Expression of BRAF V600E mutant protein in epithelial ovarian tumors.

BACKGROUND: Genetic analyses have identified BRAF V600E mutations in a subset of ovarian carcinomas. The aim of this study was to investigate the expression of BRAF V600E aberrant protein using a novel mutation-specific antibody in epithelial ovarian tumors. METHODS: We immunohistochemically analyzed expression of V600E-mutant BRAF protein in archival formalin-fixed, paraffin-embedded tissue specimens of 142 epithelial ovarian tumors [98 invasive carcinomas and 44 tumors of low malignant potential (LMP)] using monoclonal antibody VE1. BRAF mutation status was validated in all immunopositive cases and in 6 immunonegative control cases by gene sequencing. RESULTS: We found anti-BRAF V600E immunolabeling in 4 serous carcinomas and 5 serous LMP. Presence of a BRAF V600E mutation was confirmed by sequencing analysis in 6 of the 9 cases (3 LMP tumors, 3 low-grade carcinomas). In 2 partially VE1-positive tumors deriving from 1 patient (1 LMP and 1 contralateral invasive high-grade serous carcinoma), genetic analysis repeatedly resulted in BRAF wild-type, arguing for false-positive immunostaining results. One immunopositive case was repeatedly inconclusive in genetic analysis. In all 6 genetically confirmed cases, BRAF V600E mutant protein expression was homogenous throughout the tumor tissue. CONCLUSIONS: We found BRAF V600E mutations in 13% (4/31) of serous LMP and 5% (3/62) of invasive serous carcinomas. No BRAF V600E mutations were detected in nonserous epithelial ovarian tumors. For reliable assessment of the BRAF V600E status in ovarian epithelial tumor samples, an integrated approach using immunohistochemistry and genetic analysis seems advisable, as both methods lead to incorrect results in some cases.

Human homologue for Caenorhabditis elegans CUL-4 protein overexpression is associated with malignant potential of epithelial ovarian tumours and poor outcome in carcinoma.

BACKGROUND: CUL-4 plays a critical role in DNA replication in Caenorhabditis elegans, and interacts with p53 and p21 proteins in cell cycle regulation and response to genomic instability. However, the role of CUL-4 in human carcinomas is widely unknown. AIMS: To investigate the expression of CUL-4 protein and its association with p53 and p21, and to determine its prognostic relevance in invasive ovarian carcinoma. METHODS: CUL-4, p53 and p21 protein expression was determined retrospectively by immunohistochemistry in 140 specimens of human epithelial ovarian tumours (98 invasive carcinomas and 42 tumours of low malignant potential; LMP). RESULTS: Overexpression of CUL-4 was observed in 41 (41.8%) of carcinoma samples and in 10 (23.8%) LMP tumours. CUL-4 was significantly more often overexpressed in invasive carcinomas compared with LMP tumours (p=0.042, χ(2) test, OR 2.302, 95% CI 1.018 to 5.203). In invasive carcinoma, CUL-4 overexpression was found to be a prognostic factor for overall (p=0.017, Cox regression, HR 2.387, 95% CI 1.17 to 4.869) and disease-free survival (p=0.005, Cox regression, HR 3.5, 95% CI 1.465 to 8.365), respectively. In subgroup analysis, CUL-4 was only of prognostic relevance in carcinomas without p53 expression. CONCLUSION: These data indicate for the first time that CUL-4 might play a relevant role in the development and progression of ovarian carcinoma, warranting further investigations. Degradation of wild-type p53 might be a key mechanism to explain why CUL-4 leads to more aggressive clinical behaviour. Not only CUL-4 itself, but also its associated proteins might represent targets for novel, selective therapeutic strategies.

Overexpression of the human homologue for Caenorhabditis elegans cul-4 gene is associated with poor outcome in node-negative breast cancer.

BACKGROUND: Cul-4, a member of the Caenorhabditis elegans "cullin" ubiquitin-ligase gene family, plays a critical role in regulation of DNA-replication in this nematode. It has been suggested that cul-4 might have an important role in the development and progression of human cancer, but no data on this subject exist. The aim of this study was to investigate the expression and prognostic relevance of CUL-4 protein in lymph node-negative breast cancer, one of the most common malignancies worldwide. MATERIALS AND METHODS: CUL-4 protein expression was determined with immunohistochemistry in 167 specimens of human node-negative invasive breast cancer with long-term follow-up. Results were correlated with overall and disease-free survival of patients. RESULTS: Strong expression of CUL-4 protein was observed in 32 cases (19.2%), moderate expression in 59 (35.3%), weak expression in 64 (38.3%), and in 12 tumors (7.2%) no expression of CUL4 was observed. Patients with strong expression of CUL4 had a significantly shorter overall and disease-free survival (p = 0.04 and p = 0.029, respectively; Cox regression) compared to all other cases. CONCLUSION: Our data provide evidence for the first time that CUL-4 could play an important role in the development and progression of human cancer.

Hypoxia inducible factor-1alpha correlates with VEGF-C expression and lymphangiogenesis in breast cancer.

The transcription factor Hypoxia inducible factor-1alpha (HIF-1alpha) plays a crucial role in tumor progression by regulating angiogenesis, cell survival and drug resistance. HIF-1alpha is also implicated in biological functions under normoxic conditions and recent data provide evidence for a possible role in tumor lymphangiogenesis by regulating the lymphatic vascular endothelial growth factor-C (VEGF-C). In breast cancer, lymphatic vessel invasion (LVI) by tumor cells and subsequent metastasis to axillary lymph nodes is a critical point in progression of the disease with severe therapeutical and prognostic implications. Aim of this study is to investigate the role of HIF-1alpha in VEGF-C expression, lymphangiogenesis, and LVI in lymph node positive breast cancer. Lymphatic microvessel density (LMVD), LVI, HIF-1alpha and VEGF-C protein-expression were evaluated by immunohistochemistry in 119 cases of lymph node positive invasive breast cancer. There was a significant correlation between HIF-1alpha and VEGF-C (p = 0.026, r = 0.204, Spearman's coefficient of correlation). Further a significant association between HIF-1alpha-expression and the amount of peritumoral lymphangiogenesis LMVD was seen (p = 0.014, Mann-Whitney test). LMVD correlated significantly with LVI (p<0.001, Mann-Whitney test). HIF-1alpha was an independent prognostic factor for overall and disease free survival in uni- and multivariate analysis (p = 0.027, 0.029, 0.025, respectively, Cox regression). Our data provide evidence for a possible role of HIF-1alpha as regulator of tumor-associated lymphangiogenesis in human breast cancer and emphasizes the promising status of HIF-1alpha as a therapeutical target against tumor progression and metastasis.

Elective cesarean section vs. spontaneous delivery: a comparative study of birth experience.

BACKGROUND: To investigate birth experience and medical outcome in women with elective cesarean section (CS) compared with women with intended vaginal delivery. METHODS: A total of 1050 pregnant women were included in this prospective trial. Psychological factors, pain levels and birth experience were investigated using a self-designed questionnaire and three established psychological tests in gestational week 38, and 3 days and 4 months postpartum. In addition, medical data were evaluated from the records. RESULTS: Out of 903 women with planned vaginal birth, in 484 women (53.6%) minimal perineal surgery had to be performed after birth, 41 women (4.5%) had vacuum deliveries, and in 93 cases (10.3%) emergency CS had to be performed. In the 147 elective CS (103 based on medical and 44 on psychological factors), a significantly lower rate of maternal and fetal complications was observed when compared with vaginal birth (5.4% vs. 19.3%; p < 0.0001). Birth experience (Salmon test) was significantly better in elective CS compared with vaginal delivery, but worse in women with emergency CS and worst in those with vacuum delivery. We found that 83.5% of women with vaginal delivery would choose the same mode of birth again, 74.3% of women with CS on demand, and 66% of women with medically necessary CS. Only 30.1% of women with emergency CS wanted to receive CS at the next birth. CONCLUSIONS: Elective CS is a safe and psychologically well tolerated procedure. The results are comparable with uncomplicated vaginal delivery and far superior to secondary intervention such as vacuum delivery or emergency CS.

Overexpression of hypoxia-inducible factor 1alpha indicates diminished response to radiotherapy and unfavorable prognosis in patients receiving radical radiotherapy for cervical cancer.

PURPOSE: The purpose is to investigate the impact of hypoxia-inducible factor (HIF)-1alpha expression on response to radiotherapy and prognosis of patients with primary irradiated cervical cancer. Because human papillomavirus (HPV) oncoprotein E6 might interact with HIF-1alpha in various pathways, we also investigated the relation of HIF-1alpha and HPV status. EXPERIMENTAL DESIGN: Expression of HIF-1alpha was investigated by immunohistochemistry in 67 specimens of patients who had received radical radiotherapy for cervical cancer stages IB-IIIB. HPV analysis was performed using type-specific PCR, cloning, and sequencing. Survival analysis was performed using univariate and multivariate analysis. RESULTS: Immunohistochemistry revealed expression of HIF-1alpha in 72.1% of the tumor samples. In 16 (23.9%) cases, there was a weak expression, in 25 (37.3%) a moderate expression, and in 7 cases (10.4%) a strong expression of HIF-1alpha. Nineteen samples (28.4%) were considered negative for HIF-1alpha expression. Strong/moderate expression of HIF-1alpha was associated with only partial response to radiotherapy (P = 0.037, chi(2) test). Strong/moderate expression of HIF-1alpha was also an independent prognostic factor for shorter progression-free survival (P = 0.036, Cox regression) and cervical cancer-specific survival (P = 0.04, Cox regression). No association between HIF-1alpha expression and infection with different HPV types could be found. CONCLUSIONS: Overexpression of HIF-1alpha has predictive and prognostic significance in cervical cancer patients receiving curative radiation therapy. Possibly, expression of HIF-1alpha could serve as intrinsic marker of hypoxia in cervical cancer.

Overexpression of Id-1 is associated with poor clinical outcome in node negative breast cancer.

Id-1 is an important regulator of cellular growth and differentiation and controls malignant progression of breast cancer cells. The aim of our study was to assess the clinical impact of Id-1 expression in breast cancer, i.e., its potential impact on prognosis and prediction of treatment response. Id-1 protein expression was determined immunohistochemically in 191 patients with lymph-node negative breast cancer, and univariate and multivariate survival analysis was carried out. Fifteen (7.9%) specimens showed strong expression, 75 (39.3%) moderate, 55 (28.8%) weak expression and 46 (24.1%) cases no expression of Id-1. Patients with strong or moderate Id-1 expression had a significant shorter overall (p = 0.003, Cox regression) and disease-free survival (p = 0.01, Cox regression) compared to those with absent or low expression. Progesterone receptor density was significantly higher in breast cancers with absent/low Id-1 expression compared to those with moderate/strong expression (p < 0.001, t-test). Id-1 expression was significantly stronger in cases positive for p16(INK4a) expression compared to those negative for p16 (p = 0.049, Mann-Whitney test). The influence of Id-1 on clinical outcome seems much stronger in patients with negative estrogen receptor status compared to those with positive status, who received receptor antagonists as adjuvant therapy in most cases. Overexpression of Id-1 protein represents a strong independent prognostic marker in node negative breast cancer, and future therapies inhibiting Id-1 expression might be beneficial for these patients. Our results also suggest that due to the apparent interaction of Id-1 with the steroid-receptor system in breast cancer, hormonal therapies might influence Id-1 expression and its impact on clinical outcome.

Level of Id-1 protein expression correlates with poor differentiation, enhanced malignant potential, and more aggressive clinical behavior of epithelial ovarian tumors.

PURPOSE: Id (inhibitor of differentiation/DNA binding) -1 is involved in neoangiogenesis, it antagonizes basic helix-loop-helix proteins, inhibits differentiation, and enhances cell proliferation. Aim of this study was to investigate Id-1 protein expression in epithelial ovarian tumors and its clinical relevance in ovarian cancer. EXPERIMENTAL DESIGN: We have investigated Id-1 expression by reverse transcription-PCR and Western blotting in ovarian cancer samples. On the basis of these results, Id-1 protein expression was determined by immunohistochemistry in 101 specimens of epithelial ovarian cancer, in 40 borderline tumors, and in 20 cystadenomas. In these cases, Id-1 expression was correlated with p21 expression, microvessel density, and survival. RESULTS: By immunohistochemistry, detectable expression of Id-1 was found significantly more often in ovarian cancers (74.3%) than in borderline tumors (32.5%) and cystadenomas (0%; P < 0.0001, chi(2) test). Cancer samples with poor or moderate histological differentiation (G3/G2) showed significantly stronger Id-1 expression than cancer samples with high differentiation (G1; P = 0.021, Mann-Whitney test), and no association of Id-1 with p21 expression or microvessel density was found. In cancer samples strong or moderate expression of Id-1 was a strong predictor for shorter overall survival in uni- and multivariate analysis (P = 0.001, Cox-regression). CONCLUSIONS: The level of Id-1 protein expression correlates with the malignant potential of ovarian tumors. In cancer samples, stronger Id-1 expression is associated with poor differentiation and more aggressive behavior of tumor cells, resulting in poor clinical outcome. Consequently, Id-1 inhibition in the future might be of benefit for patients with ovarian cancer.

Overexpression of hypoxia-inducible factor 1alpha is associated with an unfavorable prognosis in lymph node-positive breast cancer.

PURPOSE: Hypoxia-inducible factor (HIF)-1alpha is a transcription factor that supports the adaptation of human cancer cells to hypoxia and is involved in various pathways supporting tumor growth and progression. The aim of this study was to determine the prognostic influence of HIF-1alpha expression in patients with advanced-stage breast cancer, evident by positive lymph nodes. EXPERIMENTAL DESIGN: Expression of HIF-1alpha was determined immunohistochemically in 206 patients with lymph node-positive breast cancer. Furthermore, the interrelationship of HIF-1alpha with p53 and HER-2 protein expression, estrogen receptor density, and survival was analyzed. Colocalization of p53 and HIF-1alpha proteins was analyzed by confocal laser scanning microscopy. RESULTS: Strong nuclear expression of HIF-1alpha by invasive cancer cells was found in 48 patients (23.3%), moderate expression was found in 74 patients (35.9%), and weak expression was found in 35 patients (17%); no expression was observed in 49 patients (23.8%). HIF-1alpha protein overexpression was associated with significantly shorter overall and disease-free survival time (P = 0.003 and P = 0.001, respectively; Cox regression analysis). No correlation of HIF-1alpha and HER-2 expression or estrogen receptor density was observed. CONCLUSIONS: This study shows that HIF-1alpha is an independent prognostic factor for an unfavorable prognosis in patients with lymph node-positive breast cancer. Our results indicate that patients with advanced-stage breast cancers might profit from future therapies targeting HIF-1alpha.