Immunohistochemical staining of WNT10A, β-catenin, and cyclin D1 in kidney tissues.

<p>(A) WNT10A showed very low cytoplasmic expression in renal tubular cells of non-tumoral tissues from both the cortex and medulla. Moreover, WNT10A showed a dramatically higher cytoplasmic expression in RCC tissues. To confirm WNT10A immunohistochemical profiles, WNT10A antibody (sc69135) was used for a serial tissue microarray section, and antibody ablation was performed using its competitive peptide (sc69135P). WNT10A antibody (sc69135) showed similar profiles, and antibody ablation with WNT10A peptide abolished the staining pattern (sc69135P). (B) WNT10A showed higher cytoplasmic expression in RCC cells (A: cortex, B: CCRCC; 400×). β-catenin showed higher intracellular accumulation (cytoplasmic and nuclear; red arrows indicate some representative nuclear-stained cells) in RCC tissues (C: cortex, D: CCRCC; 400×). However, β-catenin showed higher membranous expression in renal tubular cells from non-tumoral normal kidney tissue. Moreover, c-myc (E: cortex, F: CCRCC; 400×) and cyclin D1 (G: cortex, H: CCRCC; 400×) showed higher nuclear expression in RCC tissues, but cyclin D1 showed weak cytoplasmic expression in renal tubular cells from non-tumoral normal kidney tissue. (C) Kaplan-Meier analysis of WNT10A, nuclear β-catenin, and nuclear cyclin D1 accumulative effects on overall survival (OS) and disease-free survival (DFS). There exists an accumulated dose effect, in that patients with higher WNT10A, nuclear β-catenin, and nuclear cyclin D1 levels have poor RCC prognosis (n = initial patient number, s = number of survivors at the end of the study). The 4 groups were defined as follows: 0, carriers with lower expression of all the 3 markers; 1, carriers with higher expression of any 1 of the 3 markers; 2, carriers with higher expression of any 2 of the 3 markers; 3, carriers with higher expression of all the 3 markers. <i>P</i> of log-rank test <0.001.