Role of Glypican-3 in the growth, migration and invasion of primary hepatocytes isolated from patients with hepatocellular carcinoma

Mauro Montalbano, Cristiana Rastellini, Joshua T. McGuire, Janika Prajapati, Ali Shirafkan, Renza Vento, Luca Cicalese

Research output: Contribution to journalArticle

5 Citations (Scopus)

Abstract

Background: Recently, Glypican-3 (GPC3) has been identified as a potential hepatocellular carcinoma (HCC) diagnostic and/or therapeutic target. GPC3 has been found to be up-regulated in HCC and to be absent in normal and cirrhotic liver. As yet, however, the molecular characteristics of GPC3 and its role in HCC cell physiology and development are still undefined. Methods: Human hepatocyte cultures were established from 10 HCC patients. Additional liver samples were obtained from 5 patients without cirrhosis and/or HCC. Soft agar colony formation, (co-)immunofluorescence and Western blot assays were used to characterize the hapatocyte cultures. The expression of GPC3 in the hepatocytes was silenced using siRNA, after which, apoptosis, scratch wound migration and transwell invasion assays were performed. Results: We found that in HCC precursor hepatocytes GPC3 is increasingly expressed in different forms and at different locations, i.e., a non-cleaved form (70 kDa) was found to be localized in the cytoplasm while a N-terminal cleaved form (N-GPC3: 40 kDa) was fond to be localized in the cytoplasm and at the extracellular side of hepatocyte membranes. In addition, we found that the non-cleaved form of GPC3 co-localizes with Furin-Convertase in the Golgi apparatus. We also found that, similar to GPC3, Furin-Convertase is expressed in HCC precursor cells, suggesting a role in GPC3 processing. Subsequent siRNA-mediated GPC3 silencing resulted in a temporary inhibition of cell proliferation, migration and ivasion, while inducing apoptosis in transformed hepatocytes. Conclusion: Our data reveal new aspects of the role of GPC3 in early hepatocyte transformation. In addition we conclude that GPC3 may serve as a new HCC immune-therapeutic target.

Original languageEnglish (US)
Pages (from-to)1-16
Number of pages16
JournalCellular Oncology
DOIs
StateAccepted/In press - Dec 4 2017

Fingerprint

Glypicans
Hepatocytes
Hepatocellular Carcinoma
Growth
Furin
Small Interfering RNA
Cytoplasm
Apoptosis
Cell Physiological Phenomena
Liver
Golgi Apparatus
Agar

Keywords

  • Cirrhotic liver
  • Furin-Convertase
  • Glypican-3 (GPC3)
  • Hepatocellular carcinoma (HCC)
  • siRNA silencing

ASJC Scopus subject areas

  • Molecular Medicine
  • Oncology
  • Cancer Research

Cite this

Role of Glypican-3 in the growth, migration and invasion of primary hepatocytes isolated from patients with hepatocellular carcinoma. / Montalbano, Mauro; Rastellini, Cristiana; McGuire, Joshua T.; Prajapati, Janika; Shirafkan, Ali; Vento, Renza; Cicalese, Luca.

In: Cellular Oncology, 04.12.2017, p. 1-16.

Research output: Contribution to journalArticle

Montalbano, Mauro ; Rastellini, Cristiana ; McGuire, Joshua T. ; Prajapati, Janika ; Shirafkan, Ali ; Vento, Renza ; Cicalese, Luca. / Role of Glypican-3 in the growth, migration and invasion of primary hepatocytes isolated from patients with hepatocellular carcinoma. In: Cellular Oncology. 2017 ; pp. 1-16.
@article{63f5034c54ed48d892477724001f72ff,
title = "Role of Glypican-3 in the growth, migration and invasion of primary hepatocytes isolated from patients with hepatocellular carcinoma",
abstract = "Background: Recently, Glypican-3 (GPC3) has been identified as a potential hepatocellular carcinoma (HCC) diagnostic and/or therapeutic target. GPC3 has been found to be up-regulated in HCC and to be absent in normal and cirrhotic liver. As yet, however, the molecular characteristics of GPC3 and its role in HCC cell physiology and development are still undefined. Methods: Human hepatocyte cultures were established from 10 HCC patients. Additional liver samples were obtained from 5 patients without cirrhosis and/or HCC. Soft agar colony formation, (co-)immunofluorescence and Western blot assays were used to characterize the hapatocyte cultures. The expression of GPC3 in the hepatocytes was silenced using siRNA, after which, apoptosis, scratch wound migration and transwell invasion assays were performed. Results: We found that in HCC precursor hepatocytes GPC3 is increasingly expressed in different forms and at different locations, i.e., a non-cleaved form (70 kDa) was found to be localized in the cytoplasm while a N-terminal cleaved form (N-GPC3: 40 kDa) was fond to be localized in the cytoplasm and at the extracellular side of hepatocyte membranes. In addition, we found that the non-cleaved form of GPC3 co-localizes with Furin-Convertase in the Golgi apparatus. We also found that, similar to GPC3, Furin-Convertase is expressed in HCC precursor cells, suggesting a role in GPC3 processing. Subsequent siRNA-mediated GPC3 silencing resulted in a temporary inhibition of cell proliferation, migration and ivasion, while inducing apoptosis in transformed hepatocytes. Conclusion: Our data reveal new aspects of the role of GPC3 in early hepatocyte transformation. In addition we conclude that GPC3 may serve as a new HCC immune-therapeutic target.",
keywords = "Cirrhotic liver, Furin-Convertase, Glypican-3 (GPC3), Hepatocellular carcinoma (HCC), siRNA silencing",
author = "Mauro Montalbano and Cristiana Rastellini and McGuire, {Joshua T.} and Janika Prajapati and Ali Shirafkan and Renza Vento and Luca Cicalese",
year = "2017",
month = "12",
day = "4",
doi = "10.1007/s13402-017-0364-2",
language = "English (US)",
pages = "1--16",
journal = "Cellular oncology (Dordrecht)",
issn = "2211-3428",
publisher = "Springer Netherlands",

}

TY - JOUR

T1 - Role of Glypican-3 in the growth, migration and invasion of primary hepatocytes isolated from patients with hepatocellular carcinoma

AU - Montalbano, Mauro

AU - Rastellini, Cristiana

AU - McGuire, Joshua T.

AU - Prajapati, Janika

AU - Shirafkan, Ali

AU - Vento, Renza

AU - Cicalese, Luca

PY - 2017/12/4

Y1 - 2017/12/4

N2 - Background: Recently, Glypican-3 (GPC3) has been identified as a potential hepatocellular carcinoma (HCC) diagnostic and/or therapeutic target. GPC3 has been found to be up-regulated in HCC and to be absent in normal and cirrhotic liver. As yet, however, the molecular characteristics of GPC3 and its role in HCC cell physiology and development are still undefined. Methods: Human hepatocyte cultures were established from 10 HCC patients. Additional liver samples were obtained from 5 patients without cirrhosis and/or HCC. Soft agar colony formation, (co-)immunofluorescence and Western blot assays were used to characterize the hapatocyte cultures. The expression of GPC3 in the hepatocytes was silenced using siRNA, after which, apoptosis, scratch wound migration and transwell invasion assays were performed. Results: We found that in HCC precursor hepatocytes GPC3 is increasingly expressed in different forms and at different locations, i.e., a non-cleaved form (70 kDa) was found to be localized in the cytoplasm while a N-terminal cleaved form (N-GPC3: 40 kDa) was fond to be localized in the cytoplasm and at the extracellular side of hepatocyte membranes. In addition, we found that the non-cleaved form of GPC3 co-localizes with Furin-Convertase in the Golgi apparatus. We also found that, similar to GPC3, Furin-Convertase is expressed in HCC precursor cells, suggesting a role in GPC3 processing. Subsequent siRNA-mediated GPC3 silencing resulted in a temporary inhibition of cell proliferation, migration and ivasion, while inducing apoptosis in transformed hepatocytes. Conclusion: Our data reveal new aspects of the role of GPC3 in early hepatocyte transformation. In addition we conclude that GPC3 may serve as a new HCC immune-therapeutic target.

AB - Background: Recently, Glypican-3 (GPC3) has been identified as a potential hepatocellular carcinoma (HCC) diagnostic and/or therapeutic target. GPC3 has been found to be up-regulated in HCC and to be absent in normal and cirrhotic liver. As yet, however, the molecular characteristics of GPC3 and its role in HCC cell physiology and development are still undefined. Methods: Human hepatocyte cultures were established from 10 HCC patients. Additional liver samples were obtained from 5 patients without cirrhosis and/or HCC. Soft agar colony formation, (co-)immunofluorescence and Western blot assays were used to characterize the hapatocyte cultures. The expression of GPC3 in the hepatocytes was silenced using siRNA, after which, apoptosis, scratch wound migration and transwell invasion assays were performed. Results: We found that in HCC precursor hepatocytes GPC3 is increasingly expressed in different forms and at different locations, i.e., a non-cleaved form (70 kDa) was found to be localized in the cytoplasm while a N-terminal cleaved form (N-GPC3: 40 kDa) was fond to be localized in the cytoplasm and at the extracellular side of hepatocyte membranes. In addition, we found that the non-cleaved form of GPC3 co-localizes with Furin-Convertase in the Golgi apparatus. We also found that, similar to GPC3, Furin-Convertase is expressed in HCC precursor cells, suggesting a role in GPC3 processing. Subsequent siRNA-mediated GPC3 silencing resulted in a temporary inhibition of cell proliferation, migration and ivasion, while inducing apoptosis in transformed hepatocytes. Conclusion: Our data reveal new aspects of the role of GPC3 in early hepatocyte transformation. In addition we conclude that GPC3 may serve as a new HCC immune-therapeutic target.

KW - Cirrhotic liver

KW - Furin-Convertase

KW - Glypican-3 (GPC3)

KW - Hepatocellular carcinoma (HCC)

KW - siRNA silencing

UR - http://www.scopus.com/inward/record.url?scp=85036579367&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=85036579367&partnerID=8YFLogxK

U2 - 10.1007/s13402-017-0364-2

DO - 10.1007/s13402-017-0364-2

M3 - Article

C2 - 29204978

AN - SCOPUS:85036579367

SP - 1

EP - 16

JO - Cellular oncology (Dordrecht)

JF - Cellular oncology (Dordrecht)

SN - 2211-3428

ER -