Expression and clinical role of the bric-a-brac tramtrack broad complex/poxvirus and zinc protein NAC-1 in ovarian carcinoma effusions

We recently identified NAC-1, member of the bric-a-brac tramtrack broad complex/poxvirus and zinc domain family, as an overexpressed gene in ovarian serous carcinoma and found more frequent NAC-1 protein expression in recurrent compared to primary tumors. In the present study, we assessed the clinical significance of NAC-1 expression in ovarian carcinoma effusions. Formalin-fixed, paraffin-embedded sections from 176 effusions (137 peritoneal, 39 pleural) and 197 corresponding solid tumors (69 primary tumors, 128 solid metastases) were analyzed for NAC-1 expression using immunohistochemistry. Staining intensity and extent results were analyzed for possible association with clinicopathologic parameters and survival. Nuclear NAC-1 immunoreactivity was found in carcinoma cells in 98% of (173/176) effusions, 94% (65/69) of primary tumors, and 95% (121/128) of metastases. Staining intensity and extent were significantly higher in effusions compared with matched solid tumors (P = .002 for intensity, P = .003 for extent compared with primary tumors; P < .001 for both intensity and extent compared with metastases). Furthermore, NAC-1 expression intensity was significantly higher in specimens obtained after the administration of chemotherapy (P = .002) and correlated with shorter progression-free survival (PFS) in analysis of 62 patients with post-chemotherapy effusions (P = .039). International Federation of Gynecology and Obstetrics stage (IV versus III) was the only clinical parameter associated with PFS in this group (P = .004). In Cox analysis, only the International Federation of Gynecology and Obstetrics stage was an independent predictor of shorter PFS (P = .009). In conclusion, NAC-1 expression is higher in ovarian carcinoma cells in effusions compared with their solid tumor counterparts. NAC-1 is up-regulated in tumor cells after chemotherapy, suggesting a role for this protein in tumor progression and in the development of chemotherapy resistance in ovarian cancer.     

Expression of Rsf-1, a chromatin-remodeling gene, in ovarian and breast carcinoma

Rsf-1 protein is a member of a chromatin-remodeling complex that plays an important role in regulating gene expression and cell proliferation. Our previous study showed that Rsf-1 was an amplified gene that participated in the development of ovarian serous carcinoma. To further elucidate the role of Rsf-1 in ovarian cancer, we studied Rsf-1 immunoreactivity in 294 ovarian tumors of various histologic types. Because the Rsf-1 amplicon overlaps an amplified region reported in breast cancer, we included 782 neoplastic and normal breast tissues for comparison. Immunohistochemistry was performed on tissue microarrays using a 4-tiered scoring system. Overexpression of Rsf-1 was defined as a nuclear immunointensity of 3+ to 4+ because of a strong correlation between 3+ and 4+ immunointensity and Rsf-1 gene amplification, based on our previous fluorescence in situ hybridization analysis. Rsf-1 overexpression was observed in 25% of high-grade ovarian serous carcinomas and in only rare cases (<7%) of low-grade ovarian serous, ovarian endometrioid, and invasive breast carcinomas but not in any ovarian serous borderline tumors, ovarian clear cell carcinomas, ovarian mucinous carcinomas, intraductal carcinomas of the breast, and normal ovaries and breast tissues. Thus, overexpression of Rsf-1 was significantly associated with high-grade ovarian serous carcinoma (P < .05), as compared with other types of ovarian tumors and breast carcinomas. Our results provide evidence that Rsf-1 expression is primarily confined to high-grade serous carcinoma, the most aggressive ovarian cancer. Because Rsf-1 overexpression occurs in only a small number of breast carcinomas, it is unlikely that Rsf-1 is a critical gene in the development of breast carcinoma.     

Expression of HLA-G in malignant mesothelioma and clinically aggressive breast carcinoma

The aim of the present study was to evaluate HLA-G expression in breast carcinoma and malignant mesothelioma (MM). Malignant breast carcinoma effusions (46) and corresponding solid tumors (39) and 104 MM (26 effusions, 78 solid tumors) were analyzed using immunohistochemistry (IHC). HLA-G protein and mRNA expression were further studied using immunoblotting (IB) and RT-PCR. HLA-ABC expression was analyzed using flow cytometry (FCM). IHC showed predominantly focal HLA-G expression in 12 of 46 (26%) breast carcinoma effusions and 16 of 39 (41%) solid lesions. In MM, 20 of 78 (26%) solid lesions and 14 of 26 (54%) effusions were focally HLA-G positive. Expression in MM was higher in effusions (p=0.008). IB showed more frequent HLA-G expression in MM compared with breast carcinoma effusions, while RT-PCR showed HLA-G mRNA expression in both tumors. FCM showed conserved HLA-ABC expression in 15 of 15 effusions. Breast cancer patients with HLA-G-positive tumor cells had shorter disease-free survival (mean 37 vs 85, median 25 vs 31 months), though not significantly (p=0.14). In conclusion, HLA-G is focally expressed in MM and breast carcinoma, while HLA-ABC expression is conserved. However, the up-regulated expression of HLA-G in MM effusions and its possible association with shorter disease-free survival in advanced stage of breast carcinoma suggest a possible role in immune response evasion in some tumors.     

Rsf-1/HBXAP overexpression is associated with disease-specific survival of patients with gallbladder carcinoma

Chen T‐J, Huang S‐C, Huang H‐Y, Wei Y‐C, Li C‐F. Rsf‐1/HBXAP overexpression is associated with disease‐specific survival of patients with gallbladder carcinoma. APMIS 2011; 119: 808–14. Dysregulated chromatin remodeling often leads to abnormal gene expression or silencing in cells, thereby implicating tumor development and progression. As a subunit of remodeling and spacing factor (RSF) complex, Rsf‐1, a novel nuclear protein with histone chaperon function, mediates ATPase‐dependent chromatin remodeling and confer tumor aggressiveness in common carcinomas. We aimed, for the first time, to evaluate the Rsf‐1 expression status and its associations with clinicopathological features and patient survival in a well characterized cohort of gallbladder carcinomas. Using tissue microarray‐based immunohistochemistry, we assessed Rsf‐1 expression in gallbladder carcinomas, yielding 88 cases undergoing surgical intervention with interpretable results. The Rsf‐1 overexpression, present in 61 cases (69.3%), was significantly associated with higher histological grades (p = 0.002) and vascular invasion (p = 0.037) and marginally with non‐papillary histotypes (p = 0.058). In univariate log‐rank analysis, Rsf‐1 overexpression was significantly predictive of disease‐specific survival (p = 0.0015), which remained prognostically independent [p = 0.0191, risk ratio (RR) = 2.683], along with American Joint Committee on Cancer stages II–IV (p = 0.0265, RR = 2.102). Our findings indicate that Rsf‐1 overexpression is common and associated with adverse prognosticators in gallbladder carcinomas. It may confer tumor aggressiveness through chromatin remodeling and represents a potential prognostic biomarker of gallbladder carcinomas.     

Different clinical roles for p21-activated kinase-1 in primary and recurrent ovarian carcinoma

We recently found amplification of chromosome 11q13.5 in high-grade ovarian serous carcinoma. In the present study, we analyzed the protein expression and clinical significance of p21-activated kinase-1, one of the genes amplified at this site. Formalin-fixed paraffin-embedded sections from 186 effusions (152 peritoneal, 34 pleural) were immunostained. p21-Activated kinase-1 expression in tumor cells was analyzed for possible association with clinicopathologic parameters and survival. The association between protein expression of p21-activated kinase-1 and Rsf-1, a chromatin remodeling protein whose gene colocalizes with p21-activated kinase-1, was additionally studied. p21-Activated kinase-1 protein expression was found in carcinoma cells in 158 (85%) of 186 effusions. Of these, 62 (39%) stained weakly and 96 (61%) strongly. p21-Activated kinase-1 was coexpressed with Rsf-1 (P = .006). Specimens from patients diagnosed with International Federation of Gynecology and Obstetrics stage IV disease had higher staining intensity compared with stage III tumors (P = .014). Univariate survival analysis for patients with primary diagnosis prechemotherapy effusions demonstrated a significant association between higher p21-activated kinase-1 staining extent and longer overall survival (P = .024). In addition, higher staining extent (P = .015) and intensity (P = .013) correlated with better progression-free survival. In contrast, higher p21-activated kinase-1 staining extent correlated with poor overall survival in disease recurrence postchemotherapy effusions (P = .044). In Cox analysis, higher p21-activated kinase-1 staining extent independently correlated with longer progression-free survival (P = .016) and shorter overall survival (.049) in primary diagnosis and disease recurrence effusions, respectively. p21-Activated kinase-1 is frequently expressed in ovarian carcinoma cells in effusions and is associated with opposite prognostic role in primary and recurrent disease. This suggests altered cellular function for this kinase along disease progression, possibly chemotherapy mediated.     

Claudin upregulation in ovarian carcinoma effusions is associated with poor survival

Claudins are tight junction proteins that are highly expressed in ovarian carcinoma (OC). The objective of this study was to analyze the anatomic site-related expression and clinical role of claudins in OC. Effusions (n = 218), corresponding primary tumors (n = 81), and solid metastases (n = 164) (total = 463 tumors) were immunostained for claudin-1, claudin-3, claudin-4, and claudin-7. Results were analyzed for association with anatomic site, clinicopathologic parameters, and survival. All 4 claudins were expressed in >85% of tumors at all anatomic sites. However, staining extent of all except claudin-4 was significantly higher in effusions compared with both primary carcinomas and solid metastases (P < .001). In univariate survival analysis of the entire cohort, higher claudin-3 (P = .038) and claudin-7 (P = .035) expression in effusions correlated with shorter overall survival (OS), with similar results for claudin-7 in analysis of progression-free survival (P = .026). In separate analysis for patients with prechemotherapy effusions, higher claudin-7 expression correlated with shorter OS (P = .045). For patients with postchemotherapy effusions, higher claudin-1 (P = .018) and claudin-3 (P = .009) expression correlated with shorter OS. In multivariate survival analysis of the entire cohort, claudin-7 expression was an independent predictor of poor progression-free survival (P = .017). Claudin-3 independently predicted poor OS for patients with postchemotherapy effusions (P = .012). With the exception of claudin-4, claudins are upregulated in OC effusions compared with solid tumors, in agreement with our previous data for cadherins and integrins in this cancer type, suggesting a prosurvival role for these surface molecules. Claudin-3 and claudin-7 expression in effusions independently predicts poor survival in OC.     

Osteopontin expression in ovarian carcinoma effusions is related to improved clinical outcome

Osteopontin, a soluble protein present in all body fluids, is involved in signaling pathways related to adhesion and extracellular matrix interactions, affecting multiple cellular functions, including inflammation, angiogenesis, and tumor metastasis. We studied osteopontin expression by immunohistochemistry and its clinical relevance in 170 effusions (140 peritoneal, 30 pleural) from women with advanced-stage ovarian carcinoma. Carcinoma cells expressed osteopontin in 126 (74%) of 170 effusions. Osteopontin expression was more frequent in effusions from patients with high-grade tumors (P = .036) but was significantly associated with better debulking at primary surgery (P = .019) and complete response to chemotherapy at diagnosis (P = .021). Osteopontin expression was positively associated to that of the previously studied nuclear factor κB inhibitor IκB (P = .019) and negatively related to expression of the inhibitor of apoptosis family member XIAP (P = .008) and the angiogenic marker endoglin (CD105; P = .018). In univariate survival analysis, the presence of osteopontin in carcinoma cells in primary diagnosis prechemotherapy effusions was associated with longer progression-free survival (P = .037), a finding that did not retain its significance in multivariate Cox analysis. This study demonstrates that osteopontin is frequently expressed in ovarian carcinoma effusions. However, its presence in tumor cells at this anatomical site is unexpectedly associated with less aggressive clinical course, suggesting different and yet undefined biological roles for this protein in serous effusions.     

Survivin, a member of the inhibitors of apoptosis family, is down-regulated in breast carcinoma effusions

We analyzed the expression and prognostic role of inhibitors of apoptosis in breast carcinoma effusions. We used immunoblotting to analyze 22 effusions for XIAP, survivin, and livin expression. Based on immunoblotting results, 49 effusions and 46 corresponding solid tumors were immunostained for XIAP and survivin. Results were analyzed for association with anatomic site, clinicopathologic parameters, and survival. Immunoblotting showed frequent expression of XIAP and survivin and no expression of livin. Carcinoma cells in effusions showed lower survivin immunostaining compared with lymph node metastases (P = .008) and primary carcinomas (P = .041). Higher cytoplasmic survivin expression correlated with poor disease-free survival for patients with postchemotherapy effusions (P = .035). XIAP and survivin, but not livin, are frequently expressed in advanced breast carcinoma. Survivin is down-regulated in effusions compared with solid tumors, possibly in relation to the different cellular economy at this anatomic site. Survivin expression may predict disease-free survival for patients with postchemotherapy effusions.     

Case control study of novel prognostic markers and disease outcome in pregnancy/lactation-associated breast carcinoma

A case control study of pregnancy/lactation associated breast carcinoma (PAC) was conducted on 24 test cases with two controls per case, matching age, tumor grade, tumor size and axillary lymph nodes status. During seven years of this study, 6% of all patients with breast cancer had PAC. In this study, 67% of the test cases showed positive axillary lymph nodes compared to 49% in our series of 315 cases of non-pregnancy/non-lactating women with breast carcinoma (p < 0.05). The expression of nine prognostic markers, i.e. ER, PR, p53, C-erbB-2, EGFR, Cathepsin-D, PCNA, DNA ploidy and S-phase fraction, were studied by immunohistochemistry and flow cytometry. Hormone receptor status showed a statistically significant difference between the two groups, i.e. 29% immunoreactivity in test cases compared to 58% in controls with a p value of 0.007. Among p53, C-erbB-2, EGFR and Cathepsin-D in the test group, only EGFR showed a significant correlation, i.e. 33% immunoreactivity in test cases and 19% immunoreactivity in controls (p < 0.05). Higher PCNA positivity was seen in the test group compared to controls, i.e. 35% in test patients and 28% in controls (p < 0.05). Metastasis to bone and liver was a common feature of test patients as compared to controls (p < 0.05). After a median follow-up of 72 months, there was no significant difference in the overall survival (OS) of test cases and controls as 54% deaths were recorded in test patients and 44% in controls at the end of this study (p > 0.05). In summary, in spite of some significant differences in the expression of few prognostic markers, i.e. ER/PR, EGFR, PCNA and metastatic potential, there was no significant difference in the OS of PAC vs. control group if compared stage for stage.     

NKD1 down-regulation is associated with poor prognosis in breast invasive ductal carcinoma

As a negative modulator of the canonical Wnt signaling pathway, Naked1 (NKD1) is widely expressed in many normal tissues. However, the expression and clinicopathological significance of NKD1 in patients with breast cancer is still unclear. The aim of this study was to evaluate NKD1 expression in breast cancer and to investigate the question of whether reduced expression of NKD1 may have any pathological significance in breast cancer development or progression. In this study, we performed western blotting and immunohistochemistry to evaluate the expression of NKD1 and relevance with clinicopathological factors in the breast invasive ductal carcinoma. Reduction of NKD1 was significantly correlated with lymph node metastasis, histological grade and ER expression in breast cancer. Patients with negative NKD1 expression had significantly lower cumulative postoperative 5 year survival rate than those with positive NKD1 expression. This interpretation is in keeping with the results obtained from our in vitro experiments on MDA-MB-231 cells, we demonstrated that upregulation of NKD1 expression by infect with an adenovirus containing a NKD1 vector significantly reduced the migration of breast cancer cells. These data suggest that NKD1 plays an important role in invasion in human breast cancer and it appears to be a potential prognostic marker for patients with breast cancer.     

Lymphoepithelioma-like carcinoma of the breast

Lymphoepithelioma-like carcinoma (LELC) of the breast is a rare, newly recognized subtype of breast carcinoma. Distinction from medullary carcinoma is important because of the difference in biologic behavior of these two neoplasms and LELC of the breast is regarded as an unusual form of lobular carcinoma. We present the case of a 56-year-old female with a breast mass measuring 2 cm in diameter, which was diagnosed as invasive lobular carcinoma with LELC pattern. This is the ninth case reported in the English literature and to the best of our knowledge the first one with lymph node metastasis.     

CCR7和VEGF-C蛋白与乳腺癌预后之间的关系

目的:探讨趋化因子受体7(CCR7)及血管内皮生长因子C(VEGF-C)蛋白在乳腺癌组织中的表达水平,并分析二者与乳腺癌预后的关系。方法:采用免疫组织化学技术,联合检测CCR7和VEGF-C蛋白分别在乳腺癌组织及正常乳腺组织中的表达差异情况,并分析二者与乳腺癌各相关临床病理特征之间的关系。采用Kaplan-Meier法来评估CCR7及VEGF-C蛋白的异常表达与乳腺癌患者生存期之间的关系。结果:CCR7蛋白在乳腺癌组织(68%)中的阳性表达率高于正常乳腺组织(30%),差异有统计学显著性(P0.01);而VEGF-C蛋白在乳腺癌组织(71%)中的阳性表达率也明显高于正常乳腺组织(24%),差异也有统计学显著性(P0.01)。且在乳腺癌组织中,CCR7与VEGF-C蛋白的表达呈正相关关系(r=0.613,P0.01)。CCR7和VEGF-C蛋白的高表达均与淋巴结转移和TNM分期有关(P0.05),而与年龄、肿瘤大小、雌激素受体和孕激素受体均无关。CCR7及VEGFC蛋白阳性表达者的生存期低于阴性表达者,两组比较差异有统计学显著性(P0.05)。结论:CCR7与VEGF-C的异常高表达可能与乳腺癌预后关系密切,二者可作为判断乳腺癌预后不良的重要指标之一。     

人乳腺癌组织中VEGF-C和iNOS的表达及意义

目的研究人乳腺癌组织中血管内皮生长因子-C(vascular endothelial growth factor-C,VEGF-C)和诱生型一氧化氮合酶(inducible nitiric oxide synthase,iNOS)的表达并探讨两者的相关性。方法采用免疫组织化学SABC法检测50例乳腺癌组织和癌旁正常乳腺组织中VEGF-C和iNOS的表达,并分析其相互关系。结果 VEGF-C和iNOS在乳腺癌组织中均呈强阳性表达,而在癌旁正常组织则不表达。VEGF-C的表达与患者年龄、肿瘤大小、分化程度及临床分期无关(P>0.05),而与淋巴结转移密切相关(P<0.05)。iNOS的表达与患者年龄及肿瘤大小无关(P>0.05),而与分化程度、临床分期及淋巴结转移密切相关(P<0.05)。VEGF-C和iNOS的表达呈正相关(γ=0.43,P<0.05)。结论 VEGF-C和iNOS蛋白在乳腺癌的发生中具有协同作用,两者可能共同参与肿瘤淋巴管的生成。     

Expression of DBC1 and SIRT1 is associated with poor prognosis for breast carcinoma

Recently, it has been reported that SIRT1 and DBC1 may be involved in the development of tumors and predict poor survival in some cancers. However, their exact role is not clear. Therefore, we investigated the expression status and clinical significance of DBC1 and SIRT1 expression in breast carcinomas. We evaluated the immunohistochemical expression of DBC1, SIRT1, and p53 using a 3-mm core from 122 patients with breast cancer for tissue microarray. Positive expression of DBC1 and SIRT1 were seen in 71% and 67% of patients, respectively. In the patients with breast cancer, overall, expression of DBC1 and SIRT1 was significantly associated with distant metastatic relapse and shorter relapse-free survival and overall survival by univariate analysis. Tumor stage and DBC1 and SIRT1 expression were also independent prognostic factors by multivariate analysis. Among the patients who had received chemotherapy, DBC1 and SIRT1 expression was significantly associated with distant metastatic relapse and shorter survival by univariate analysis. DBC1 expression was also associated with distant metastatic relapse and shorter survival in patients who had received endocrine therapy, according to univariate and multivariate analysis. In conclusion, this study shows that expression of DBC1 and SIRT1 is a significant prognostic indicator for breast carcinoma patients.