 cells were treated with 80 % MSC-conditioned media for 48 h; Lane 6, HepG2 cells were treated with 80 % HEK 293-conditioned media for 48 h; Lane 7, HepG2 control group at 72 h; Lane 8, HepG2 cells were treated with 80 % MSC-conditioned media for 72 h; and Lane 9, HepG2 cells were treated with 80 % HEK 293-conditioned media for 72 h (JPEG 18 kb)

High resolution (TIFF 2687 kb)

Supplementary Fig. 3

mRNA expression levels of bcl-2, c-Myc, β-catenin, and survivin in HepG2 cells co-cultured with MSCs. (A) HepG2:MSC=5:1 co-culture groups; (B) HepG2:MSC=1:1 co-culture groups; (C) HepG2:MSC=1:5 co-culture groups; Lane 1, HepG2 control group at 24 h; Lane 2, HepG2 cells were co-cultured with MSCs for 24 h; Lane 3, HepG2 cells were co-cultured with HEK 293 cells for 24 h; Lane 4, HepG2 control group at 48 h; Lane 5, HepG2 cells were co-cultured with MSCs for 48 h; Lane 6, HepG2 cells were co-cultured with HEK 293 cells for 48 h; Lane 7, HepG2 control group at 72 h; Lane 8, HepG2 cells were co-cultured with MSCs for 72 h; and Lane 9, HepG2 cells were co-cultured with HEK 293 cells for 72 h (JPEG 55 kb)

High resolution (TIFF 7781 kb)

Supplementary Fig. 4

Pathological examination of tumor tissues (200×). Rich and dense tumor cells were detected in the sections of the HepG2 and HEK 293 control groups (Figs. 14A, 14D, 14E, 14F, and 14H); less tumor cells and more connective tissues in the MSC section: HepG2=1:1 and MSC:HepG2=2:1 groups than the HepG2 and HEK 293 control groups (Figs. 14B and 14C) (JPEG 110 kb)

High resolution (TIFF 4800 kb)

Rights and permissions

Reprints and permissions

About this article

Cite this article

Hou, L., Wang, X., Zhou, Y. et al. Inhibitory effect and mechanism of mesenchymal stem cells on liver cancer cells. Tumor Biol. 35, 1239–1250 (2014). https://doi.org/10.1007/s13277-013-1165-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13277-013-1165-5

Keywords

Search

Navigation