Comprehensive analysis of the independent effect of twist... : Hepatology (original) (raw)

Hepatobiliary Malignancies

Comprehensive analysis of the independent effect of twist and snail in promoting metastasis of hepatocellular carcinoma#

Yang, Muh-Hwa1,2,3†; Chen, Chih-Li4†; Chau, Gar-Yang5,6; Chiou, Shih-Hwa1,7; Su, Chien-Wei1,8; Chou, Teh-Ying1,9; Peng, Wei-Li7; Wu, Jaw-Ching1,3,7*‡

1 Institutes of Clinical Medicine, National Yang-Ming University, Taipei, Taiwan

2 Division of Hematology-Oncology, Taipei Veterans General Hospital, Taipei, Taiwan

3 Genomic Medicine Research Center, Taipei Veterans General Hospital, Taipei, Taiwan

4 School of Medicine, Fu-Jen Catholic University, Taipei, Taiwan

5 Faculty of Medicine, National Yang-Ming University, Taipei, Taiwan

6 Division of General Surgery, Department of Surgery, Taipei Veterans General Hospital, Taipei, Taiwan

7 Department of Medical Research and Education, Taipei Veterans General Hospital, Taipei, Taiwan

8 Gastroenterology, Department of Medicine, Taipei Veterans General Hospital, Taipei, Taiwan

9 Department of Pathology, Taipei Veterans General Hospital, Taipei, Taiwan

* Department of Medical Research and Education, Taipei Veterans General Hospital, No. 201, Sec. 2, Shih-Pai Road, Taipei 112, Taiwan

Email:[email protected]

Received March 30, 2009; accepted July 9, 2009.

Published online 10 August 2009 in Wiley InterScience (www.interscience.wiley.com).

Grant sponsor: National Science Council; Grant Numbers: NSC93-2321-B-010-01294-2321-B-010-010 95-2321-B-010-005 98-3112-B-010-017; Grant sponsor: Ministry of Education, Aim for the Top University Plan; Grant Numbers: 96A-D-T304 97A-C-T501; Grant sponsor: Taipei Veterans General Hospital; Grant Numbers: V97ER2-016 V98ER2-017; Grant sponsor: Fu-Jen Catholic University, Taipei, Taiwan; Grant Numbers: 109531030990-3 109631030990-3.

# Potential conflict of interest: Nothing to report.

† These authors contribute equally to this study.

‡ fax: 886-2-28745074

Additional Supporting Information may be found in the online version of this article.

Abstract

The epithelial-mesenchymal transition (EMT) is critical for induction of invasiveness and metastasis of human cancers. In this study we investigated the expression profiles of the EMT markers, the relationship between EMT markers and patient/tumor/viral factors, and the interplay between major EMT regulators in human hepatocellular carcinoma (HCC). Reduced E-cadherin and nonmembranous β-catenin expression, the hallmarks of EMT, were shown in 60.2% and 51.5% of primary HCC samples, respectively. Overexpression of Snail, Twist, or Slug, the major regulators of EMT, was identified in 56.9%, 43.1%, and 51.4% of primary HCCs, respectively. Statistical analysis determined that Snail and Twist, but not Slug, are major EMT inducers in HCC: overexpression of Snail and/or Twist correlated with down-regulation of E-cadherin, nonmembranous expression of β-catenin, and a worse prognosis. In contrast, there were no such significant differences in samples that overexpressed Slug. Coexpression of Snail and Twist correlated with the worst prognosis of HCC. Hepatitis C-associated HCC was significantly correlated with Twist overexpression. HCC cell lines with increased Snail and Twist expression (e.g., Mahlavu) exhibited a greater capacity for invasiveness/metastasis than cells with low endogenous Twist/Snail expression (e.g., Huh-7). Overexpression of Snail or/and Twist in Huh-7 induced EMT and invasiveness/metastasis, whereas knockdown of Twist or Snail in Mahlavu reversed EMT and inhibited invasiveness/metastasis. Twist and Snail were independently regulated, but exerted an additive inhibitory effect to suppress E-cadherin transcription. Conclusion: Our study provides a comprehensive profile of EMT markers in HCC, and the independent and collaborative effects of Snail and Twist on HCC metastasis were confirmed through different assays. (Hepatology 2009.)