Figure 1.
Western blot analysis and quantification of 14-3-3σ protein levels in different tissue types and esophageal epithelial cell lines.
(A) Representative Western blot of 14-3-3σ protein levels and quantification in 20 samples of normal esophageal epithelia (NEE), 17 samples of low grade intraepithelial neoplasia (LGIN) and 13 samples of high grade intraepithelial neoplasia (HGIN). (B) Representative Western blot of 14-3-3σ protein levels and quantification in 80 paired samples of ESCC (T) and adjacent normal esophageal epithelia (N) with various clinical stages (20 ESCC samples for each stage). (C) Quantification of 14-3-3σ protein levels in the continuum of ESCC progression. (D) 14-3-3σ protein levels in an immortalized primary esophageal epithelial cell line NEC, three ESCC cell lines EC1, EC109 and EC9706, one paclitaxel-resistant cell line EC9706/PTX and one cisplatin-resistant cell line EC9706/CDDP. β-actin was used as a control for equal loading.
Figure 2.
Representative immunohistochemical staining of 14-3-3σ.
Normal esophageal epithelia (A), low grade intraepithelial neoplasia (B), high grade intraepithelial neoplasia (C), well-differentiated (D), moderately-differentiated (E) and poorly-differentiated ESCC (F).
Table 1.
14-3-3σ protein expression during cancer progression by IHC analysis.
Table 2.
Association between 14-3-3σ expression and clinicopathological parameters of ESCC.
Figure 3.
Kaplan-Meier survival curves of ESCC patients.
(A) The 5-year overall survival rate of 168 ESCC patients was 30.26%. (B) The 5-year overall survival rates in ESCC patients with low (n = 124) and high 14-3-3σ protein (n = 44) were 25.62% and 43.47%, respectively, with a significant difference (P = 0.004). (C) The 5-year overall survival rates were 64.66%, 33.35% in ESCC patients with high (n = 24) and low expression (n = 73) of 14-3-3σ, respectively in early stage (I–II) ESCC; there was a significant difference in the overall survival rate between the two groups (P = 0.001). (D) No significant differences in 5-year survival rates were found between low levels (n = 45) and high levels (n = 14) of 14-3-3σ expression in ESCC patients with late clinical stage (III–IV, P = 0.328). (E) The 5-year overall survival rates in ESCC patients without lymph node metastasis were 61.28%, 32.54% in high (n = 26) and low levels (n = 71) of 14-3-3σ expression, respectively; there was a significant difference in the overall survival rate between the two groups (P = 0.002). (F) No significant differences in 5-year survival rates were found between low levels (n = 51) and high levels (n = 16) of 14-3-3σ expression in ESCC patients with lymph node metastasis (P = 0.542).
Table 3.
Univariate and multivariate Cox regression analyses of the prognostic variables in ESCC patients.
Figure 4.
Receiver operating characteristic (ROC) curves and Kaplan-Meier survival estimates of evaluated ESCC patients from both (A, C) training and (B, D) validation cohorts.
ROC curves for sex, age, T stage, histological grade, lymph node metastasis, clinical stage, 14-3-3σ and support vector machine (SVM) classifier as predictors for 5-year survival in training (A) and validation (B) cohorts. Kaplan-Meier survival estimates for low- and high-risk ESCC patients as defined by SVM classifier. Overall survival curves of evaluated patients in training (C) and validation (D) cohorts. Log-rank test used to calculate P values.
Table 4.
Clinicopathological features of ESCC patients in the training and validation cohorts.