Cox regression analysis to determine prognostic significance of <i>p53</i> mutations.

1<p>Adjusted for the <i>p53</i> mutations, age, gender, tumor location, tumor grade, tumor size, tumor depth of invasion, nodal involvement, and adjuvant therapy.</p

IHC expression patterns of up-regulated genes in <i>p53</i> mutant phenotypes of MS CRCs.

<p><b>a</b>, normal colonic epithelium (NCE) demonstrating moderate cytoplasmic (thick arrow) and weak membrane (thin arrow) FUT3 staining (400 µm). <b>b</b>, CRCs exhibiting strong cytoplasmic (thick arrows) and moderate to weak membrane FUT3 staining (thin arrows) (400 µm). <b>c</b>, CRCs with weak cytoplasmic staining (thin arrows) (400 µm). <b>d</b>, NCE demonstrating weak cytoplasmic TRIM29 staining with a focal punctuate pattern (thin arrows) (400 µm). <b>e</b>, CRCs exhibiting moderate to strong cytoplasmic TRIM29 staining with a punctate pattern on the luminal aspect (thin arrows) (600 µm). <b>f</b>, CRCs with weak cytoplasmic TRIM29 staining (thin arrows) (400 µm). <b>g</b>, NCE demonstrating weak cytoplasmic to complete lack of IQGAP3 staining. [Note: Lymphocytes in the stroma show moderate cytoplasmic staining (thin arrows); the adjacent tumor demonstrates moderate cytoplasmic and nuclear immunostaining (thick arrows) (400 µm)]. <b>h</b>, CRCs exhibiting strong cytoplasmic IQGAP3 staining (thick arrows) (400 µm). <b>i</b>, CRCs with lack of staining for IQGAP3 (thick arrows) (400 µm). <b>j</b>, NCE demonstrating moderate cytoplasmic staining of SLC6A8 staining (thin arrows) (600 µm). <b>k</b>, CRCs exhibiting strong cytoplasmic SLC6A8 staining with luminal accentuation (thick arrows) (600 µm). <b>l</b>, CRCs negative for SLC6A8 staining (thick arrows) (600 µm).</p

The prognostic significance of <i>p53</i> mutations in survival of Stage III CRC patients with MSS or MSI-H phenotypes (Kaplan-Meier survival curves).

<p><i>p53</i> mutations were associated with worse patient survival (log-rank, <i>P</i> = 0.025) (<b>A</b>) than wt-<i>p53</i> in the subset of MSS phenotype, but not in the subset of MSI-H phenotype (log-rank, <i>P</i> = 0.695) (<b>B</b>).</p

IHC expression patterns of down-regulated genes in <i>p53</i> mutant phenotypes of MS CRCs.

<p><b>a</b>, NCE demonstrating moderate nuclear and weak cytoplasmic PDLIM3 staining [Note: Lymphocytes in the stroma and the normal epithelial cells show moderate nuclear staining (thin arrows); the adjacent tumor demonstrates strong nuclear and moderate cytoplasmic immunostaining (thick arrows) (400 µm)]. <b>b</b>, CRCs exhibiting lack of PDLIM3 immunostaining (thin arrows) (600 µm). <b>c</b>, CRCs with strong nuclear and weak cytoplasmic PDLIM3 staining (thick arrows) (600 µm). <b>d</b>, NCE demonstrating weak cytoplasmic LPAR6 staining (thin arrows) (600 µm). <b>e</b>, CRCs exhibiting focal weak cytoplasmic LPAR6 staining (thin arrows) (600 µm). <b>f</b>, CRCs with weak to moderate cytoplasmic (thick arrows) and focal nuclear LPAR6 staining (thin arrows) (600 µm). <b>g</b>, NCE demonstrating moderate to strong cytoplasmic PLAT staining (thin arrows) (600 µm). <b>h</b>, CRCs exhibiting focal moderate cytoplasmic PLAT staining (thick arrows) (600 µm). <b>i</b>, CRCs with moderate to strong staining for PLAT (thick arrows) (600 µm).</p

Gene expression profiles of CRCs based on <i>p53</i> status.

<p>Genes and samples were clustered independently by hierarchical clustering. Rows represent genes, and columns represent samples, which are color-coded by <i>p53</i> status (blue and red correspond to wt-<i>p53</i> and <i>p53</i> mutant, respectively. The color scale is shown at bottom right. Values are expressed as log2-ratios of expression in CRCs with <i>p53</i> mutant phenotypes to that in wt-<i>p53</i> phenotypes. (A & B) Clusters of 35 and 49 genes showing consistent up-regulation and down-regulation in <i>p53</i> mutant phenotypes.</p

Validation of deferentially expressed genes by qNPA and qRT-PCR based on <i>p53</i> status in Stage III-MSS CRCs.

<p>Abbreviations: Ch, chromosome number; AHG, Affymetrix human genome plus 2.0 array.</p>1<p>Fold change in differential expression of genes by AHG;</p>2<p>Fold change by qNPA;</p>3<p>Fold change qRT-PCR.</p