J Pathol Transl Med.  2015 Sep;49(5):373-381. 10.4132/jptm.2015.07.09.

SALL4 Expression in Hepatocellular Carcinomas Is Associated with EpCAM-Positivity and a Poor Prognosis

Affiliations
  • 1Department of Pathology, Seoul National University Bundang Hospital, Seoul National University College of Medicine, Seoul, Korea. haeryoung.kim@snu.ac.kr
  • 2Department of Pathology, Kyungpook National University Medical Center, Kyungpook National University School of Medicine, Daegu, Korea.
  • 3Department of Surgery, Seoul National University Bundang Hospital, Seoul National University College of Medicine, Seoul, Korea.
  • 4Department of Pathology, Yonsei University College of Medicine, Seoul, Korea. young0608@yuhs.ac

Abstract

BACKGROUND
There is increasing interest in hepatocellular carcinomas (HCC) expressing "stemness"-related markers, as they have been associated with aggressive behavior and poor prognosis. In this study, we investigated the usefulness of Sal-like protein 4 (SALL4), a recently proposed candidate marker of "stemness."
METHODS
Immunohistochemical stains were performed for SALL4, K19, and epithelial cellular adhesion molecule (EpCAM) on tissue microarrays constructed from 190 surgically resected HCCs, and the results were correlated with the clinicopathological features and patient survival data.
RESULTS
Nuclear SALL4 expression was observed in 39/190 HCCs (20.5%), while K19 and EpCAM were expressed in 30 (15.9%) and 92 (48.7%) HCCs, respectively. The nuclear expression was generally weak, punctate or clumped. SALL4 expression was significantly associated with a poor overall survival compared to SALL4-negative HCCs (p = .014) compared to SALL4-negative HCCs. On multivariate analysis adjusted for tumor size, multiplicity, vascular invasion, and pathological tumor stage, SALL4 remained as a significant independent predictor of decreased overall survival (p= .004). SALL4 expression was positively correlated with EpCAM expression (p = .013) but not with K19 expression. HCCs that expressed both SALL4 and EpCAM were associated with significantly decreased overall survival, compared to those cases which were negative for both of these markers (p = .031).
CONCLUSIONS
Although SALL4 expression was not significantly correlated with other clinicopathological parameters suggestive of tumor aggressiveness, SALL4 expression was an independent predictor of poor overall survival in human HCCs, and was also positively correlated with EpCAM expression.

Keyword

Carcinoma, hepatocellular; SALL4; Immunohistochemistry; Prognosis

MeSH Terms

Carcinoma, Hepatocellular*
Coloring Agents
Humans
Immunohistochemistry
Multivariate Analysis
Prognosis*
Coloring Agents

Figure

  • Fig. 1. Immunohistochemical stain results. Nuclear SALL4 staining is seen in a case of HCC (A), which is weaker and clumped (inset) compared to the strong nuclear staining seen in testicular seminoma (B). Clumped nuclear SALL4 expression is seen in another case of HCC with fibrous stroma (C). The non-neoplastic liver is negative for SALL4, except for occasional faintly positive cells in the ductular reactions (D, arrows). Nuclear SALL4 staining (E) is seen in an HCC with EpCAM expression (F). SALL4, Sal-like protein 4; HCC, hepatocellular carcinoma; EpCAM, epithelial cellular adhesion molecule.

  • Fig. 2. Venn diagram summarizing the relationships between the expression of SALL4 (pink), K19 (blue), and EpCAM (green) in HCCs. While SALL4 expression is frequently co-expressed with EpCAM (26/39, 66.7%), K19 expression was only seen in 6 of SALL4-positive HCCs (15.4%). SALL4, Sal-like protein 4; EpCAM, epithelial cellular adhesion molecule; HCC, hepatocellular carcinoma. aTotal n=189 with SALL4, EpCAM, and K19 data.

  • Fig. 3. Kaplan-Meier survival curves demonstrating decreased overall survival (A) and disease-free survival (B) in HCCs with SALL4 expression, and decreased overall survival (C) and disease-free survival (D) in HCCs with K19 expression. HCC, hepatocellular carcinoma; SALL4, Sal-like protein 4.

  • Fig. 4. Kaplan-Meier survival curves demonstrating the differences in overall survival in HCCs after combining EpCAM and SALL4 expression status. HCC, hepatocellular carcinoma; EpCAM, epithelial cellular adhesion molecule; SALL4, Sal-like protein 4.


Cited by  1 articles

A New Cell Block Method for Multiple Immunohistochemical Analysis of Circulating Tumor Cells in Patients with Liver Cancer
Soo Jeong Nam, Hyun Yang Yeo, Hee Jin Chang, Bo Hyun Kim, Eun Kyung Hong, Joong-Won Park
Cancer Res Treat. 2016;48(4):1229-1242.    doi: 10.4143/crt.2015.500.


Reference

1. Lee JS, Heo J, Libbrecht L, et al. A novel prognostic subtype of human hepatocellular carcinoma derived from hepatic progenitor cells. Nat Med. 2006; 12:410–6.
Article
2. Nault JC, De Reynies A, Villanueva A, et al. A hepatocellular carcinoma 5-gene score associated with survival of patients after liver resection. Gastroenterology. 2013; 145:176–87.
Article
3. Woo HG, Lee JH, Yoon JH, et al. Identification of a cholangiocarcinoma-like gene expression trait in hepatocellular carcinoma. Cancer Res. 2010; 70:3034–41.
Article
4. Yamashita T, Forgues M, Wang W, et al. EpCAM and alpha-fetoprotein expression defines novel prognostic subtypes of hepatocellular carcinoma. Cancer Res. 2008; 68:1451–61.
5. Kim H, Choi GH, Na DC, et al. Human hepatocellular carcinomas with “Stemness”-related marker expression: keratin 19 expression and a poor prognosis. Hepatology. 2011; 54:1707–17.
Article
6. Kim H, Park YN. Hepatocellular carcinomas expressing ‘stemness’-related markers: clinicopathological characteristics. Dig Dis. 2014; 32:778–85.
Article
7. Govaere O, Komuta M, Berkers J, et al. Keratin 19: a key role player in the invasion of human hepatocellular carcinomas. Gut. 2014; 63:674–85.
Article
8. Kim H, Yoo JE, Cho JY, et al. Telomere length, TERT and shelterin complex proteins in hepatocellular carcinomas expressing “stemness”-related markers. J Hepatol. 2013; 59:746–52.
Article
9. Oikawa T, Kamiya A, Zeniya M, et al. Sal-like protein 4 (SALL4), a stem cell biomarker in liver cancers. Hepatology. 2013; 57:1469–83.
Article
10. Yong KJ, Gao C, Lim JS, et al. Oncofetal gene SALL4 in aggressive hepatocellular carcinoma. N Engl J Med. 2013; 368:2266–76.
Article
11. Zeng SS, Yamashita T, Kondo M, et al. The transcription factor SALL4 regulates stemness of EpCAM-positive hepatocellular carcinoma. J Hepatol. 2014; 60:127–34.
Article
12. Zhang J, Tam WL, Tong GQ, et al. SALL4 modulates embryonic stem cell pluripotency and early embryonic development by the transcriptional regulation of Pou5f1. Nat Cell Biol. 2006; 8:1114–23.
Article
13. Wu Q, Chen X, Zhang J, et al. SALL4 interacts with Nanog and cooccupies Nanog genomic sites in embryonic stem cells. J Biol Chem. 2006; 281:24090–4.
Article
14. Kohlhase J, Heinrich M, Schubert L, et al. Okihiro syndrome is caused by SALL4 mutations. Hum Mol Genet. 2002; 11:2979–87.
15. Ikeda H, Sato Y, Yoneda N, et al. α-Fetoprotein-producing gastric carcinoma and combined hepatocellular and cholangiocarcinoma show similar morphology but different histogenesis with respect to SALL4 expression. Hum Pathol. 2012; 43:1955–63.
Article
16. Ushiku T, Shinozaki A, Shibahara J, et al. SALL4 represents fetal gut differentiation of gastric cancer, and is diagnostically useful in distinguishing hepatoid gastric carcinoma from hepatocellular carcinoma. Am J Surg Pathol. 2010; 34:533–40.
Article
17. Ma Y, Cui W, Yang J, et al. SALL4, a novel oncogene, is constitutively expressed in human acute myeloid leukemia (AML) and induces AML in transgenic mice. Blood. 2006; 108:2726–35.
Article
18. Cao D, Humphrey PA, Allan RW. SALL4 is a novel sensitive and specific marker for metastatic germ cell tumors, with particular utility in detection of metastatic yolk sac tumors. Cancer. 2009; 115:2640–51.
Article
19. Oikawa T, Kamiya A, Kakinuma S, et al. SALL4 regulates cell fate decision in fetal hepatic stem/progenitor cells. Gastroenterology. 2009; 136:1000–11.
Article
20. Liu TC, Vachharajani N, Chapman WC, Brunt EM. SALL4 immunoreactivity predicts prognosis in Western hepatocellular carcinoma patients but is a rare event: a study of 236 cases. Am J Surg Pathol. 2014; 38:966–72.
21. Gonzalez-Roibon N, Katz B, Chaux A, et al. Immunohistochemical expression of SALL4 in hepatocellular carcinoma, a potential pitfall in the differential diagnosis of yolk sac tumors. Hum Pathol. 2013; 44:1293–9.
Article
Full Text Links
  • JPTM
Actions
Cited
CITED
export Copy
Close
Share
  • Twitter
  • Facebook
Similar articles
Copyright © 2024 by Korean Association of Medical Journal Editors. All rights reserved.     E-mail: koreamed@kamje.or.kr