Applied Biochemistry and Biotechnology

, Volume 183, Issue 1, pp 444–459 | Cite as

The Inhibitory Effect of Mesenchymal Stem Cells with rAd-NK4 on Liver Cancer

  • Chao Cai
  • Lingling Hou
  • Jingsi Zhang
  • Diandian Zhao
  • Ziling Wang
  • Honggang Hu
  • Jinsheng He
  • Weijun Guan
  • Yuehui Ma
Article

Abstract

Mesenchymal stem cells (MSCs) can migrate to the tumor site and integrate into the tumor tissue. As a delivery vehicle of antitumor factors, MSCs have been tried in various tumor therapies. NK4 can both inhibit the growth, metastasis, and invasion of tumor cells induced by hepatocyte growth factor (HGF) and suppress tumor angiogenesis that is independent on HGF/cellular-mesenchymal-to-transition factor pathway. Adenovirus can directly deliver NK4 for tumor treatment but may induce immunological rejection. We combined MSCs with an adenovirus vector to deliver NK4 for liver tumor treatment. This study detected the migration of MSCs to high metastasis liver carcinoma cells MHCC-97H in vitro, investigated the inhibitory effect of rAd-NK4-MSCs on the growth and metastasis of MHCC-97H cells, further explored the inhibitory mechanism of rAd-NK4-MSCs to MHCC-97H cell metastasis, and examined the inhibitory effect of rAd-NK4-MSCs on the migration of human umbilical vein endothelial cells (HUVECs) in vitro. In this study, migration experiment was used for the potential capacity of MSCs and inhibition on migration of rAd-NK4-MSCs. Western blot was used for detecting the inhibition mechanism of rAd-NK4-MSCs to MHCC-97H cells. And, animal transplantation experiment was used for the inhibition of rAd-NK4-MSCs in vivo. MSC migration assay showed MSCs can significantly migrate to MHCC-97H cells. Co-culture results indicated that rAd-NK4-MSCs significantly inhibited the proliferation and migration of MHCC-97H cells in vitro. Western blot results proved that rAd-NK4-MSCs inhibited MHCC-97H cell migration correlating with suppressing Erk1/2 phosphorylation. HUVEC migration experiment suggested that rAd-NK4-MSCs had a potential of inhibiting tumor angiogenesis. Animal transplantation experiment showed that the tumor growth was significantly inhibited in the rAd-NK4-MSC group. Taken together, this study proved that NK4-modified MSCs had an inhibitory effect on the growth and migration of MHCC-97H cells and tumor angiogenesis, which provided a new strategy for liver tumor target therapy.

Keywords

MSCs NK4 Adenovirus vector Liver cancer cells Inhibition 

Abbreviations

5-FC

5-Fluorocytosine

bFGF

Basic fibroblast growth factor

CD

Cytosine deaminase

c-Met

Cellular-mesenchymal-to-epithelial transition factor

GCV

Ganciclovir

GMCSF

Granulocyte macrophage colony-stimulating factor

HGF

Hepatocyte growth factor

HSV-TK

Herpes simplex virus thymidine kinase

HUVECs

Human umbilical vein endothelial cells

IFN-β

Interferon-beta

IL-2

Interleukin-2

IL-15

Interleukin-15

MSCs

Mesenchymal stem cells

NK cell

Natural kill cell

rAAV

Recombinant adeno-associated virus

rMSCs

Rat MSCs

TRAIL

Tumor necrosis factor-related apoptosis-inducing ligand

VEGF

Vascular endothelial growth factor

Notes

Acknowledgements

This work was supported by the National Natural Science Foundation of China (81201762) and the National High Technology Research and Development Program (863 Program) of China (2014AA021605).

Compliance with Ethical Standards

Conflict of Interest

The authors declare that they have no competing interests.

References

  1. 1.
    Friedenstein, A. J., Chailakhjan, R. K., & Lalykina, K. S. (1970). The development of fibroblast colonies in monolayer cultures of guinea-pig bone marrow and spleen cells. Cell and Tissue Kinetics, 3, 393–403.Google Scholar
  2. 2.
    Friedenstein, A. J. (1961). Osteogenetic activity of transplanted transitional epithelium. Acta Anatomica (Basel), 45, 31–59.CrossRefGoogle Scholar
  3. 3.
    Kern, S., Eichler, H., Stoeve, J., et al. (2006). Comparative analysis of mesenchymal stem cells from bone marrow, umbilical cord blood, or adipose tissue. Stem Cells, 24, 1294–1301.CrossRefGoogle Scholar
  4. 4.
    Castillo-Melendez, M., Yawno, T., Jenkin, G., et al. (2013). Stem cell therapy to protect and repair the developing brain: a review of mechanisms of action of cord blood and amnion epithelial derived cells. Frontiers in Neuroscience, 7, 194. doi: 10.3389/fnins.2013.00194.CrossRefGoogle Scholar
  5. 5.
    Romanov, Y. A., Svintsitskaya, V. A., & Smirnov, V. N. (2003). Searching for alternative sources of postnatal human mesenchymal stem cells: candidate MSC-like cells from umbilical cord. Stem Cells, 21, 105–110.CrossRefGoogle Scholar
  6. 6.
    Troyer, D. L., & Weiss, M. L. (2008). Wharton’s jelly-derived cells are a primitive stromal cell population. Stem Cells, 26, 591–599.CrossRefGoogle Scholar
  7. 7.
    Ilancheran, S., Moodley, Y., & Manuelpillai, U. (2009). Human fetal membranes: a source of stem cells for tissue regeneration and repair? Placenta, 30, 2–10.CrossRefGoogle Scholar
  8. 8.
    Xu, F., Yang, C. C., Gomillion, C., et al. (2010). Effect of ceramide on mesenchymal stem cell differentiation toward adipocytes. Applied Biochemistry and Biotechnology, 160, 197–212.CrossRefGoogle Scholar
  9. 9.
    Cipriani, P., Di Benedetto, P., Liakouli, V., et al. (2013). Mesenchymal stem cells (MSCs) from scleroderma patients (SSc) preserve their immunomodulatory properties although senescent and normally induce T regulatory cells (Tregs) with a functional phenotype: implications for cellular-based therapy. Clinical and Experimental Immunology, 173, 195–206.CrossRefGoogle Scholar
  10. 10.
    Park, H., Cho, J. A., Kim, S. K., et al. (2008). Hyperthermia on mesenchymal stem cells (MSCs) can sensitize tumor cells to undergo cell death. International Journal of Hyperthermia, 24, 638–648.CrossRefGoogle Scholar
  11. 11.
    Hou, L., Wang, X., Zhou, Y., et al. (2014). Inhibitory effect and mechanism of mesenchymal stem cells on liver cancer cells. Tumour Biology, 35, 1239–1250.CrossRefGoogle Scholar
  12. 12.
    Zhang, B., Shan, H., Li, D., et al. (2012). The inhibitory effect of MSCs expressing TRAIL as a cellular delivery vehicle in combination with cisplatin on hepatocellular carcinoma. Cancer Biology & Therapy, 13, 1175–1184.CrossRefGoogle Scholar
  13. 13.
    Takemiya, K., Kai, H., Yasukawa, H., et al. (2010). Mesenchymal stem cell-based prostacyclin synthase gene therapy for pulmonary hypertension rats. Basic Research in Cardiology, 105, 409–417.CrossRefGoogle Scholar
  14. 14.
    Deng, Q., Zhang, Z., Feng, X., et al. (2014). TRAIL-secreting mesenchymal stem cells promote apoptosis in heat-shock-treated liver cancer cells and inhibit tumor growth in nude mice. Gene Therapy, 21, 317–327.CrossRefGoogle Scholar
  15. 15.
    Fritz, V., & Jorgensen, C. (2008). Mesenchymal stem cells: an emerging tool for cancer targeting and therapy. Current Stem Cell Research & Therapy, 3, 32–42.CrossRefGoogle Scholar
  16. 16.
    Loebinger, M. R., Eddaoudi, A., Davies, D., et al. (2009). Mesenchymal stem cell delivery of TRAIL can eliminate metastatic cancer. Cancer Research, 69, 4134–4142.CrossRefGoogle Scholar
  17. 17.
    Kang, S. G., Jeun, S. S., Lim, J. Y., et al. (2005). Cytotoxicity of rat marrow stromal cells against malignant glioma cells. Child’s Nervous System, 21, 528–538.CrossRefGoogle Scholar
  18. 18.
    Ren, C., Kumar, S., Chanda, D., et al. (2008). Cancer gene therapy using mesenchymal stem cells expressing interferon-beta in a mouse prostate cancer lung metastasis model. Gene Therapy, 15, 1446–1453.CrossRefGoogle Scholar
  19. 19.
    Wang, G. X., Zhan, Y. A., Hu, H. L., et al. (2012). Mesenchymal stem cells modified to express interferon-beta inhibit the growth of prostate cancer in a mouse model. The Journal of International Medical Research, 40, 317–327.CrossRefGoogle Scholar
  20. 20.
    Jiang, J., Wei, D., Sun, L., et al. (2014). A preliminary study on the construction of double suicide gene delivery vectors by mesenchymal stem cells and the in vitro inhibitory effects on SKOV3 cells. Oncology Reports, 31, 781–787.CrossRefGoogle Scholar
  21. 21.
    Ma, P. C., Tretiakova, M. S., Nallasura, V., et al. (2007). Downstream signalling and specific inhibition of c-MET/HGF pathway in small cell lung cancer: implications for tumour invasion. British Journal of Cancer, 97, 368–377.CrossRefGoogle Scholar
  22. 22.
    Maeda, A., Nakashiro, K., Hara, S., et al. (2006). Inactivation of AR activates HGF/c-Met system in human prostatic carcinoma cells. Biochemical and Biophysical Research Communications, 347, 1158–1165.CrossRefGoogle Scholar
  23. 23.
    Matsumura, A., Kubota, T., Taiyoh, H., et al. (2013). HGF regulates VEGF expression via the c-Met receptor downstream pathways, PI3K/Akt, MAPK and STAT3, in CT26 murine cells. International Journal of Oncology, 42, 535–542.CrossRefGoogle Scholar
  24. 24.
    Shojaei, F., Lee, J. H., Simmons, B. H., et al. (2010). HGF/c-Met acts as an alternative angiogenic pathway in sunitinib-resistant tumors. Cancer Research, 70, 10090–10100.CrossRefGoogle Scholar
  25. 25.
    de Luca, A., Arena, N., Sena, L. M., et al. (1999). Met overexpression confers HGF-dependent invasive phenotype to human thyroid carcinoma cells in vitro. Journal of Cellular Physiology, 180, 365–371.CrossRefGoogle Scholar
  26. 26.
    Date, K., Matsumoto, K., Shimura, H., et al. (1997). HGF/NK4 is a specific antagonist for pleiotrophic actions of hepatocyte growth factor. FEBS Letters, 420, 1–6.CrossRefGoogle Scholar
  27. 27.
    Kuba, K., Matsumoto, K., Date, K., et al. (2000). HGF/NK4, a four-kringle antagonist of hepatocyte growth factor, is an angiogenesis inhibitor that suppresses tumor growth and metastasis in mice. Cancer Research, 60, 6737–6743.Google Scholar
  28. 28.
    Sakai, K., Nakamura, T., Matsumoto, K., et al. (2009). Angioinhibitory action of NK4 involves impaired extracellular assembly of fibronectin mediated by perlecan-NK4 association. The Journal of Biological Chemistry, 284, 22491–22499.CrossRefGoogle Scholar
  29. 29.
    Ozen, E., Gozukizi, A., Erdal, E., et al. (2012). Heparin inhibits hepatocyte growth factor induced motility and invasion of hepatocellular carcinoma cells through early growth response protein 1. PloS One, 7, e42717. doi: 10.1371/journal.pone.0042717.CrossRefGoogle Scholar
  30. 30.
    Seo, S., & Na, K. (2011). Mesenchymal stem cell-based tissue engineering for chondrogenesis. Journal of Biomedicine & Biotechnology, 2011, 806891. doi: 10.1155/2011/806891.Google Scholar
  31. 31.
    Sharma, A. K., Hota, P. V., Matoka, D. J., et al. (2010). Urinary bladder smooth muscle regeneration utilizing bone marrow derived mesenchymal stem cell seeded elastomeric poly(1,8-octanediol-co-citrate) based thin films. Biomaterials, 31, 6207–6217.CrossRefGoogle Scholar
  32. 32.
    Lu, T., Xiong, H., Wang, K., et al. (2014). Isolation and characterization of adipose-derived mesenchymal stem cells (ADSCs) from cattle. Applied Biochemistry and Biotechnology, 174, 719–728.CrossRefGoogle Scholar
  33. 33.
    Saucier, C., Khoury, H., La, K. M., et al. (2004). The Shc adaptor protein is critical for VEGF induction by Met/HGF and ErbB2 receptors and for early onset of tumor angiogenesis. Proceedings of the National Academy of Sciences of the United States of America, 101, 2345–2350.CrossRefGoogle Scholar
  34. 34.
    Rosen, E. M., Zitnik, R. J., Elias, J. A., et al. (1993). The interaction of HGF-SF with other cytokines in tumor invasion and angiogenesis. EXS, 65, 301–310.Google Scholar
  35. 35.
    Takeuchi, S., Wang, W., Li, Q., et al. (2012). Dual inhibition of Met kinase and angiogenesis to overcome HGF-induced EGFR-TKI resistance in EGFR mutant lung cancer. The American Journal of Pathology, 181, 1034–1043.CrossRefGoogle Scholar
  36. 36.
    Davies, G., Mason, M. D., Martin, T. A., et al. (2003). The HGF/SF antagonist NK4 reverses fibroblast- and HGF-induced prostate tumor growth and angiogenesis in vivo. International Journal of Cancer, 106, 348–354.CrossRefGoogle Scholar
  37. 37.
    Deng, X. B., Xiao, L., Wu, Y., et al. (2015). Inhibition of mesothelioma cancer stem-like cells with adenovirus-mediated NK4 gene therapy. International Journal of Cancer, 137, 481–490.CrossRefGoogle Scholar
  38. 38.
    Kubota, T., Matsumura, A., Taiyoh, H., et al. (2013). Interruption of the HGF paracrine loop by NK4, an HGF antagonist, reduces VEGF expression of CT26 cells. Oncology Reports, 30, 567–572.CrossRefGoogle Scholar
  39. 39.
    Xu, C., Lin, L., Cao, G., et al. (2014). Interferon-alpha-secreting mesenchymal stem cells exert potent antitumor effect in vivo. Oncogene, 33, 5047–5052.CrossRefGoogle Scholar
  40. 40.
    Yi, B. R., Hwang, K. A., Aboody, K. S., et al. (2014). Selective antitumor effect of neural stem cells expressing cytosine deaminase and interferon-beta against ductal breast cancer cells in cellular and xenograft models. Stem Cell Research, 12, 36–48.CrossRefGoogle Scholar
  41. 41.
    Knoop, K., Schwenk, N., Schmohl, K., et al. (2015). Mesenchymal stem cell-mediated, tumor stroma-targeted radioiodine therapy of metastatic colon cancer using the sodium iodide symporter as theranostic gene. Journal of Nuclear Medicine, 56, 600–606.CrossRefGoogle Scholar
  42. 42.
    Qiao, L., Xu, Z., Zhao, T., et al. (2008). Suppression of tumorigenesis by human mesenchymal stem cells in a hepatoma model. Cell Research, 18, 500–507.CrossRefGoogle Scholar
  43. 43.
    Cao, X., Li, Y., Luo, R. Z., et al. (2015). Expression of Cystatin SN significantly correlates with recurrence, metastasis, and survival duration in surgically resected non-small cell lung cancer patients. Scientific Reports, 5, 8230. doi: 10.1038/srep08230.CrossRefGoogle Scholar
  44. 44.
    Tan, C., Qiao, F., Wei, P., et al. (2015). DIXDC1 activates the Wnt signaling pathway and promotes gastric cancer cell invasion and metastasis. Molecular Carcinogenesis, 55, 397–408.CrossRefGoogle Scholar
  45. 45.
    Marzese, D. M., Huynh, J. L., Kawas, N. P., et al. (2014). Multi-platform genome-wide analysis of melanoma progression to brain metastasis. Genom Data, 2, 150–152.CrossRefGoogle Scholar
  46. 46.
    Zhang, L., Sun, J., Wang, B., et al. (2015). MicroRNA-10b triggers the epithelial-mesenchymal transition (EMT) of laryngeal carcinoma Hep-2 cells by directly targeting the E-cadherin. Applied Biochemistry and Biotechnology, 176, 33–44.CrossRefGoogle Scholar
  47. 47.
    Marone, G., Varicchi, G., Loffredo, S., et al. (2015). Mast cells and basophils in inflammatory and tumor angiogenesis and lymphangiogenesis. European Journal of Pharmacology, 778, 146–151.CrossRefGoogle Scholar
  48. 48.
    Akrami, H., Aminzadeh, S., & Fallahi, H. (2015). Inhibitory effect of ibuprofen on tumor survival and angiogenesis in gastric cancer cell. Tumour Biology, 36, 3237–3243.CrossRefGoogle Scholar
  49. 49.
    Zeng, L., Morinibu, A., Kobayashi, M., et al. (2014). Aberrant IDH3alpha expression promotes malignant tumor growth by inducing HIF-1-mediated metabolic reprogramming and angiogenesis. Oncogene, 34, 4758–4766.CrossRefGoogle Scholar
  50. 50.
    Chougule, R. A., P, S., Salimath, B. P., et al. (2013). Buffalo colostrum β-lactoglobulin inhibits VEGF-induced angiogenesis by interacting with G protein-coupled receptor kinase. Applied Biochemistry and Biotechnology, 171, 366–381.CrossRefGoogle Scholar
  51. 51.
    Ebrahimizadeh, W., Mousavi Gargari, S. L., Javidan, Z., et al. (2015). Production of novel VHH nanobody inhibiting angiogenesis by targeting binding site of VEGF. Applied Biochemistry and Biotechnology, 176, 1985–1995.CrossRefGoogle Scholar

Copyright information

© Springer Science+Business Media New York 2017

Authors and Affiliations

  • Chao Cai
    • 1
  • Lingling Hou
    • 1
  • Jingsi Zhang
    • 1
  • Diandian Zhao
    • 1
  • Ziling Wang
    • 1
  • Honggang Hu
    • 1
  • Jinsheng He
    • 1
  • Weijun Guan
    • 2
  • Yuehui Ma
    • 2
  1. 1.College of Life Sciences and BioengineeringBeijing Jiaotong UniversityBeijingPeople’s Republic of China
  2. 2.Institute of Animal SciencesChinese Academy of Agricultural Sciences (CAAS)BeijingPeople’s Republic of China

Personalised recommendations