Proteomic identification of tumor biomarkers associated with primary gallbladder cancer

Figure 1 Representative two-dimensional gel electrophoresis protein profiles. Two-dimensional gel electrophoresis (2-DE) profiles of Primary gallbladder carcinoma (PGC) (A), cholecystitis (B) and normal gallbladder (C) samples. Proteins were separated by pI (X-axis) and molecular mass (Y-axis). Black circles indicate 7 differentially expressed protein spots identified by MALDI-TOF/TOF MS.

Figure 2 Western blot analysis of proteins. Expression of ANXA4, Hsp90β, Dync1h1, ACTG, ACTA2 and α-vinculin in paired gallbladder cancer tissues (Ca) and normal tumor-adjacent tissues (N). Quantification was performed by densitometry of Western blot bands. Densities were normalized to α-vinculin, and relative folds were normalized to expression levels in normal tumor-adjacent tissues. The data are representative of 20 paired groups. ANXA4: Antibody against annexin A4; ACTG: Gamma-actin; ACTA2: Aortic smooth muscle actin; Dync1h1: Dynein cytoplasmic 1 heavy chain 1; Hsp90β: Heat shock protein 90-beta.

Figure 3 mRNA expression levels of biomarkers. The mRNA levels of ANXA4, Hsp90β, Dync1h1, ACTG and ACTA2 in normal tumor-adjacent tissue (N) and cancer tissue (Ca) were measured by quantitative real-time PCR. Data are normalized to expression levels of normal tumor-adjacent tissue. Results are mean ± SE (n = 6). ^bP < 0.01. ANXA4: Antibody against annexin A4; ACTG: Gamma-actin; ACTA2: Aortic smooth muscle actin; Dync1h1: Dynein cytoplasmic 1 heavy chain 1; Hsp90β: Heat shock protein 90-beta.

Figure 4 Immunohistochemical assay of proteins. Representative photographs of the expression levels of ANXA4, Hsp90β and Dync1h1 in paired gallbladder cancer tissues (Ca) and normal tumor-adjacent tissues (N). Scale bar: 100 μm. ANXA4: Antibody against annexin A4; Dync1h1: Dynein cytoplasmic 1 heavy chain 1; Hsp90β: Heat shock protein 90-beta.