Ession in all cases. D) Cytoplasmic T-STAR expression in ER+ cases only. doi:10.1371/journal.pone.0070596.g.75 . The nuclear and cytoplasmic staining intensity was scored as 0 = absent, 1 = weak, 2 = moderate and 3 = strong and 0 = absent, 1 = weak and 2 = moderate, respectively. Examples of tumors with negative and strong staining are shown in Figure 1. No membranous staining was observed and nuclear and cytoplasmicstaining correlated strongly (p,0.001). The dual localizations are in agreement with previous studies where the STAR family member QKI-5 has been found to be 10457188 shuttled between the two compartments [32] and also Sam68 is cytoplasmically expressed in various cancerous tissues [33?5]. In this study, T-STAR expression wasFigure 3. Kaplan-Meier curves correlating T-STAR expression to survival using a 16574785 dichotomized variable where any nuclear staining and intensity have been grouped together. A) Showing T-STAR expression in all cases. B) Showing T-STAR expression in ER+ cases only. doi:10.1371/journal.pone.0070596.gT-STAR Protein Expression in Breast CancerTable 3. Cox uni- and multivariate analysis of Recurrence Free Survival Title Loaded From File according to nuclear T-STAR expression.Covariate categoryUnivariate HR 95 CIMultivariatep-valueHR95 CIp-valueTumor size ,20 mm 20 mm Nodal status neg pos NHG I I III ER Status Positive Negative Age .50 years ,50 years HER2 status 3 0? Adjuvant therapy Treated (chemo/endocrine) Untreated T-STAR nuclear staining negative positive 1.0 0.43 (0.26?.70) 0.001** 1.0 0.48 (0.27?.85) 0.011** 1.0 0.36 (0.25?.53) ,0.001** 1.0 1.3 (0.59?.0) 0.49 1.0 0.98 (0.59?.6) 0.95 1.0 0.98 (0.43?.2) 0.96 1.0 1.4 (0.92?.0) 0.13 1.0 1.1 (0.56?.0) 0.87 1.0 1.7 (1.1?.6) 0.017** 1.0 1.3 (0.64?.0) 0.41 1.0 3.2 (2.3?.4) ,0.001** 1.0 2.1 (1.3?.6) 0.003** 1.0 3.3 (2.4?.7) ,0.001** 1.0 3.4 (1.7?.0) ,0.001** 1.0 2.1 (1.5?.9) ,0.001** 1.0 1.2 (0.73?.0) 0.Note: T-STAR nuclear positive staining is defined as .10 . doi:10.1371/journal.pone.0070596.tsimilar in the in situ- and invasive components in cases where both entities had been sampled. Also in normal glands and ducts, scattered nuclear and sometimes cytoplasmic positivity was seen, seldom exceeding 50 of the cells. Furthermore, correlation between T-STAR and established clinicopathological parameters and tumor markers was investigated and the results are presented in Table 2. Nuclear staining was defined by intensity, but similar associations were seen when the fraction of positive cells was assessed (data not shown). Interestingly, nuclear sub-localization of T-STAR was strongly associated with positive HER2 status and inversely associated with hormone Title Loaded From File receptor positivity. There was also an inverse correlation between T-STAR expression and age at diagnosis. No significant correlations to other clinicopathological parameters were observed. TSTAR expression was also correlated to investigative markers, with both nuclear and cytoplasmic staining being significantly associated with VEGF expression (p = 0.001 and 0.002 respectively) and cytoplasmic, but not nuclear, staining being significantly associated with expression of VEGFR2 (p = 0.004). In line with its inverse relationship to hormone receptor status, nuclear but not cytoplasmic T-STAR expression was inversely associated with cyclin D1 expression (p = 0.001). Of note, there was no significant association with Ki-67. Ki-67 is considered an important prognostic marker for invasive breast cancer [36] but is mainly used in combination with.Ession in all cases. D) Cytoplasmic T-STAR expression in ER+ cases only. doi:10.1371/journal.pone.0070596.g.75 . The nuclear and cytoplasmic staining intensity was scored as 0 = absent, 1 = weak, 2 = moderate and 3 = strong and 0 = absent, 1 = weak and 2 = moderate, respectively. Examples of tumors with negative and strong staining are shown in Figure 1. No membranous staining was observed and nuclear and cytoplasmicstaining correlated strongly (p,0.001). The dual localizations are in agreement with previous studies where the STAR family member QKI-5 has been found to be 10457188 shuttled between the two compartments [32] and also Sam68 is cytoplasmically expressed in various cancerous tissues [33?5]. In this study, T-STAR expression wasFigure 3. Kaplan-Meier curves correlating T-STAR expression to survival using a 16574785 dichotomized variable where any nuclear staining and intensity have been grouped together. A) Showing T-STAR expression in all cases. B) Showing T-STAR expression in ER+ cases only. doi:10.1371/journal.pone.0070596.gT-STAR Protein Expression in Breast CancerTable 3. Cox uni- and multivariate analysis of Recurrence Free Survival according to nuclear T-STAR expression.Covariate categoryUnivariate HR 95 CIMultivariatep-valueHR95 CIp-valueTumor size ,20 mm 20 mm Nodal status neg pos NHG I I III ER Status Positive Negative Age .50 years ,50 years HER2 status 3 0? Adjuvant therapy Treated (chemo/endocrine) Untreated T-STAR nuclear staining negative positive 1.0 0.43 (0.26?.70) 0.001** 1.0 0.48 (0.27?.85) 0.011** 1.0 0.36 (0.25?.53) ,0.001** 1.0 1.3 (0.59?.0) 0.49 1.0 0.98 (0.59?.6) 0.95 1.0 0.98 (0.43?.2) 0.96 1.0 1.4 (0.92?.0) 0.13 1.0 1.1 (0.56?.0) 0.87 1.0 1.7 (1.1?.6) 0.017** 1.0 1.3 (0.64?.0) 0.41 1.0 3.2 (2.3?.4) ,0.001** 1.0 2.1 (1.3?.6) 0.003** 1.0 3.3 (2.4?.7) ,0.001** 1.0 3.4 (1.7?.0) ,0.001** 1.0 2.1 (1.5?.9) ,0.001** 1.0 1.2 (0.73?.0) 0.Note: T-STAR nuclear positive staining is defined as .10 . doi:10.1371/journal.pone.0070596.tsimilar in the in situ- and invasive components in cases where both entities had been sampled. Also in normal glands and ducts, scattered nuclear and sometimes cytoplasmic positivity was seen, seldom exceeding 50 of the cells. Furthermore, correlation between T-STAR and established clinicopathological parameters and tumor markers was investigated and the results are presented in Table 2. Nuclear staining was defined by intensity, but similar associations were seen when the fraction of positive cells was assessed (data not shown). Interestingly, nuclear sub-localization of T-STAR was strongly associated with positive HER2 status and inversely associated with hormone receptor positivity. There was also an inverse correlation between T-STAR expression and age at diagnosis. No significant correlations to other clinicopathological parameters were observed. TSTAR expression was also correlated to investigative markers, with both nuclear and cytoplasmic staining being significantly associated with VEGF expression (p = 0.001 and 0.002 respectively) and cytoplasmic, but not nuclear, staining being significantly associated with expression of VEGFR2 (p = 0.004). In line with its inverse relationship to hormone receptor status, nuclear but not cytoplasmic T-STAR expression was inversely associated with cyclin D1 expression (p = 0.001). Of note, there was no significant association with Ki-67. Ki-67 is considered an important prognostic marker for invasive breast cancer [36] but is mainly used in combination with.
