To address the value of qRT-PCR and IHC in accurately detecting lymph node micrometastasis in gynecological cancer, we performed a systematic approach, using a set of dual molecular tumor-specific markers such as cytokeratin 19 (CK19) and carbonic anhydrase 9 (CA9), in a series of 46 patients (19 with cervical cancer, 18 with endometrial cancer, and 9 with vulvar cancer). A total of 1281 lymph nodes were analyzed and 28 were found positive by histopathology. Following this documentation, 82 lymph nodes, 11 positive and 71 negative, were randomly selected and further analyzed both by IHC and qRT-PCR for CK19 and CA9 expression. All 11 (100%) expressed CK19 by IHC, while only 6 (54.5%) expressed CA9. On the contrary, all the histologically negative for micrometastases lymph nodes were also negative by IHC analysis for both markers. The comparative diagnostic efficacy of the two markers using qRT-PCR, however, disclosed that the analysis of the same aliquots of the 82 lymph nodes led to 100% specificity for the CK19 biomarker, while, in contrast, CA9 failed to recapitulate a similar pattern. These data suggest that qRT-PCR exhibits a better diagnostic accuracy compared to IHC, while CK19 displays a consistent pattern of detection compared to CA9.

The molecular mechanisms of vulvar squamous cell carcinoma (VSCC) remain obscure. To this end, we investigated systematically for the first time the expression profile of VSCC using the microarray technology, in a total of 11 snap-frozen samples, from five VSCC patients covering early and advanced stages of VSCC undergoing radical surgery and from six matched healthy controls. All experiments were performed using Affymetrix Human Genome U133A 2.0 oligonucleotide arrays, covering 22,277 probe sets. Genes were filtered and analyzed using analysis of variance, t test, fold-change calculations, and unsupervised hierarchical cluster analysis. Further processing included functional analysis and overrepresentation calculations based on Gene Ontology, Database for Annotation, Visualization, and Integrated Discovery, and Ingenuity Pathway Analysis. The molecular phenotypes of VSCC patients exhibited significant and discrete transcriptional differences from the healthy controls, whereas principal component analysis documented that this separation is mediated by a consistent set of gene expression differences. We detected 1077 genes (306 upregulated and 771 downregulated) that were differentially expressed between VSCC patients and healthy controls by at least twofold (P < .01), whereas a novel subset of patients was revealed displaying a distinct pattern of 125 upregulated genes involved in multiple cellular processes. Functional analysis of the 1077 genes documented their involvement in more than 50 signaling pathways, such as PTEN, oncostatin M, and extracellular signal-regulated kinase signaling, affecting extracellular matrix remodeling and invasion. Comparison of our data set with those of the single VIN study revealed that the two entities share a limited number of genes and display unique features.

OBJECTIVE:The rarity of uterine sarcomas along with their pathological and molecular heterogeneities render their study particularly challenging. We evaluated a panel of somatic mutations principally centering on the tyrosine kinase gene family and their downstream signaling cascades in an attempt to identify potential candidate markers that may assist in diagnostic or therapeutic decisions in these tumors. METHODS:We performed mutational analysis of 20 exons from 9 genes (EGFR, CDKN2A, MET, KIT, RAS, BRAF, PI3KCA, HER-2 and PDGFR-alpha) on biopsy material from 25 patients who underwent primary surgery for uterine sarcoma between October 1995 and October 2003. Due to the limited number of studies conducted we have also undertaken a literature review of somatic mutations in uterine sarcomas. RESULTS:A total of 3 different somatic mutations were identified: one KRAS (codon G12D) in a carcinosarcoma and two exon 20 PI3KCA mutations (H1047R and H1047Y) both in carcinosarcomas. Mutational status of all mutations was confirmed using germline DNA extracted from peripheral blood. Consistent with the literature data, no other mutations regarding the rest of the genes of the panel were identified. Due to the low number of somatic mutations in our series, we did not perform further clinicopathological correlations. CONCLUSION:The absence of somatic mutations in the majority of genes that are considered critical in neoplastic transformation hampers the identification of potential therapeutic targets in patients with uterine sarcoma.

Purpose: Colposcopy occupies a key role in the prevention of cervical cancer by identifying preinvasive or invasive lesions. However, colposcopy is subjective and is responsible for 52% of screening failures. Dynamic spectral imaging (DSI) is based on the objective, quantitative assessment of the acetowhitening effect. This study compared DSI with colposcopy. Experimental Design: Women referred for colposcopy were examined simultaneously with colposcopy and DSI using a precommercial DySIS model (FPC-03) in an international, multicenter trial. The colposcopy impression and DySIS values were compared with consensus histology reports of biopsies. Subjects were recruited to a training group and subsequently to a test group. Measures were taken to avoid verification bias. Results: The training and test groups comprised 82 and 308 eligible women, respectively. A cutoff value to identify high-grade disease was selected from the results of the training group and data from previous work. Receiver operator curve analysis of the test data showed an area under the curve of 0.844. DySIS detected 62.9% more high-grade cases than colposcopy (57 versus 35, P = 0.0001). DySIS exceeded end points approved by the Food and Drug Administration for similar studies, with increments in the true positive rate of 22/308 (7.1%; lower 95% CL, 4.5% versus 2%) and in the false positive rate of 32/308 (10.4%; upper 95% CL, 14.7% versus 15%). Conclusions: DySIS is more sensitive than colposcopy in detecting high-grade lesions and can provide improved guidance for biopsy. The results are obtained in a user-independent fashion, making it suitable for use by nursing personnel.

PURPOSE OF INVESTIGATION Primary fallopian tube carcinoma (PFTC) is a rare malignancy with only few data existing on the impact of prognostic factors. METHODS We retrospectively analyzed 26 patients. Tissue blocks were reviewed and sections were stained for vascular endothelial growth factor (VEGF), matrix metalloproteinases 2 and 9 (MMP-2, MMP-9), tissue inhibitors of metalloproteinases 1 and 2 (TIMP-1, TIMP-2), c-erbB-2, estrogen (ER), and progesterone receptors (PgR). RESULTS Reactivity for VEGF, ER, PgR, MMP-2, MMP-9, TIMP-1, TIMP-2 and c-erbB-2 was observed in 85%, 46%, 27%, 11.5%, 58%, 0%, 23% and 8% of specimens, respectively. None of the markers studied displayed prognostic significance. Regarding clinical prognostic factors, the hazard ratio (HR) for progression and death for patients with tumor residuum > 2 cm was 5.24 (p < 0.01) and 11.19 (p < 0.005), respectively. Patients with advanced stage disease had a HR of 12.55 (p < 0.05) for progression, while the HR for death was not found to be statistically significant. CONCLUSION None of the biomarkers studied seems to influence survival. Early-stage disease and optimal debulking are associated with improved outcome.

BACKGROUND:Akt is a serine/threonine kinase which is fully activated when phosphorylated (pAkt). The aim of this study was to investigate the expression pattern of phosphorylated Akt at Threonine 308 [pAkt(Thr308)] in association with clinicopathological parameters and various biological markers.MATERIALS AND METHODS:Immunohistochemistry was performed on paraffin-embedded tissue specimens from 152 invasive breast carcinomas to detect the expression of the proteins pAkt(Thr308), estrogen (ER) and progesterone (PR) receptors, p53, Ki-67 and c-erbB-2.RESULTS:pAkt(Thr308) protein was immunodetected in the cytoplasm and the nuclei of the malignant cells. pAkt was found to be positively associated with the lobular histological type, while it was found to exert no impact on patients' survival. pAkt(Thr308) immunopositivity was inversely related to Ki-67 and p53 (p=0.013 and p=0.020, respectively), while being positively associated with cerbB2 expression (p=0.005).CONCLUSION:This is the first study to show a frequent detection of pAkt(Thr308) in lobular breast carcinomas and an association of its expression with indices of proliferation (c-erbB2, Ki-67) and apoptosis (p53).

Objectives. Vascular endothelial growth factors A and B (VEGF-A and VEGF-B) play a major role in angiogenesis and activate VEGF receptor 1 (VEGFR-1). However, the clinicopathologic and clinical value of VEGF-B and VEGFR-1 in invasive breast carcinoma remains unclear. Methods. We immunohistochemically examined the expression pattern of VEGF-A, VEGF-B and VEGFR-1 in 177 invasive breast carcinomas in relation to clinicopathological parameters, p53, c-erbB2 proteins expression and patients' survival. Results. VEGF-A, VEGF-B and VEGFR-1 were immunodetected predominantly in the cytoplasm of the malignant cells. None of the studied markers correlated with any of the clinicopathological parameters, other than stromal VEGFR-1 which inversely correlated with PR (p=0.021). Cancerous VEGF-A and stromal VEGFR-1 were positively related to p53 (p=0.016 and p=0.033, respectively). Cancerous VEGF-B was positively associated with c-erbB-2 (p=0.045) and was found to exert an unfavorable impact on both disease-free and the overall survival of the node-positive patients (p=0.05 and p=0.029, respectively). Cancerous VEGFR-1 was recognized as being an independent poor prognostic indicator (p=0.037). Conclusion. These findings suggest that, while VEGF-B seems to be useful as a prognostic indicator only in node-positive patients, VEGFR-1 may be an independent poor prognosticator in patients with invasive breast carcinoma.

Introduction: Splenosis is the autoimplantation of ectopic spleen tissue in various anatomic cavities of the body resulting after trauma or rupture of the splenic parenchyma. The major localization sites of this phenomenon are mainly intraperitoneal, the gastrenteric tract, genitalia, intrahepatically and the kidneys. Extraperitoneal locations occur less frequently and include the thorax and brain. Also locallization in the subcutaneous fat has been described. Case report: We present the case of a 32-year-old woman with symptomatic peritoneal cavity splenosis occurring ten years after traumatic splenectomy. The patient was admitted to our department with the clinical presentation of an adnexal tumor. US and CT confirmed an adnexal mass. Exploratory laparotomy was performed and multiple focal lesions were noticed on the uterus, ovaries and intestinal tract. Biopsies were taken and sent for histological analysis. The pathology specimen revealed ectopic splenic tissue. After surgical intervention the patient remained asymptomatic. Conclusion: Splenosis is a rare phenomenon which clinicians should be aware of in order to spare patients from pointless surgical interventions. Patients with abdominal masses and post-traumatic splenectomy should be checked for splenosis.

Einführung: Grund für diese Studie war die Messung der Expression der Onkogene C-myc, HER-2/neu und das extra zelluläre Matrixglukoprotein Tenascin in Uterussarkomen in Bezug auf den klinischen Verlauf (Überlebens- und Rezidiv-Raten) zu untersuchen. Patienten und Methoden: 36 Patientinnen mit Uterussarkomen nahmen an der Studie teil, wobei 22 an einen Müller'schen Mischtumor, 10 an Leiomysarkom und 4 an anderen Sarkomarten erkrankt waren. C-myc, Tenascin und HER-2/neu Expression wurde in allen 36 Fällen mittels immunhistochemischer Methoden bestimmt. Ergebnisse: C-myc war in 23 von 36 Fällen (64%), Tenascin in 15 von 36 Fällen (42%) und HER-2/neu in 8 von 36 Fällen (22%) positiv. Überlebens- und Rezidivraten unterschieden sich nicht signifikant bezüglich positiver und negativer Expression bei allen 3 getesteten Substraten in dieser Studie. Schlussfolgerung: Bei den 36 Uterussarkomen dieser Studie (22 Müller'sche Mischtumore, 10 Leiomysarkome und 4 andere Sarkome) war die untersuchte Expression von 3 Proteinen nicht mit einer erhöhten oder verminderten Überlebens- oder Redzidiv-Rate assoziiert.

Einführung: Die Aufgabe dieser prospektiven Studie war es nach Anwendung eines spezifischen therapeutischen Protokolls die Diagnose (Überlebensrate, Rezidivrate) zu beurteilen, sowie prognostischen und epidemiologische Faktoren im Hinblick auf uterine Sarkome. Patienten und Methode: Das Protokoll bestand aus einer radikalen Operation zur Zytoreduktion und kombiniert mit einer adjuvanten Chemotherapie mit verschiedenen Chemotherapeutika (Iphosphamid, Etoposid, Doxyrubicin, MESNA). Die Studie bestand aus 36 Patientinnen ohne Rückbildungs- oder Progressionstendenz nach der Chirurgie (23 Fälle = 61 % waren Müllersche Mischtumoren, 10 Fälle = 28 % waren Leiomyosarkome und 4 = 11 % waren andere Arten). Ergebnisse: Das durchschnittliche Alter bei der Diagnose war 63,5 Jahre für die Müllerschen Mischtumore, 52,8 Jahre für die Leiomyosarkome und 38,7 Jahre für die anderen Typen. Hierbei handelt es sich um eine signifikante Altersdifferenz (p=0,008).Zusätzlich war der Anteil von prämenopausalen Patientinnen signifikant bei den Tumoren mit anderen histologischen Arten erhöht. Nach einer medianen Nachkontrollzeit von 61,5 Jahren fand sich eine mediane Überlebenszeit von 46,8 Monaten und die mediane krankheitsfreie Periode betrug 29,5 Monate im Gesamtkollektiv. Der Verlauf wurde als klinisch zufrieden stellend bewertet, wenn man in Betracht zieht, dass diese Tumoren aggressiv sind. Für Müllersche Mischtumoren lag die mediane Überlebenszeit bei 46,8 Monaten und bei den Leiomyosarkomen betrug sie 44,1 Monate. Eine Differenz, die nicht signifikant war. Im Gegensatz dazu traten bei Patientinnen mit Leimysarkommen schneller Rezidive auf als bei denen mit Müllerschen Mischtumoren oder anderen Typen. Die Differenz war signifikant (p=0,01). Bei den Sarkomen scheint das Leiomyosarkom die aggressivste histologische Form zu sein. Das Stadium bei der Diagnose (früh 1 und 2 gegenüber fortgeschritten 3 und 4) war nicht signifikant unterschiedlich bezogen auf Überlebenszeit und Rezidivrate. Dies war durch die relativ hohe Überlebenszeit und krankheitsfreie Periode bedingt, die in den fortgeschrittenen Stadien erzielt wurde, wahrscheinlich aufgrund einer erfolgreichen Chemotherapie nach der radikalen Operation. Bei den Müllerschen Mischtumoren war Gefäßinvasion und das Vorhandensein von heterologen Elementen nicht entscheidend für den klinischen Verlauf obzwar sich im letzteren Falle eine Tendenz zur schlechteren Prognose ablesen ließ. Schlussfolgerung: Trotz der Tumoraggressivität und der niedrigen Überlebenszeit und krankheitsfreien Periode scheint ein komplexes therapeutisches Protokoll mit radikaler operativer Zytoreduktion und adjuvanter postoperativer Chemotherapie mit verschiedenen Kombinationen zu höheren Überlebensraten und längerer Rezidivfreiheit beizutragen. Die adjuvante Radiotherapie scheint den klinischen Verlauf nicht zu beeinflussen. Die Leiomyosarkome erscheinen als die aggressivste histologische Form innerhalb der im weiblichen Genitaltrakt auftretenden Sarkome, da sie sehr rasch rezidivieren.

Smad2 participates in the TGF-β signaling pathway, where it cooperates with transcription factors to regulate expression of defined genes. The purpose of this study was to investigate the expression pattern of phosphorylated Smad2 (pSmad2) in association with clinicopathological parameters and biological markers of proliferation and invasion. Immunohistochemistry was applied on paraffin-embedded sections from 164 patients with invasive breast carcinomas to detect the expression of the proteins pSmad2, ER, PR, Ki67, topoisomerase IIa, ERK2, catenin-p120, MMP-14 and TIMP-2. pSmad2 protein was detected in the nuclei of the malignant cells (68.1%) and in the tumor fibroblasts (55.2%). Nuclear pSmad2 was inversely correlated with histological grade and LN (p=0.047 and p=0.05) as well as with Ki67 and topoIIa (p=0.003 and p=0.021, respectively). There was also an inverse relation between nuclear pSmad2 and normal immunoexpression of the adhesion molecule catenin-p120 (p=0.028). Both nuclear and stromal pSmad2 were positively correlated with ERK2 of tumor fibroblasts (p=0.008 and p=0.0001, respectively), while stromal pSmad2 was furthermore related to stromal MMP-14 and tumor TIMP-2 (p=0.006 and p=0.022, respectively). Patients with high expression of cancerous pSmad2 tended to have a better prognosis, although statistic significance was never reached. pSmad2 was found to play a dual role, according to its distribution. Nuclear localization was thus found to be related to a less aggressive tumor phenotype, whereas stromal location was associated with an invasive phenotype.

PURPOSE OF INVESTIGATION Uterine sarcomas are rare neoplasms characterized by a high rate of local recurrences and distant metastases. The role of chemotherapy in early-stage completely resected disease remains controversial. METHODS Thirty-one patients with Stage I or II uterine sarcomas, referred to our center for adjuvant chemotherapy, received anthracycline-based regimens. Seventeen (54.8%) patients received ifosfamide, etoposide and epirubicin, six (19.4%) were treated with doxorubicin and carboplatin, three (9.6%) were administered doxorubicin and ifosfamide, while five (16.1%) patients received various anthracycline-based regimens. RESULTS With a median follow-up of 82 months disease recurred in 12 (38.7%) patients. Five-year survival probability is estimated at 54%. Both median overall survival and time to progression for all patients have not been reached yet. Patients who received ifosfamide-containing regimens had a statistically significant benefit in overall survival (p < or = 0.05) when compared with those treated with non-ifosfamide-containing regimens. CONCLUSION Our data suggest a potential role for anthracycline- and ifosfamide-containing chemotherapy in the adjuvant setting for early-stage uterine sarcomas.