EBP50 inhibits pancreatic cancer cell growth and invasion by targeting the β-catenin/E-cadherin pathway

Abstract

Ezrin-radixin-moesin (ERM)-binding phosphoprotein 50 (EBP50) has previously been demonstrated to be associated with the malignant transformation of numerous types of human cancer. The aim of the present study was to investigate the effect of EBP50 overexpression on pancreatic cancer and the underlying mechanism. Reverse transcription-quantitative polymerase chain reaction was used to detect the expression of EBP50 in human pancreatic cancer tissue specimens. Furthermore, pBK-CMV-HA-EBP50 and the pBK-CMV-HA vectors were transfected into pancreatic cancer cells and the effect of EBP50 upregulation on the proliferation and invasion of the cells was investigated. In addition, the effect of EBP50 overexpression on β-catenin and E-cadherin expression was evaluated. The results revealed that overexpression of EBP50 suppressed cell growth and invasion in two human pancreatic cancer cell lines. Overexpression of EBP50 also suppressed β-catenin expression and increased E-cadherin expression. Thus, the present study demonstrated that EBP50 inhibits pancreatic cancer cell growth and invasion through targeting the β-catenin/E-cadherin pathway. The results suggest that EBP50 may function as a potential tumor suppressor and thus may serve as a potential therapeutic target.

Keywords: E-cadherin; EBP50; growth; invision; pancreatic cancer; β-catenin.

Figure 1.

(A) Immunohistochemical analysis of EBP50 in pancreatic cancer tissues. (a) No EBP50 protein expression; (b) a positive case scoring 1+; (c) a positive case scoring 2+ and (d) a positive case scoring 3+ (original magnification, x200). (B) EBP50 mRNA levels were determined by reverse transcription-quantitative polymerase chain reaction and EBP50 expression was found to be significantly decreased in pancreatic cancer tissues compared with noncancerous tissues. EBP50, ezrin-radixin-moesin-binding phosphoprotein 50.

Figure 1.

(A) Immunohistochemical analysis of EBP50 in pancreatic cancer tissues. (a) No EBP50 protein expression; (b) a positive case scoring 1+; (c) a positive case scoring 2+ and (d) a positive case scoring 3+ (original magnification, x200). (B) EBP50 mRNA levels were determined by reverse transcription-quantitative polymerase chain reaction and EBP50 expression was found to be significantly decreased in pancreatic cancer tissues compared with noncancerous tissues. EBP50, ezrin-radixin-moesin-binding phosphoprotein 50.

Figure 2.

Overexpression of EBP50 suppressed cell growth in two human pancreatic cancer cell lines as determined by cell counting kit-8 (CCK-8) assay. Absorbance values were measured at 450 nm with a microplate reader. (A) SW1990 cell and (B) PANC-1 cell groups. All experiments were performed in triplicate. Data are presented as mean ± standard deviation. * P<0.05 vs. the corresponding control group. EBP50, ezrin-radixin-moesin-binding phosphoprotein 50; HA-SW1990, SW1990 cells transfected with pBK-CMV-HA empty vector; EBP50-SW1990, SW1990 cells transfected with pBK-CMV-HV-EBP50.

Figure 3.

Anchorage-independent growth comparison of parental, pBK-CMV-HA-EBP50- and pBK-CMV-HA-transfected cells as evaluated by colony formation assay. (A-C) Representative data of SW1990 cells: (A) SW1990; (B) HA-SW1990 and; (C) EBP50-SW1990 cells. (D and E) Representative data of PANC-1 cells: (D) PANC-1; (F) HA- PANC-1 and; (F) EBP50- PANC-1 cells. Magnification, x100. EBP50, ezrin-radixin-moesin-binding phosphoprotein 50; HA-SW1990, SW1990 cells transfected with pBK-CMV-HA empty vector; EBP50-SW1990, SW1990 cells transfected with pBK-CMV-HV-EBP50.

Figure 4.

Overexpression of EBP50 inhibited the migration and invasion of pancreatic cancer cells. (A-C) Representative data of SW1990 cells: (A) SW1990; (B) HA-SW1990 and; (C) EBP50-SW1990 cells. (D and E) Representative data of PANC-1 cells: (D) PANC-1; (F) HA- PANC-1 and; (F) EBP50- PANC-1 cells. Magnification, x200. EBP50, ezrin-radixin-moesin-binding phosphoprotein 50; HA-SW1990, SW1990 cells transfected with pBK-CMV-HA empty vector; EBP50-SW1990, SW1990 cells transfected with pBK-CMV-HV-EBP50.

Figure 5.

β-catenin, and E-cadherin detected by western blotting. Representative data of: Lane 1, SW1990 cells; lane 2, HA-SW1990 cells; lane 3, EBP50-SW1990 cells; lane 4, PANC-1 cells; lane 5, HA- PANC-1 cells; and lane 6, EBP50- PANC-1 cells. EBP50, ezrin-radixin-moesin-binding phosphoprotein 50; HA-SW1990, SW1990 cells transfected with pBK-CMV-HA empty vector; EBP50-SW1990, SW1990 cells transfected with pBK-CMV-HV-EBP50.