Copy number changes of target genes in chromosome 3q25.3-qter of esophageal squamous cell carcinoma: TP63 is amplified in early carcinogenesis but down-regulated as disease progressed

Figure 1 Immunoreactivity for TP63. A: Intense nuclear TP63 staining; B: Weak nuclear TP63 staining; C: Negative staining. (Original magnification: ×400).

Figure 2 Comparison of copy numbers assessed by FISH (stripped bar) and Q-PCR (±SD) (hollow bar) of 13 genes in 4 EC-SCC cell lines [CE 48T/VGH (pink), CE 81T/VGH (yellow), TE6 (green), TE9 (blue)]. Pearson coefficients of correlation for comparing FISH and Q-PCR of 13 genes in 4 cell lines were all >0.9.

Figure 3 Average copy number changes (indicated by the circles with number listed beside them; crossbar represented 2×standard error) of 13 genes in association of their relative position on chromosome 3q. Four clusters of amplifications could be found: SSR3 and CCLN1 at 3q25.31; hTERC and SKIL at 3q26.2; ECT2 and PIK3CA at 3q26.31-q26.32; and SST, TP63 and TFRC at 3q27.3-q29.

Figure 4 In Q-PCR analysis of 60 EC-SCC tumors, patients with lymph node metastasis [LN (+)] have significantly lower copy number of TP63 in the primary tumor than those without lymph node metastasis [LN (-)] (2. 05 vs 3.08; P = 0.017).

Figure 5 In IHC study of 111 EC-SCC tumors, patients with lymph node metastasis [LN (+)] have significantly lower TP63 staining score in the primary tumor than those without lymph node metastasis [LN (-)] (51. 03 vs 79.62; P = 0.034).