Overexpression of GPC3 inhibits hepatocellular carcinoma cell proliferation and invasion through induction of apoptosis

Pan,Zhijian; Chen,Chunzhou; Long,Haocheng; Lei,Changjiang; Tang,Gang; Li,Lei; Feng,Jiarui; Chen,Feixiang

doi:10.3892/mmr.2013.1279

Overexpression of GPC3 inhibits hepatocellular carcinoma cell proliferation and invasion through induction of apoptosis

Authors:
- Zhijian Pan
- Chunzhou Chen
- Haocheng Long
- Changjiang Lei
- Gang Tang
- Lei Li
- Jiarui Feng
- Feixiang Chen
View Affiliations

Affiliations: Second Department of General Surgery, Fifth Hospital of Wuhan, Hubei 430050, P.R. China
Published online on: January 18, 2013 https://doi.org/10.3892/mmr.2013.1279
Pages: 969-974

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Glypican‑3 (GPC3) is a membrane heparan sulfate proteoglycan involved in cell proliferation, differentiation, adhesion, migration and the development of the majority of mesodermal tissues and organs. GPC3 has been found to be important for the occurrence and development of hepatocellular carcinoma (HCC). Therefore, it may be suitable for use as a novel molecular marker for the diagnosis of primary liver cancer. In the present study, the role of GPC3 in the occurrence and development of HCC was determined. GPC3 recombinant vector was transfected into two HCC cell lines, Huh7 and SK‑HEP‑1, to upregulate the expression of GPC3 and examine changes in the biological behavior of the cells. Results indicate that overexpression of GPC3 in Huh7 and SK‑HEP‑1 cells effectively inhibited cell proliferation and cell invasion through induction of apoptosis. However, cotreatment of the cells with insulin‑like growth factor 2 (IGF2) and fibroblast growth factor 2 (FGF2) was found by Annexin V‑PI flow cytometric analysis to significantly inhibit the apoptotic cell death induced by GPC3 overexpression. These observations indicate that GPC3 may act as a negative regulator of IGF2 and FGF2 pathways. Taken together, these results demonstrate that overexpression of GPC3 inhibits the occurrence and development of HCC.

View Figures

View References

1	El-Serag HB and Rudolph KL: Hepatocellular carcinoma: epidemiology and molecular carcinogenesis. Gastroenterology. 132:2557–2576. 2007. View Article : Google Scholar : PubMed/NCBI
2	Yuen MF and Lai CL: Serological markers of liver cancer. Best Pract Res Clin Gastroenterol. 19:91–99. 2005. View Article : Google Scholar : PubMed/NCBI
3	Liu H, Li P, Zhai Y, et al: Diagnostic value of glypican-3 in serum and liver for primary hepatocellular carcinoma. World J Gastroenterol. 16:4410–4415. 2010. View Article : Google Scholar : PubMed/NCBI
4	Akutsu N, Yamamoto H, Sasaki S, et al: Association of glypican-3 expression with growth signaling molecules in hepatocellular carcinoma. World J Gastroenterol. 16:3521–3528. 2010. View Article : Google Scholar : PubMed/NCBI
5	Yan B, Wei JJ, Qian YM, et al: Expression and clinicopathologic significance of glypican 3 in hepatocellular carcinoma. Ann Diagn Pathol. 15:162–169. 2011. View Article : Google Scholar : PubMed/NCBI
6	Iglesias BV, Centeno G, Pascuccelli H, et al: Expression pattern of glypican-3 (GPC3) during human embryonic and fetal development. Histol Histopathol. 23:1333–1340. 2008.PubMed/NCBI
7	Hippo Y, Watanabe K, Watanabe A, et al: Identification of soluble NH2-terminal fragment of glypican-3 as a serological marker for early-stage hepatocellular carcinoma. Cancer Res. 64:2418–2423. 2004. View Article : Google Scholar : PubMed/NCBI
8	Capurro MI, Xiang YY, Lobe C and Filmus J: Glypican-3 promotes the growth of hepatocellular carcinoma by stimulating canonical Wnt signaling. Cancer Res. 65:6245–6254. 2005. View Article : Google Scholar : PubMed/NCBI
9	Capurro MI, Xu P, Shi W, Li F, Jia A and Filmus J: Glypican-3 inhibits Hedgehog signaling during development by competing with patched for Hedgehog binding. Dev Cell. 14:700–711. 2008. View Article : Google Scholar : PubMed/NCBI
10	Midorikawa Y, Ishikawa S, Iwanari H, et al: Glypican-3, overexpressed in hepatocellular carcinoma, modulates FGF2 and BMP-7 signaling. Int J Cancer. 103:455–465. 2003. View Article : Google Scholar : PubMed/NCBI
11	Cheng W, Tseng CJ, Lin TT, et al: Glypican-3-mediated oncogenesis involves the insulin-like growth factor-signaling pathway. Carcinogenesis. 29:1319–1326. 2008. View Article : Google Scholar : PubMed/NCBI
12	Sakurai M, Shibata K, Umezu T, et al: Growth-suppressing function of glypican-3 (GPC3) via insulin like growth factor II (IGF-II) signaling pathway in ovarian clear cell carcinoma cells. Gynecol Oncol. 119:332–336. 2010. View Article : Google Scholar : PubMed/NCBI
13	Sun CK, Chua MS, He J and So SK: Suppression of glypican 3 inhibits growth of hepatocellular carcinoma cells through up-regulation of TGF-β2. Neoplasia. 13:735–747. 2011.PubMed/NCBI
14	Kwack MH, Choi BY and Sung YK: Cellular changes resulting from forced expression of glypican-3 in hepatocellular carcinoma cells. Mol Cells. 21:224–228. 2006.PubMed/NCBI
15	Ruan J, Liu F, Chen X, et al: Inhibition of glypican-3 expression via RNA interference influences the growth and invasive ability of the MHCC97-H human hepatocellular carcinoma cell line. Int J Mol Med. 28:497–503. 2011.PubMed/NCBI
16	Kittaka N, Takemasa I, Takeda Y, et al: Molecular mapping of human hepatocellular carcinoma provides deeper biological insight from genomic data. Eur J Cancer. 44:885–897. 2008. View Article : Google Scholar : PubMed/NCBI
17	Farooq M, Hwang SY, Park MK, Kim JC, Kim MK and Sung YK: Blocking endogenous glypican-3 expression releases Hep 3B cells from G1 arrest. Mol Cells. 15:356–360. 2003.PubMed/NCBI
18	Sung YK, Hwang SY, Farooq M, Kim JC and Kim MK: Growth promotion of HepG2 hepatoma cells by antisense-mediated knockdown of glypican-3 is independent of insulin-like growth factor 2 signaling. Exp Mol Med. 35:257–262. 2003. View Article : Google Scholar : PubMed/NCBI
19	Oliver F, Christians JK, Liu X, et al: Regulatory variation at glypican-3 underlies a major growth QTL in mice. PLoS Biol. 3:e1352005. View Article : Google Scholar : PubMed/NCBI
20	Liu B, Paranjpe S, Bowen WC, et al: Investigation of the role of glypican 3 in liver regeneration and hepatocyte proliferation. Am J Pathol. 175:717–724. 2009. View Article : Google Scholar : PubMed/NCBI
21	Liu B, Bell AW, Paranjpe S, et al: Suppression of liver regeneration and hepatocyte proliferation in hepatocyte-targeted glypican 3 transgenic mice. Hepatology. 52:1060–1067. 2010. View Article : Google Scholar : PubMed/NCBI
22	Lin CW, Mars WM, Paranjpe S, et al: Hepatocyte proliferation and hepatomegaly induced by phenobarbital and 1,4-bis [2-(3,5-dichloropyridyloxy)] benzene is suppressed in hepatocyte-targeted glypican 3 transgenic mice. Hepatology. 54:620–630. 2011.PubMed/NCBI
23	Pilia G, Hughes-Benzie RM, MacKenzie A, et al: Mutations in GPC3, a glypican gene, cause the Simpson-Golabi-Behmel overgrowth syndrome. Nat Genet. 12:241–247. 1996. View Article : Google Scholar : PubMed/NCBI
24	Lapunzina P: Risk of tumorigenesis in overgrowth syndromes: a comprehensive review. Am J Med Genet C Semin Med Genet. 137C:53–71. 2005. View Article : Google Scholar : PubMed/NCBI
25	Zittermann SI, Capurro MI, Shi W and Filmus J: Soluble glypican 3 inhibits the growth of hepatocellular carcinoma in vitro and in vivo. Int J Cancer. 126:1291–1301. 2010.PubMed/NCBI
26	Feng M, Kim H, Phung Y and Ho M: Recombinant soluble glypican 3 protein inhibits the growth of hepatocellular carcinoma in vitro. Int J Cancer. 128:2246–2247. 2011. View Article : Google Scholar : PubMed/NCBI
27	Bansal R: Fibroblast growth factors and their receptors in oligodendrocyte development: implications for demyelination and remyelination. Dev Neurosci. 24:35–46. 2002. View Article : Google Scholar : PubMed/NCBI
28	Acevedo VD, Ittmann M and Spencer DM: Paths of FGFR-driven tumorigenesis. Cell Cycle. 8:580–588. 2009. View Article : Google Scholar : PubMed/NCBI
29	Pollak M: Insulin and insulin-like growth factor signalling in neoplasia. Nat Rev Cancer. 8:915–928. 2008. View Article : Google Scholar : PubMed/NCBI
30	Peters MG, Farías E, Colombo L, Filmus J, Puricelli L and Bal de Kier Joffé E: Inhibition of invasion and metastasis by glypican-3 in a syngeneic breast cancer model. Breast Cancer Res Treat. 80:221–232. 2003. View Article : Google Scholar : PubMed/NCBI
31	Lin H, Huber R, Schlessinger D and Morin PJ: Frequent silencing of the GPC3 gene in ovarian cancer cell lines. Cancer Res. 59:807–810. 1999.PubMed/NCBI
32	Kim H, Xu GL, Borczuk AC, et al: The heparan sulfate proteoglycan GPC3 is a potential lung tumor suppressor. Am J Respir Cell Mol Biol. 29:694–701. 2003. View Article : Google Scholar : PubMed/NCBI
33	Murthy SS, Shen T, De Rienzo A, et al: Expression of GPC3, an X-linked recessive overgrowth gene, is silenced in malignant mesothelioma. Oncogene. 19:410–416. 2000. View Article : Google Scholar : PubMed/NCBI
34	Xiang YY, Ladeda V and Filmus J: Glypican-3 expression is silenced in human breast cancer. Oncogene. 20:7408–7412. 2001. View Article : Google Scholar : PubMed/NCBI