J Pathol Transl Med.  2016 Sep;50(5):327-336. 10.4132/jptm.2016.06.22.

Clinicopathologic Correlations of E-cadherin and Prrx-1 Expression Loss in Hepatocellular Carcinoma

• Yi K
• Kim H
• Chung Y
• Ahn H
• Sim J
• Wi YC
• Pyo JY
• Song YS
• Paik SS
• Oh YH

• KMID: 2353594
• DOI: http://doi.org/10.4132/jptm.2016.06.22

Abstract

BACKGROUND
Developing predictive markers for hepatocellular carcinoma (HCC) is important, because many patients experience recurrence and metastasis. Epithelial to mesenchymal transition (EMT) is a developmental process that plays an important role during embryogenesis and also during cancer metastasis. Paired-related homeobox protein 1 (Prrx-1) is an EMT inducer that has recently been introduced, and its prognostic significance in HCC is largely unknown.
METHODS
Tissue microarray was constructed using surgically resected primary HCCs from 244 cases. Immunohistochemical staining of E-cadherin and Prrx-1 was performed. The correlation between E-cadherin loss and Prrx-1 expression, as well as other clinicopathologic factors, was evaluated.
RESULTS
E-cadherin expression was decreased in 96 cases (39.4%). Loss of E-cadherin correlated with a higher recurrence rate (p < .001) but was not correlated with patient's survival. Thirty-two cases (13.3%) showed at least focal nuclear Prrx-1 immunoreactivity while all non-neoplastic livers (n = 22) were negative. Prrx-1 expression was not associated with E-cadherin loss, survival or recurrence rates, pathologic factors, or the Ki-67 labeling index. Twenty tumors that were positive for E-cadherin and Prrx-1 had significantly higher nuclear grades than the rest of the cohort (p = .037). In Cox proportional hazard models, E-cadherin loss and large vessel invasion were independent prognostic factors for shorter disease-free survival. Cirrhosis and high Ki-67 index (> 40%) were independent prognostic factors for shorter overall survival.
CONCLUSIONS
Prrx-1 was expressed in small portions of HCCs but not in normal livers. Additional studies with a large number of Prrx-1-positive cases are required to confirm the results of this study.

Keyword

Liver; Neoplasms; Epithelial-mesenchymal transition; Prrx-1 protein

MeSH Terms

Cadherins*
Carcinoma, Hepatocellular*
Cohort Studies
Disease-Free Survival
Embryonic Development
Epithelial-Mesenchymal Transition
Female
Fibrosis
Genes, Homeobox
Humans
Liver
Neoplasm Metastasis
Pregnancy
Proportional Hazards Models
Recurrence
Cadherins

Figure

• 

  Fig. 1. Immunohistochemical staining of E-cadherin and paired-related homeobox protein 1 (Prrx-1) in hepatocellular carcinomas. Representative photos of immunohistochemical staining in hepatocellular carcinomas. (A) E-cadherin decreased. (B) E-cadherin maintained. (C) Prrx-1 negative. (D) Prrx-1 positive. (E) Prrx-1 negative, but cytoplasmic positive.

• 

  Fig. 2. Kaplan-Meier curves for overall survival (A, C, E) and disease-free survival (B, D, F) in hepatocellular carcinomas according to American Joint Committee on Cancer stage, E-cadherin and paired-related homeobox protein 1 (Prrx-1) expression (log-rank test).

• 

  Fig. 3. Cox proportional hazard regression model for disease-free survival. CI, confidence interval; AJCC, American Joint Committee on Cancer; Prrx-1, paired-related homeobox protein 1. a Cirrhosis vs chronic hepatitis.

• 

  Fig. 4. Cox proportional hazard regression model for overall survival. CI, confidence interval; AJCC, American Joint Committee on Cancer; Prrx-1, paired-related homeobox protein 1. a Cirrhosis vs chronic hepatitis.

Reference

1. Jung KW, Won YJ, Kong HJ, Oh CM, Seo HG, Lee JS. Cancer statistics in Korea: incidence, mortality, survival and prevalence in 2010. Cancer Res Treat. 2013; 45:1–14.

2. Tung-Ping Poon R, Fan ST, Wong J. Risk factors, prevention, and management of postoperative recurrence after resection of hepatocellular carcinoma. Ann Surg. 2000; 232:10–24.

3. Thiery JP, Acloque H, Huang RY, Nieto MA. Epithelial-mesenchymal transitions in development and disease. Cell. 2009; 139:871–90.

4. Hardin H, Guo Z, Shan W, et al. The roles of the epithelial-mesenchymal transition marker PRRX1 and miR-146b-5p in papillary thyroid carcinoma progression. Am J Pathol. 2014; 184:2342–54.

5. Ocana OH, Corcoles R, Fabra A, et al. Metastatic colonization requires the repression of the epithelial-mesenchymal transition inducer Prrx1. Cancer Cell. 2012; 22:709–24.

6. Reichert M, Takano S, von Burstin J, et al. The Prrx1 homeodomain transcription factor plays a central role in pancreatic regeneration and carcinogenesis. Genes Dev. 2013; 27:288–300.

7. Takahashi Y, Sawada G, Kurashige J, et al. Paired related homoeobox 1, a new EMT inducer, is involved in metastasis and poor prognosis in colorectal cancer. Br J Cancer. 2013; 109:307–11.

8. Hirata H, Sugimachi K, Takahashi Y, et al. Downregulation of PRRX1 confers cancer stem cell-like properties and predicts poor prognosis in hepatocellular carcinoma. Ann Surg Oncol. 2015; 22 Suppl 3:S1402–9.

9. Woo HY, Min AL, Choi JY, Bae SH, Yoon SK, Jung CK. Clinicopathologic significance of the expression of Snail in hepatocellular carcinoma. Korean J Hepatol. 2011; 17:12–8.

10. Schuffler PJ, Fuchs TJ, Ong CS, Wild PJ, Rupp NJ, Buhmann JM. TMARKER: A free software toolkit for histopathological cell counting and staining estimation. J Pathol Inform. 2013; 4(Suppl):S2.

11. Nakanishi K, Sakamoto M, Yamasaki S, Todo S, Hirohashi S. Akt phosphorylation is a risk factor for early disease recurrence and poor prognosis in hepatocellular carcinoma. Cancer. 2005; 103:307–12.

12. Ito Y, Matsuura N, Sakon M, et al. Both cell proliferation and apoptosis significantly predict shortened disease-free survival in hepatocellular carcinoma. Br J Cancer. 1999; 81:747–51.

13. Prall F. Tumour budding in colorectal carcinoma. Histopathology. 2007; 50:151–62.

14. Peinado H, Olmeda D, Cano A. Snail, Zeb and bHLH factors in tumour progression: an alliance against the epithelial phenotype? Nat Rev Cancer. 2007; 7:415–28.

15. Cho SB, Lee KH, Lee JH, et al. Expression of E- and N-cadherin and clinicopathology in hepatocellular carcinoma. Pathol Int. 2008; 58:635–42.

16. Zhang L, Huang G, Li X, et al. Hypoxia induces epithelial-mesenchymal transition via activation of SNAI1 by hypoxia-inducible factor -1alpha in hepatocellular carcinoma. BMC Cancer. 2013; 13:108.

17. Kwon GY, Yoo BC, Koh KC, Cho JW, Park WS, Park CK. Promoter methylation of E-cadherin in hepatocellular carcinomas and dysplastic nodules. J Korean Med Sci. 2005; 20:242–7.

18. Chen J, Zhao J, Ma R, Lin H, Liang X, Cai X. Prognostic significance of E-cadherin expression in hepatocellular carcinoma: a meta-analysis. PLoS One. 2014; 9:e103952.

19. Aoki S, Shimamura T, Shibata T, et al. Prognostic significance of dysadherin expression in advanced colorectal carcinoma. Br J Cancer. 2003; 88:726–32.

20. Asgeirsson KS, Jonasson JG, Tryggvadóttir L, et al. Altered expression of E-cadherin in breast cancer. patterns, mechanisms and clinical significance. Eur J Cancer. 2000; 36:1098–106.

21. Zhao XL, Sun T, Che N, et al. Promotion of hepatocellular carcinoma metastasis through matrix metalloproteinase activation by epithelial-mesenchymal transition regulator Twist1. J Cell Mol Med. 2011; 15:691–700.

22. Tannapfel A, Geissler F, Köckerling F, Katalinic A, Hauss J, Wittekind C. Apoptosis and proliferation in relation to histopathological variables and prognosis in hepatocellular carcinoma. J Pathol. 1999; 187:439–45.

23. Tsai JH, Donaher JL, Murphy DA, Chau S, Yang J. Spatiotemporal regulation of epithelial-mesenchymal transition is essential for squamous cell carcinoma metastasis. Cancer Cell. 2012; 22:725–36.

24. Ferrell JE Jr, Machleder EM. The biochemical basis of an all-or-none cell fate switch in Xenopus oocytes. Science. 1998; 280:895–8.

25. Batchelor E, Loewer A, Lahav G. The ups and downs of p53: understanding protein dynamics in single cells. Nat Rev Cancer. 2009; 9:371–7.