Slug expression is an independent prognostic parameter for poor survival in colorectal carcinoma patients

Abstract

Slug, a member of the Snail family of transcription factors, plays a crucial role in the regulation of epithelial-mesenchymal transition (EMT) by suppressing several epithelial markers and adhesion molecules including E-cadherin. Recently, several studies have reported Slug to be expressed in breast carcinoma, oesophageal carcinoma accompanied with shorter survival. In this study, we first investigated expression of Slug mRNA in five colorectal carcinoma cell lines by reverse transcription-polymerase chain reaction. Furthermore, we investigated Slug and E-cadherin expression by immunohistochemistry in 138 patients with colorectal carcinoma. Slug mRNA was clearly expressed in four out of five colorectal carcinoma cell lines. Positive expression of Slug and E-cadherin was observed in 37 and 58% of cases, respectively. The positive expression of Slug was significantly associated with Dukes stage and distant metastasis (P = 0.0027 and 0.0007), and the positive expression of Slug had a significant impact on patient overall survival (P < 0.0001, log-rank test). Moreover, patients with positive expression of Slug and reduced expression of E-cadherin showed the worst prognosis (P < 0.0001, log-rank test). Multivariate analysis indicated that Slug expression was an independent prognostic factor. These results suggest that positive Slug expression in colorectal carcinoma patients may become a significant parameter of poor prognosis.

Figure 1

Expression of Slug mRNA in colorectal carcinoma cell lines; DLD1, HT29, WiDr, Colo320DM, SW620. Reverse transcription-polymerase chain reaction was performed and PCR product samples were subjected to 2% agarose gel electrophoresis and visualized by staining with ethidium bromide.

Figure 2

An immunohistochemical analysis for Slug and E-cadherin in colorectal carcinoma. (A) Positive expression of Slug in the tumour cells (×200). (B) Negative expression of Slug in the tumour cells (×200). Inset, negative staining in the normal mucosa (×200). (C) Preserved E-cadherin expression is detected at the cell–cell borders and the cell membrane (×200). (D) Example of reduced expression of E-cadherin (×200).

Figure 3

(A) Overall survival of all patients (Dukes A–D) in relation to the expression of Slug (P<0.0001). (B) Overall survival of all patient's (Dukes A–D) in relation to the expression of E-cadherin (P=0.0066). (C) Overall survival of all patient's (Dukes A–D) in relation to the combination of Slug and E-cadherin expression (P<0.0001). (D) Overall survival of Dukes B, C patient's in relation to the expression of Slug (P=0.0002). (E) Overall survival of Dukes B, C patient's in relation to the expression of E-cadherin (P=0.5455). (F) Overall survival of Dukes B, C patient's in relation to the combination of Slug and E-cadherin expression (P=0.0103). Slug(+); positive expression of Slug, Slug(−); negative expression of Slug, E-cad(+); preserved E-cadherin expression, E-cad(−); reduced E-cadherin expression.