Oncogene GAEC1 regulates CAPN10 expression which predicts survival in esophageal squamous cell carcinoma

Figure 1 Expression level of GAEC1 in KYSE150 cells. A: Multiplex reverse transcription-polymerase chain reaction (RT-PCR) analysis showed the overexpression of GAEC1 in KYSE150 compared with the non-tumor esophageal epithelial cell line NE1; B: Multiplex RT-PCR analysis demonstrated the down-regulation of GAEC1 expression in KYSE150 cells transfected with pSilencer P3-4 vector compared with the parental cells and those transfected with pSilencer control vector. The amount of RNA in each lane was normalized with the amplification of β-actin. M: 100 bp ladder marker; H₂O: Water control.

Figure 2 MTS cell proliferation assays for esophageal squamous cell carcinoma cell line KYSE150. Cells were transfected with pSilencer vector cloned with the P3-4 sequence (pSilencer P3-4) or control vector (pSilencer-ve). MTS assays were then performed every 24 h for 3 d on each type of transfected cells and the parental cells. The respective MTS activities on each day were compared with the corresponding activities of day 1. Representative data from 3 independent experiments are shown.

Figure 3 Flow cytometry analyses for KYSE150 cells. KYSE150 transfected with pSilencer cloned with P3-4 sequence demonstrated an increased apoptotic population by approximately 50% (C) compared with the parental cells (A) and cells transfected with pSilencer-ve control vector (B).

Figure 4 Immunohistochemical staining of CAPN10. A: In normal esophageal epithelial tissue showing weak CAPN10 staining; B: esophageal squamous cell carcinoma (ESCC) tissue showing strong CAPN10 staining; C: ESCC tissue showing weak CAPN10 staining in tumor. CAPN10 was mainly localized in the cytoplasm of the cancer cells (original magnification, ×100).

Figure 5 Overall 5-year survival rates as determined by the expression level of CAPN10 in esophageal squamous cell carcinoma patients. Low expression group of CAPN10 in ESCC patients showed a significantly lower 5-year survival rate than those of high expression group.