Figure 1
Expression of ANXA4 in human gallbladder cancer tissues and cell lines.
(A) Representative photographs of immunohistochemical staining of ANXA4 protein (brown) in gallbladder cancer (GBC) tumor and normal tumor-adjacent tissues (N). Original magnification (×100, full; ×400, partial enlargement). (B) Western blot analysis of ANXA4 protein accumulation in gallbladder cancer and normal adjacent tissues. ANXA4 protein levels were significantly elevated in GBC tissues compared with normal adjacent tissues. GAPDH as a positive control. (C) ANXA4 mRNA expression levels in gallbladder cancer tissues (n = 20) were significantly higher than in normal adjacent specimens (n = 20, P < 0.001), according to quantitative RT-PCR. (D) Western blot analysis of ANXA4 and GAPDH protein levels in GBC-SD, SGC-996, NOZ and OCUG-1 gallbladder cancer cell lines. GBC-SD and NOZ cells expressed the highest levels of ANXA4 protein. Human ovarian clear cell adenocarcinoma OVTOKO cells as a positive control. (E) OS of gallbladder cancer patients associated with ANXA4 expression. Kaplan–Meier analysis of the GBC patients, indicated the poorer survival of patients with high ANXA4 expression (χ2 value = 19.371, P < 0.001). The median OS of GBC patients with low ANXA4 expression was 39 (95% confidence interval (CI), 34.7–43.3) months, while the median OS of high-ANXA4-expressing patients was 8 (95% CI, 5.7–10.3) months. The actual 3-year and 5-year OS rates for patients with low ANXA4 expression (63.3% and 33.3%, respectively) were higher than those for patients with high ANXA4 expression (13.3% and 10.0%, respectively, P < 0.001).
