Cellular Changes Resulting from Forced Expression of Glypican-3 in Hepatocellular Carcinoma Cells

  • Kwack, Mi Hee (Department of Immunology, School of Medicine, Kyungpook National University) ;
  • Choi, Byung Yeol (Department of Immunology, School of Medicine, Kyungpook National University) ;
  • Sung, Young Kwan (Department of Immunology, School of Medicine, Kyungpook National University)
  • Received : 2005.11.01
  • Accepted : 2006.02.27
  • Published : 2006.04.30

Abstract

Glypican-3 (GPC3) is a member of the glypican family, which encodes cell-surface heparan-sulfate proteoglycans, and is frequently upregulated in hepatocellular carcinoma (HCC). We have recently reported that blocking endogenous GPC3 expression promotes the growth of HCC cell lines, suggesting that GPC3 plays a negative role in HCC cell proliferation. Here, we report that forced expression of GPC3 reduced the growth of HCC cells. We also found that FGF2-mediated cell proliferation was inhibited by GPC3. In addition, we observed that the adhesion of HCC cells to collagen type I and fibronectin was decreased by GPC3, whereas cellular migration and invasiveness were stimulated. Collectively, these results suggest that progression of hepatocellular carcinoma is associated with upregulation of GPC3.

Keywords

Adhesion;Glypican-3;Growth Suppression;Hepatocellular Carcinoma;Invasion;Migration

Acknowledgement

Supported by : Kyungpook National University Hospital

References

  1. Cano-Gauci, D. F., Song, H. H., Yang, H., McKerlie, C., Choo, B., et al. (1999) Glypican-3-deficient mice exhibit developmental overgrowth and some of the abnormalities typical of Simpson-Golabi-Behmel syndrome. J. Cell Biol. 146, 255-264
  2. Grisaru, S., Cano-Gauci, D., Tee, J., Filmus, J., and Rosenblum, N. D. (2001) Glypican-3 modulates BMP- and FGF-mediated effects during renal branching morphogenesis. Dev. Biol. 231, 31-46 https://doi.org/10.1006/dbio.2000.0127
  3. Hippo, Y., Watanabe, K., Watanabe, A., Midorikawa, Y., Yamamoto, S., et al. (2004) Identification of soluble NH2-terminal fragment of glypican-3 as a serological marker for early-stage hepatocellular carcinoma. Cancer Res. 64, 2418-2423 https://doi.org/10.1158/0008-5472.CAN-03-2191
  4. Midorikawa, Y., Ishikawa, S., Iwanari, H., Imamura, T., Sakamoto, H., et al. (2003) Glypican-3, overexpressed in hepatocellular carcinoma, modulates FGF2 and BMP-7 signaling. Int. J. Cancer 103, 455-465 https://doi.org/10.1002/ijc.10856
  5. Murthy, S. S., Shen, T., De Rienzo, A., Lee, W. C., Ferriola, P. C., et al. (2000) Expression of GPC3, an X-linked recessive overgrowth gene, is silenced in malignant mesothelioma. Oncogene 19, 410-416 https://doi.org/10.1038/sj.onc.1203322
  6. Nakatsura, T., Yoshitake, Y., Senju, S., Monji, M., Komori, H., et al. (2003) Glypican-3, overexpressed specifically in human hepatocellular carcinoma, is a novel tumor marker. Biochem. Biophys. Res. Commun. 306, 16-25 https://doi.org/10.1016/S0006-291X(03)00908-2
  7. Park, J. G., Lee, J. H., Kang, M. S., Park, K. J., Jeon, Y. M., et al. (1995) Characterization of cell lines established from human hepatocellular carcinoma. Int. J. Cancer 62, 276-282 https://doi.org/10.1002/ijc.2910620308
  8. Peters, M. G., Farias, E., Colombo, L., Filmus, J., Puricelli, L., et al. (2003) Inhibition of invasion and metastasis by glypican-3 in a syngeneic breast cancer model. Breast Cancer Res. Treat. 80, 221-232 https://doi.org/10.1023/A:1024549729256
  9. Pilia, G., Hughes-Benzie, R. M., MacKenzie, A., Baybayan, P., Chen, E. Y., et al. (1996) Mutations in GPC3, a glypican gene, cause the Simpson-Golabi-Behmel overgrowth syndrome. Nat. Genet. 12, 241-247 https://doi.org/10.1038/ng0396-241
  10. Sung, Y. K., Hwang, S. Y., Farooq, M., Kim, J. C., and Kim, M. K. (2003b) Growth promotion of Hep G2 hepatoma cells by antisense-mediated knockdown of glypican-3 is independent of insulin-like growth factor 2 signaling. Exp. Mol. Med. 35, 257-262 https://doi.org/10.1038/emm.2003.34
  11. Chiao, E., Fisher, P., Crisponi, L., Deiana, M., Dragatsis, I., et al. (2002) Overgrowth of a mouse model of the Simpson- Golabi-Behmel syndrome is independent of IGF signaling. Dev. Biol. 243, 185-206 https://doi.org/10.1006/dbio.2001.0554
  12. Zhu, Z. W., Friess, H., Wang, L., Abou-Shady, M., Zimmermann, A., et al. (2001) Enhanced glypican-3 expression differentiates the majority of hepatocellular carcinomas from benign hepatic disorders. Gut 48, 558-564 https://doi.org/10.1136/gut.48.4.558
  13. Sung, Y. K., Hwang, S. Y., Park, M. K., Farooq, M., Han, I. S., et al. (2003a) Glypican-3 is overexpressed in human hepatocellular carcinoma. Cancer Sci. 94, 259-262 https://doi.org/10.1111/j.1349-7006.2003.tb01430.x
  14. Xiang, Y. Y., Ladeda, V., and Filmus, J. (2001) Glypican-3 expression is silenced in human breast cancer. Oncogene 20, 7408-7412 https://doi.org/10.1038/sj.onc.1204925
  15. Kim, H., Xu, G. L., Borczuk, A. C., Busch, S., Filmus, J., et al. (2003) The heparan sulfate proteoglycan GPC3 is a potential lung tumor suppressor. Am. J. Respir. Cell Mol. Biol. 29, 694-701 https://doi.org/10.1165/rcmb.2003-0061OC
  16. Lin, H., Huber, R., Schlessinger, D., and Morin, P. J. (1999) Frequent silencing of the GPC3 gene in ovarian cancer cell lines. Cancer Res. 59, 807-810
  17. Hsu, H. C., Cheng, W., and Lai, P. L. (1997) Cloning and expression of a developmentally regulated transcript MXR7 in hepatocellular carcinoma: biological significanceand temporospatial distribution. Cancer Res. 57, 5179-5184
  18. Song, H. H., Shi, W., and Filmus, J. (1997) OCI-5/rat glypican- 3 binds to fibroblast growth factor-2 but not to insulin-like growth factor-2. J. Biol.Chem. 272, 7574-7577 https://doi.org/10.1074/jbc.272.12.7574
  19. Farooq, M., Hwang, S. Y., Park, M. K., Kim, J. C., Kim, M. K., et al. (2003) Blocking endogenous glypican-3 expression releases Hep 3B cells from G1 arrest. Mol. Cells 15, 356-360
  20. Capurro, M., Wanless, I. R., Sherman, M., Deboer, G., Shi, W., et al. (2003) Glypican-3: a novel serum and histochemical marker for hepatocellular carcinoma. Gastroenterology 125, 89-97 https://doi.org/10.1016/S0016-5085(03)00689-9
  21. Paine-Saunders, S., Viviano, B. L., Zupicich, J., Skarnes, W. C., and Saunders, S. (2000) Glypican-3 controls cellular responses to Bmp4 in limb patterning and skeletal development. Dev. Biol. 225, 179-187 https://doi.org/10.1006/dbio.2000.9831
  22. Capurro, M. I., Xiang, Y. Y., Lobe, C., and Filmus, J. (2005) Glypican-3 promotes the growth of hepatocellular carcinoma by stimulating canonical Wnt signaling. Cancer Res. 65, 6245-6254 https://doi.org/10.1158/0008-5472.CAN-04-4244