Expression of the p53 Target Wig-1 Is Associated with HPV Status and Patient Survival in Cervical Carcinoma

<div><p>The p53 target gene <i>WIG-1</i> (<i>ZMAT3</i>) is located in chromosomal region 3q26, that is frequently amplified in human tumors, including cervical cancer. We have examined the status of <i>WIG-1</i> and the encoded Wig-1 protein in cervical carcinoma cell lines and tumor tissue samples. Our analysis of eight cervical cancer lines (Ca Ski, ME-180, MS751, SiHa, SW756, C-4I, C-33A, and HT-3) by spectral karyotype, comparative genomic hybridization and Southern blotting revealed <i>WIG-1</i> is not the primary target for chromosome 3 gains. However, <i>WIG-1</i>/Wig-1 were readily expressed and <i>WIG-1</i> mRNA expression was higher in the two HPV-negative cervical cell lines (C33-A, HT-3) than in HPV-positive lines. We then assessed Wig-1 expression by immunohistochemistry in 38 cervical tumor samples. We found higher nuclear Wig-1 expression levels in HPV-negative compared to HPV positive cases (<i>p</i> = 0.002) and in adenocarcinomas as compared to squamous cell lesions (<i>p</i><0.0001). Cases with moderate nuclear Wig-1 staining and positive cytoplasmic Wig-1 staining showed longer survival than patients with strong nuclear and negative cytoplasmic staining (<i>p</i> = 0.042). Nuclear Wig-1 expression levels were positively associated with age at diagnosis (<i>p</i> = 0.023) and histologic grade (<i>p</i> = 0.034). These results are consistent with a growth-promoting and/or anti-cell death function of nuclear Wig-1 and suggest that Wig-1 expression can serve as a prognostic marker in cervical carcinoma.</p></div

Wig-1 expression in cervical carcinomas.

<p>Immunohistochemical (IHC) analysis revealed four main Wig-1 staining patterns: (A) moderate nuclear and positive cytoplasmic Wig-1 staining (IHC staining picture of case CIS-3 as a representative) at low magnification (left) and high magnification (right); (B) strong nuclear and negative cytoplasmic Wig-1 staining (IHC staining picture of case ADCA-18 as a representative) at low magnification (left) and high magnification (right); (C) moderate nuclear and negative cytoplasmic Wig-1 staining (IHC staining picture of case ADCA-21 as a representative) at low magnification (left) and high magnification (right); and (D) strong nuclear and positive cytoplasmic Wig-1 staining (IHC staining picture of case ADCA-22 as a representative) at low magnification (left) and high magnification (right). Neoplasia and non-neoplasia are marked in all figures. Scale bar  = 50 µm.</p

HPV status and chromosome 3 aberrations in the cell lines studied.

<p>* HPV-39 according to ATCC;</p><p>** HPV type was undetermined after DNA sequencing.</p><p>*** Bold in parenthesis indicates amplification within the gained interval.</p><p>HPV status and chromosome 3 aberrations in the cell lines studied.</p

Comparison of Wig-1 IHC expression with clinical parameters.

<p>CIN3  =  Cervical intraepithelial neoplasia 3; CIS  =  Cancer <i>in situ</i>.</p><p>Univariate analysis of variance were performed. P-values <0.05 were regarded as sgnificant and are indicated in bold.</p><p>Comparison of Wig-1 IHC expression with clinical parameters.</p

Copy number and expression of <i>WIG-1/</i>Wig-1 in relation to HPV infection and <i>TP53/p53</i> status.

<p>-  =  negative;</p><p>+  =  detectable to moderate expression;</p><p>++  =  strong expression;</p><p>n.d.  =  not determined;</p><p>n.a.  =  not available;</p><p>* Given in arbitrary units as compared to fibroblasts (1.0).</p><p>** Reiss <i>et al.</i>, 1992; Yaginuma <i>et al.</i>, 1991; Srivastava <i>et al.</i>, 1992; Scheffner <i>et al.</i>, 1991; Masuda <i>et al.</i>, 1987.</p><p>Copy number and expression of <i>WIG-1/</i>Wig-1 in relation to HPV infection and <i>TP53/p53</i> status.</p

Kaplan-Meier survival curves showing cervical carcinoma patient survival as a function of Wig-1 staining pattern.

<p>(A) Patients with moderate nuclear Wig-1 staining show better survival than patients with high nuclear staining. (B) Patients with positive cytoplasmic Wig-1 staining have better survival than patients with negative cytoplasmic staining. (C) Patients with moderate nuclear and positive cytoplasmic Wig-1 staining show better survival than patients with strong nuclear and negative cytoplasmic staining. Cases no. and Log Rank P values are indicated. <i>p</i><0.05 is considered to be significant.</p

Quantitative Proteomics Profiling of Primary Lung Adenocarcinoma Tumors Reveals Functional Perturbations in Tumor Metabolism

In this study, we have analyzed human primary lung adenocarcinoma tumors using global mass spectrometry to elucidate the biological mechanisms behind relapse post surgery. In total, we identified over 3000 proteins with high confidence. Supervised multivariate analysis was used to select 132 proteins separating the prognostic groups. Based on in-depth bioinformatics analysis, we hypothesized that the tumors with poor prognosis had a higher glycolytic activity and HIF activation. By measuring the bioenergetic cellular index of the tumors, we could detect a higher dependency of glycolysis among the tumors with poor prognosis. Further, we could also detect an up-regulation of HIF1α mRNA expression in tumors with early relapse. Finally, we selected three proteins that were upregulated in the poor prognosis group (cathepsin D, ENO1, and VDAC1) to confirm that the proteins indeed originated from the tumor and not from a stromal or inflammatory component. Overall, these findings show how in-depth analysis of clinical material can lead to an increased understanding of the molecular mechanisms behind tumor progression