*3.5. Relationship between Ki-67 Labeling Index and Expression of NME1 and* β*-Catenin*

Ki-67 proliferation index was analyzed to investigate whether the effect of NME1 and β-catenin on cisplatin-resistance might be confounded by different cell proliferation activity. Ki-67 proliferation was not significantly different according to abnormal expression of NME1 or β-catenin irrespective of histology (Figure 4A) and cisplatin-based adjuvant chemotherapy (Figure 4B). These observations suggest that NME1 and β-catenin may affect cisplatin-based adjuvant chemotherapy through other mechanisms rather than through any changes in cell proliferation.

**Figure 4.** Ki-67 (MKI67) proliferation index according to altered expression of NME1 and β-catenin. Ki-67 proliferation was compared according to expression statuses of NME1 and β-catenin, stratified by histology (**A**) and adjuvant chemotherapy (**B**). "Adenoca" and "Squamous" represent adenocarcinoma and squamous cell carcinoma, respectively. "No" and "Yes" indicate the absence and presence of altered expression, respectively. *p*-values were calculated using one-way ANOVA.

### *3.6. The Relationship between NME1 and Nuclear* β*-Catenin Expression*

*NME1* silencing is known to induce a redistribution of β-catenin from the cell surface into the cytoplasm and nucleus by activating the Wnt pathway [17]. To understand whether NME1 expression affects the nuclear translocation of β-catenin, we analyzed the levels of nuclear β-catenin expression (Figure 5A) according to NME1 expression. The levels of nuclear β-catenin expression showed a linear relationship with those of cytoplasmic β-catenin expression (Pearson's correlation coefficient γ = 0.74, *p* = 0.002; Figure 5B) but not to those of NME1 expression (γ = −0.03, *p* = 0.80; Figure 5C).

**Figure 5.** The relationship between NME1 and nuclear β-catenin expression. (**A**) β-catenin expression (scale bar, 100 μm) are shown in nucleus. (**B**,**C**) X- and *Y*-axis scores were obtained by multiplying the intensity score of staining with the proportion of positive stained cells. The linear relationship between two variables was calculated using Pearson's correlation coefficient.
