Elevated GPC3 level promotes cell proliferation in liver cancer

Wang,Shanshan; Chen,Ning; Chen,Yuhan; Sun,Lin; Li,Li; Liu,Hui

doi:10.3892/ol.2018.8754

Elevated GPC3 level promotes cell proliferation in liver cancer

Authors:
- Shanshan Wang
- Ning Chen
- Yuhan Chen
- Lin Sun
- Li Li
- Hui Liu
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Affiliations: Beijing You'An Hospital Affiliated to Capital Medical University, Beijing Institute of Hepatology, Beijing 100069, P.R. China, Department of Gastrointestinal and Hepatology, Beijing You'An Hospital Affiliated to Capital Medical University, Beijing 100069, P.R. China, Department of Pathology, Beijing You'An Hospital Affiliated to Capital Medical University, Beijing 100069, P.R. China
Published online on: May 21, 2018 https://doi.org/10.3892/ol.2018.8754
Pages: 970-976
Copyright: © Wang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

The aim of the present study was to investigate the biological role of glypican 3 (GPC3), and to identify its mechanism and clinical significance in the carcinogenesis of liver cancer. A total of 114 patients with liver cancer were involved. Their clinical data, hematoxylin and eosin‑stained and Antigen Ki‑67 protein (Ki‑67) and GPC3 immunohistochemically‑stained liver cancer tissue sections were analyzed to evaluate the correlation between the liver cancer proliferation, differentiation and GPC3 expression. Fluorescence microscopy, western blotting, MTT and reverse transcription quantitative polymerase chain reaction (RT‑qPCR) assays were performed in HepG2 and HLE cell lines to investigate the potential mechanisms of action. Among the 114 patients with liver cancer enrolled in the present study, 12 exhibited well‑differentiated liver cancer, of which 6 (50%) were positive for GPC3. A total of 30 cases exhibited poorly differentiated liver cancer; 26 (87%) of these expressed GPC3 and 11 cases (37%) demonstrated strong positive expression levels. The other 72 liver cancer cases were moderately differentiated; 75% (54/72) of these expressed GPC3 and 12.5% (9/72) exhibited strong positive expression levels. There was a significant association between the levels of GPC3 expression and liver cancer differentiation (χ2=16.306, P=0.008). Ki‑67 staining as the criteria of the liver cancer cell proliferation index also indicated a cross correlation between liver cancer proliferation and GPC3 levels. Among the 39 liver cancer samples with a cell proliferation index <5%, only 2.6% (1/39) exhibited strong positive GPC3 staining, but of the 16 cases with a high cell proliferation index >50%, 6 exhibited strong GPC3 staining (37.5%). The difference of cell proliferation indexes between cancer cells were well, moderate and poorly differentiated, and was markedly significant (χ2=26.334, P=0.002), and suggested that liver cancer cell proliferation was positively correlated with GPC3 expression (r=0.316, P=0.001). Consistently, in vitro analysis indicated that GPC3 promoted HepG2 and HLE cell growth, which was more apparent in HepG2 cells. The RT‑qPCR results indicated that GPC3 promoted proliferation through the Hedgehog (Hh) pathway in HepG2 cells, but not in HLE cells. In the present study, it was demonstrated that patients with liver cancer with higher GPC3 levels exhibited poorer differentiation and higher proliferation levels. In vitro GPC3 may promote liver cancer cell lines proliferation through the Hh pathway.

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1	Boily G, Ouellet S, Langlois S, Larivière M, Drouin R and Sinnett D: In vivo footprinting analysis of the Glypican 3 (GPC3) promoter region in neuroblastoma cells. Biochim Biophys Acta. 1769:182–193. 2007. View Article : Google Scholar : PubMed/NCBI
2	Filmus J and Selleck SB: Glypicans: Proteoglycans with a surprise. J Clin Invest. 108:497–501. 2001. View Article : Google Scholar : PubMed/NCBI
3	Filmus J, Capurro M and Rast J: Glypicans. GENOME BIOL 2008 2008-01-20. 9:224
4	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
5	Filmus J and Capurro M: The role of glypican-3 in the regulation of body size and cancer. Cell Cycle. 7:2787–2790. 2008. View Article : Google Scholar : PubMed/NCBI
6	Song HH and Filmus J: The role of glypicans in mammalian development. Biochim Biophys Acta. 1573:241–246. 2002. View Article : Google Scholar : PubMed/NCBI
7	Pilia G, Hughes-Benzie RM, MacKenzie A, Baybayan P, Chen EY, Huber R, Neri G, Cao A, Forabosco A and Schlessinger D: 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
8	Li M, Shuman C, Fei YL, Cutiongco E, Bender HA, Stevens C, Wilkins-Haug L, Day-Salvatore D, Yong SL, Geraghty MT, et al: GPC3 mutation analysis in a spectrum of patients with overgrowth expands the phenotype of Simpson-Golabi-Behmel syndrome. Am J Med Genet. 102:161–168. 2001. View Article : Google Scholar : PubMed/NCBI
9	Nakatsura T, Kageshita T, Ito S, Wakamatsu K, Monji M, Ikuta Y, Senju S, Ono T and Nishimura Y: Identification of glypican-3 as a novel tumor marker for melanoma. Clin Cancer Res. 10:6612–6621. 2004. View Article : Google Scholar : PubMed/NCBI
10	Maeda D, Ota S, Takazawa Y, Aburatani H, Nakagawa S, Yano T, Taketani Y, Kodama T and Fukayama M: Glypican-3 expression in clear cell adenocarcinoma of the ovary. Mod Pathol. 22:824–832. 2009. View Article : Google Scholar : PubMed/NCBI
11	Zynger DL, Dimov ND, Luan C, Teh BT and Yang XJ: Glypican 3: A novel marker in testicular germ cell tumors. Am J Surg Pathol. 30:1570–1575. 2006. View Article : Google Scholar : PubMed/NCBI
12	Schmilovitz-Weiss H, Tobar A, Halpern M, Levy I, Shabtai E and Ben-Ari Z: Tissue expression of squamous cellular carcinoma antigen and Ki67 in hepatocellular carcinoma-correlation with prognosis: a historical prospective study. Diagn Pathol. 6:1212001. View Article : Google Scholar
13	Torre LA, Bray F, Siegel RL, Ferlay J, Lortet-Tieulent J and Jemal A: Global cancer statistics, 2012. CA Cancer J Clin. 65:87–108. 2015. View Article : Google Scholar : PubMed/NCBI
14	Gao JD, Shao YF, Xu Y, Ming LH, Wu ZY, Liu GT, Wang XH, Gao WH, Sun YT, Feng XL, et al: Tight association of hepatocellular carcinoma with HBV infection in North China. Hepatobiliary Pancreat Dis Int. 4:46–49. 2005.PubMed/NCBI
15	Wang HL, Anatelli F, Zhai QJ, Adley B, Chuang ST and Yang XJ: Glypican-3 as a useful diagnostic marker that distinguishes hepatocellular carcinoma from benign hepatocellular mass lesions. Arch Pathol Lab Med. 132:1723–1728. 2008.PubMed/NCBI
16	Bosman FT, Carneiro F, Hruban RH and Theise ND: WHO classification of tumours of the digestive system. 4th edition. Int Agen Res Cancer Lyon. 1089:2010.
17	Shirakawa H, Kuronuma T, Nishimura Y, Hasebe T, Nakano M, Gotohda N, Takahashi S, Nakagohri T, Konishi M, Kobayashi N, et al: Glypican-3 is a useful diagnostic marker for a component of hepatocellular carcinoma in human liver cancer. Int J Oncol. 34:649–656. 2009.PubMed/NCBI
18	López-Terrada D, Cheung SW, Finegold MJ and Knowles BB: Hep G2 is a hepatoblastoma-derived cell line. Hum Pathol. 40:1512–1525. 2009. View Article : Google Scholar
19	Wang SS, Chen YH, Ning C, Wang LJ, Chen DX, Weng HL, Dooley S and Ding HG: Hydrogen sulfide promotes autophagy of hepatocellular carcinoma cells through the PI3K/Akt/mTOR signaling pathway. Cell Death Dis. 8:e26882017. View Article : Google Scholar : PubMed/NCBI
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI
21	Kawano Y, Sasaki A, Kai S, Endo Y, Iwaki K, Uchida H, Shibata K, Ohta M and Kitano S: Short- and long-term outcomes after hepatic resection for hepatocellular carcinoma with concomitant esophageal varices in patients with cirrhosis. Ann Surg Oncol. 15:1670–1676. 2008. View Article : Google Scholar : PubMed/NCBI
22	Liu H, Li P, Zhai Y, Qu CF, Zhang LJ, Tan YF, Li N and Ding HG: 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
23	Iglesias BV, Centeno G, Pascuccelli H, Ward F, Peters MG, Filmus J, Puricelli L and de Kier Joffé EB: Expression pattern of glypican-3 (GPC3) during human embryonic and fetal development. Histol Histopathol. 23:1333–1340. 2008.PubMed/NCBI
24	Jian R, Zheng DY and Luo RC: The expression of GPC3 in human malignant cancers and its potential clinical application. Tumor. 31:863–866. 2011.
25	Suzuki M, Sugimoto K, Tanaka J, Tameda M, Inagaki Y, Kusagawa S, Nojiri K, Beppu T, Yoneda K, Yamamoto N, et al: Up-regulation of glypican-3 in human hepatocellular carcinoma. Anticancer Res. 30:5055–5061. 2010.PubMed/NCBI
26	Lin Q, Xiong LW, Pan XF, Gen JF, Bao GL, Sha HF, Feng JX, Ji CY and Chen M: Expression of GPC3 protein and its significance in lung squamous cell carcinoma. Med Oncol. 29:663–669. 2012. View Article : Google Scholar : PubMed/NCBI
27	An SJ, Huang YS, Chen ZH, Su J, Yang Y, Chen JG, Yan HH, Lin QX, Yang JJ, Yang XN, et al: Posttreatment plasma VEGF levels may be associated with the overall survival of patients with advanced non-small cell lung cancer treated with bevacizumab plus chemotherapy. Med Oncol. 29:627–632. 2012. View Article : Google Scholar : PubMed/NCBI
28	Mounajjed T, Zhang L and Wu TT: Glypican-3 expression in gastrointestinal and pancreatic epithelial neoplasms. Hum Pathol. 44:542–550. 2013. View Article : Google Scholar : PubMed/NCBI
29	Valsechi MC, Oliveira AB, Conceição AL, Stuqui B, Candido NM, Provazzi PJ, de Araújo LF, Silva WA Jr, Calmon Mde F and Rahal P: GPC3 reduces cell proliferation in renal carcinoma cell lines. BMC Cancer. 14:6312014. View Article : Google Scholar : PubMed/NCBI
30	Stigliano I, Puricelli L, Filmus J, Sogayar MC, Bal de Kier Joffé E and Peters MG: Glypican-3 regulates migration, adhesion and actin cytoskeleton organization in mammary tumor cells through Wnt signaling modulation. Breast Cancer Res Treat. 114:251–262. 2009. View Article : Google Scholar : PubMed/NCBI
31	Gao W and Ho M: The role of glypican-3 in regulating Wnt in hepatocellular carcinomas. Cancer Rep. 1:14–19. 2011.PubMed/NCBI
32	Pan Z, Chen C, Long H, Lei C, Tang G, Li L, Feng J and Chen F: Overexpression of GPC3 inhibits hepatocellular carcinoma cell proliferation and invasion through induction of apoptosis. Mol Med Rep. 7:969–974. 2013. View Article : Google Scholar : PubMed/NCBI
33	Sakurai M, Shibata K, Umezu T, Kajiyama H, Yamamoto E, Ino K, Nawa A and Kikkawa F: 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
34	Miao HL, Pan ZJ, Lei CJ, Wen JY, Li MY, Liu ZK, Qiu ZD, Lin MZ, Chen NP and Chen M: Knockdown of GPC3 inhibits the proliferation of Huh7 hepatocellular carcinoma cells through down-regulation of YAP. J Cell Biochem. 114:625–631. 2013. View Article : Google Scholar : PubMed/NCBI