Yonsei Med J.  2018 Dec;59(10):1159-1165. 10.3349/ymj.2018.59.10.1159.

PLCE1 Promotes the Invasion and Migration of Esophageal Cancer Cells by Up-Regulating the PKCα/NF-κB Pathway

• Li Y
• Luan C

• KMID: 2426329
• DOI: http://doi.org/10.3349/ymj.2018.59.10.1159

Abstract

PURPOSE
To investigate the effect and mechanism of phospholipase C epsilon gene 1 (PLCE1) expression on esophageal cancer cell lines.
MATERIALS AND METHODS
The esophageal carcinoma cell lines Eca109 and EC9706 and normal esophageal epithelial cell line HEEC were cultured. The expression of PLCE1, protein kinase C alpha (PKCα), and nuclear factor kappa B (NF-κB) p50/p65 homodimer in cells were comparatively analyzed. The esophageal cancer cells were divided into si-PLCE1, control siRNA (scramble), and mock groups that were transfected with specific siRNA for PLCE1, control siRNA, and blank controls, respectively. Expression of PLCE1, PKCα, p50, and p65 was detected by Western blotting. Transwell assay was used to detect migration and invasion of Eca109 and EC9706 cells.
RESULTS
Compared with HEEC, the expression of PLCE1, PKCα, p50, and p65 was increased in Eca109 and EC9706 cells. The expression of PLCE1 was positively correlated with the expression of PKCα and p50 (PKCα: r=0.6328, p=0.032; p50: r=0.6754, p=0.041). PKCα expression had a positive correlation with the expression of p50 and p65 (p50: r=0.9127, p=0.000; p65: r=0.9256, p=0.000). Down-regulation of PLCE1 significantly decreased the expression of PKCα and NF-κB-related proteins (p65: p=0.002, p=0.004; p50: p=0.005, p=0.009) and inhibited the migration and invasion of Eca109 and EC9706 cells.
CONCLUSION
PLCE1 activated NF-κB signaling by up-regulating PKCα, which could promote invasion and migration of esophageal cancer cells.

Keyword

Esophageal cancer; PLCE1; NF-κB signaling pathway; invasion; migration; PKCα

MeSH Terms

Blotting, Western
Cell Line
Down-Regulation
Epithelial Cells
Esophageal Neoplasms*
NF-kappa B
Protein Kinase C-alpha
RNA, Small Interfering
Type C Phospholipases
NF-kappa B
Protein Kinase C-alpha
RNA, Small Interfering
Type C Phospholipases

Figure

• 

  Fig. 1 Expression of PLCE1 and PKCα in esophageal cancer cells. (A) qRT-PCR was used to detect the expression of PLCE1 mRNA in esophageal cancer cells. (B) The expression of PLCE1 was detected by Western blotting. (C) The expression of PKCα mRNA. (D) The expression of PKCα protein and its statistical analysis map. Compared with HEEC, *p<0.01.

• 

  Fig. 2 Expression of NF-κB signaling pathway-related proteins. (A) Western blotting was used to detect the expression of p50 and p65. (B) Statistical analysis of P50 and p65 expression. Compared with HEEC, *p<0.01.

• 

  Fig. 3 Correlation analysis of PLCE1, PKCα, and p50/p65 protein expression in esophageal cancer cells. (A) Relationships among PLCE1, PKCα, and p50/p65. (B) Relationship between PKCα and p50/p65.

• 

  Fig. 4 Effect of PLCE1 on PKCα and p50/p65 expression in esophageal cancer cells. (A) After transfection with PLCE1 siRNA, the expression of PLCE1, PKCα, and p50/p65 was detected by Western blotting. (B) Statistical analysis map of PLCE1, PKCα, and p50/p65. Compared with the mock group or scramble group, *p<0.05, †p<0.01.

• 

  Fig. 5 Migration ability of Eca109 and EC9706 cells. (A) Transwell assay was used to detect the migration of esophageal cancer cells (×400). (B) Statistical map of cell migration. Compared with the mock group or scramble group, *p<0.01.

• 

  Fig. 6 Invasion ability of Eca109 and EC9706 cells. (A) Transwell assay was used to detect the invasion of esophageal cancer cells (×400). (B) Statistical map of cell invasion. Compared with the mock group or scramble group, *p<0.01.

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