Immunostaining results for p15^INK4b and p57^KIP2.

<p>Immunostaining results for p15^INK4b and p57^KIP2.</p

Survival curves using the Kaplan-Meier method for a subgroup of patients obtained surgical radicality (no rest tumor).

<p>The Kaplan-Meier curve of disease-specific survival in relation to the combined analysis of p14^ARF/p15^INK4b/p16^INK4a showed that patients whose tumors expressed low levels of two or three of these INK4 proteins had a worse prognosis than those with only low levels of one or no protein (<i>p</i> = 0.06).</p

p15^INK4b and p57^KIP2 expression in relation to clinicopathological variables.

1<p>Linear-by-linear association.</p>2<p>Pearson chi-square.</p><p>High: Expression in nucleus = 9.</p><p>Low: Expression in nucleus <9.</p

Survival curves using the Kaplan-Meier method.

<p>The Kaplan-Meier curve of disease-specific survival in relation to the combined analysis of p14^ARF/p15^INK4b/p16^INK4a showed that patients whose tumors expressed low levels of two or three of these INK4 proteins had a worse prognosis than those with only low levels of one or no protein (<i>p</i> = 0.02).</p

Immunohistochemical staining of p15^INK4b and p57^KIP2 protein in vulvar squamous epithelium.

<p>High nuclear expression of p15^INK4b (A) and p57^KIP2 (B) in normal vulvar epithelium. High nuclear expression of p15^INK4b (C) and p57^KIP2 (D) and low nuclear expression of p15^INK4b (E) and p57^KIP2 (F) in vulvar carcinomas.</p

IHC SHBG staining in clinical ovarian carcinoma samples.

<p>Representative weak (A), moderate (B, moderately differentiated tumor) and strong (C, poorly differentiated tumor) are shown. Stronger SHBG immunostaining is shown at the invasive front of a poor-differentiated ovarian carcinoma (D). Negative control performed in human liver tissue with the same concentration of non-immune goat IgG shows no staining in the liver cells (E). Positive control performed with liver tissue shows the positive cytoplasm SHBG staining. Endothelial cells are also positive for SHBG. All photos were taken at 200×.</p

Survival curves using the Kaplan-Meier method (288 VSCC without pre-surgery treatment cases).

<p>The Kaplan-Meier curves of disease-specific survival in relation to combinations of (A) (pCDK1^{Thr161 C+N} + 14-3-3σ^N), (B) (pCDK1^{Thr161 C+N} + 14-3-3η^C), (C) (pCDK1^{Thr161 C+N} + Wee1^C) and (D) (pCDK1^{Thr161 C+N} + 14-3-3σ^N + 14-3-3η^C + Wee1^C). The <i>p</i>-values differ slightly from those in <a href="http://www.plosone.org/article/info:doi/10.1371/journal.pone.0121398#pone.0121398.t002" target="_blank">Table 2</a> due to the use of the log-rank test as opposed to the Cox-regression analysis.</p

Comparison of clinical and pathologic characteristics by tumor SHBG intensity.

<p>Muci, Mucinous tumor; Endo, Endometrioid; Clea, Clear cell; Mixe, Mixed epithelial tumor; Undi, Undifferenciated tumor.</p

OS and PFS for the ovarian carcinoma patients in 3 SHBG IHC groups.

<p>For both OS (A) and PFS (B) curves, weakly (IHC score 1), moderately (IHC score 2) and strongly (IHC score 3) SHBG expressing groups were marked by blue, green and brown lines respectively. No statistically significant difference was obtained both for OS (<i>p</i> = 0.220) and PFS (<i>p</i> = 0.132).</p

Expression of CDK1^Tyr15, pCDK1^Thr161, Cyclin B1 (Total) and pCyclin B1^Ser126 in Vulvar Squamous Cell Carcinoma and Their Relations with Clinicopatological Features and Prognosis

<div><p>Cyclin B1-CDK1 complex plays an important role in the regulation of cell cycle. Activation of Cyclin B1 and CDK1 and the formation of the complex in G2/M are under multiple regulations involving many regulators such as isoforms of 14-3-3 and CDC25 and Wee1. Abnormal expression of Cyclin B1 and CDK1 has been detected in various tumors. However, to our knowledge no previous study has investigated Cyclin B1 and CDK1 in vulvar cancer. Therefore, we evaluated the statuses of CDK1^Tyr15, pCDK1^Thr161, Cyclin B1 (total) and pCyclin B1^Ser126 in 297 cases of vulvar squamous cell carcinomas by immunohistochemistry. Statistical analyses were performed to explore their clinicopathological and prognostic values. In at least 25% of tumor cases high expression of CDK1^Tyr15, pCDK1^Thr161, Cyclin B1 (total) and pCyclin B1^Ser126 was observed, compared to the low levels in normal vulvar squamous epithelium. Elevated levels of CDK1^Tyr15, pCDK1^Thr161, Cyclin B1 (total) and pCyclin B1^Ser126 were correlated with advanced tumor behaviors and aggressive features. Although CDK1^Tyr15, pCDK1^Thr161, Cyclin B1 (total) and pCyclin B1^Ser126 could not be identified as prognostic factors, combinations of (pCDK1^{Thr161 C+N} + 14-3-3σ^N), (pCDK1^{Thr161 C+N} + 14-3-3η^C), (pCDK1^{Thr161 C+N} + Wee1^C) and (pCDK1^{Thr161 C+N} + 14-3-3σ^N + 14-3-3η^C + Wee1^C) were correlated with disease-specific survival (<i>p</i> = 0.036, <i>p</i> = 0.029, <i>p</i> = 0.042 and <i>p</i> = 0.007, respectively) in univariate analysis. The independent prognostic significance of (pCDK1^{Thr161 C+N} + 14-3-3σ^N + 14-3-3η^C + Wee1^C) was confirmed by multivariate analysis. In conclusion, CDK1^Tyr15, pCDK1^Thr161, Cyclin B1 (total) and pCyclin B1^Ser126 may be involved in progression of vulvar squamous cell carcinoma. The combination of pCDK1^Thr161, 14-3-3σ, 14-3-3η and Wee1 was a statistically independent prognostic factor.</p></div