[Lobular neoplasms and invasive lobular breast cancer]. / Lobuläre Neoplasie und invasives lobuläres Mammakarzinom.

The term lobular neoplasia (LN) comprises both atypical lobular hyperplasia (ALH), and lobular carcinoma in situ (LCIS) and thus a spectrum of morphologically heterogeneous but clinically and biologically related lesions. LN is regarded as a nonobligatory precursor lesion of invasive breast cancer and at the same time as an indicator lesion for ipsilateral and contralateral breast cancer risk of the patient. Rare pleomorphic or florid variants of LCIS must be differentiated from classical LCIS. The classical type of invasive lobular carcinoma (ILC) can be distinguished from the non-special type of invasive breast cancer (NST) by E-cadherin inactivation, loss of E-cadherin related cell adhesion and the subsequent discohesive growth pattern. Variant forms of ILC may show different molecular features, and solid and pleomorphic differentiation patterns in cases of high grade variants. Important parameters for the prognostic assessment of ILC are tumor grading and the recognition of morphological variants.

[Diagnostics of benign ductal epithelial cell proliferation of the breast in biopsy material]. / Diagnostik benigner duktaler Epithelproliferationen der Mamma in der Stanzbiopsie.

The pathological evaluation of radiological or sonographical abnormalities by needle core biopsy of the breast frequently involves the differential diagnosis of benign epithelial cell proliferations. The lesions to be considered include usual type and atypical ductal epithelial cell hyperplasia, columnar cell changes including flat epithelial cell atypia, the spectrum of hyperplastic and atypical apocrine epithelial cell proliferations and papillary lesions. This review provides an overview of the diagnostic criteria, the current terminology and the differential diagnosis of these lesions. The clinical management and the prognosis of the lesions are discussed.

[Ductal and lobular preneoplasia: role in breast cancer development]. / Duktale und lobuläre Präneoplasien: Biologische Bedeutung in der Entstehung invasiver Mammakarzinome.

BACKGROUND: With the widespread use of screening mammography and core needle biopsies, preneoplastic lesions of the breast are observed with increasing frequency. Unlike low-grade ductal carcinoma in situ (lg-DCIS), a possible precursor role of atypical ductal hyperplasia (ADH), lobular neoplasia (LN), and flat epithelial atypia (FEA) as well as their risk of progression have not been clearly defined. The aim of the present study was to evaluate the biological potential of these early low-grade lesions in the development of breast cancer. MATERIALS AND METHODS: Different series of precursor lesions and syn- or metachronous invasive carcinomas were tested for clonality using mitochondrial DNA sequencing, analysis of E-cadherin inactivation and comparative allelotyping with a panel of 14 polymorphous STR markers. RESULTS: In a series of invasive carcinomas occurring up to 10 years after the initial diagnosis of lobular neoplasia, a direct (clonal) association was established between LN and 3 of 5 invasive lobular cancers (ILC) but not in any of the cases with ductal secondary tumors. Tests on lg-DCIS and FEA occurring in association with tubular breast carcinomas demonstrated a direct clonal relationship in 67 and 70% of cases, respectively, and comparative allelotyping revealed a high degree of homology in the patterns of chromosomal imbalances. In addition, FEA and lg-DCIS, but not LN, were frequently closely related with each other. CONCLUSION: The present data provide molecular evidence for a direct clonal relationship of LN and ILC as well as of FEA and lg-DCIS with tubular breast carcinomas. However, the multifocal occurrence and frequent coexistence of the different precursor lesions suggests the presence of (possibly hormone-induced) field effects on the breast parenchyma.

Quality of life after emergency vs. elective esophagectomy with cervical reconstruction.

INTRODUCTION: Esophagectomy with reconstruction by collar anastomosis has an impact on the patients quality of life (QOL). The aim of this study was to explore a potential difference in QOL between elective and emergency esophagectomy with collar reconstruction. PATIENTS AND METHODS: Quality of life questionnaires were evaluated in 17 patients prior to esophagectomy, shortly after surgery, hospital discharge, and at least > 9 months after surgery using the EORTC QLQ C30 and EORTC OES 18 forms. In all patients reconstruction was per-formed by high collar anastomosis. Patients in group A received elective esophageal resection. In group B emergency esophagectomy was performed because of esophageal perforation for various reasons apart from cancer. In this group, delayed reconstruction was performed in a second operation 3-6 months after esophagectomy. RESULTS: There was a temporary decrease of postoperative QOL in both groups, which re-turned to preoperative values in the follow-up except for physical functioning, which remained decreased in group A (p < 0,05). There were no persisting differences in QOL after elective and emergency esophagectomy in the follow-up. DISCUSSION: Patients with elective and emergency esophagectomy and reconstruction by high collar anastomosis gained a good long-term QOL in our cohort of patients. This gives evidence that the observed QOL after elective resection of esophageal cancer is not only caused by a relief of cancer burden, but also due to a surgical procedure which is able to provide a good long-term QOL.

[Multiple space-occupying lesions of the liver: a rare differential diagnosis]. / Multiple Raumforderungen der Leber : Eine seltene Differenzialdiagnose.

The case of a 73-year-old female patient with a known increase in tumor markers is described. The course of this patient shows that a definitive diagnosis can sometimes not be achieved with comprehensive laboratory and imaging investigations alone.

[Villous adenoma within an urachal diverticulum associated with long-term mucusuria]. / Villöses Urachusdivertikeladenom mit Langzeitmukusurie.

During embryonic development the urachus connects the bladder with the umbilical cord. The urachus obliterates postnatally. Incomplete obliterations result in urachal cysts which, in the case of a connection to the bladder, are termed urachal diverticula. Urachal tumors are rare. We report on the case of a 46-year-old female patient with a 13-year history of recurrent bladder infections and mucusuria caused by an urachal diverticulum. After surgical resection histologic examination showed a mucus producing, intestinally differentiated adenoma within the diverticulum. This finding explains both the recurrent infections and the 10-year history of mucusuria.

[Clonal association of flat epithelial atypia and tubular breast cancer]. / Klonaler Zusammenhang flacher Epithelatypien und tubulärer Mammakarzinome.

Flat epithelial atypia (FEA) of the breast has recently gained attention as a possible precursor lesion of highly differentiated breast cancer. Especially tubular carcinomas, with which FEA shares cytological features, often occur in close proximity to each other. To examine a possible clonal relationship, we analysed mutations of the highly variable region of the mitochondrial genome in a series of tubular carcinomas, associated FEA and normal glands. Multiple sequence alignment showed identical mtDNA mutations in approximately 50% of paired FEA and tumour samples, indicative of a clonal relationship. Our data indicate a possible precursor role of FEA in the development of tubular breast cancer.

c-myc amplifications in primary breast carcinomas and their local recurrences.

OBJECTIVE: To evaluate the role of c-myc oncogene amplifications in the progression of invasive breast carcinomas. METHODS: c-myc gene copy number was evaluated in a series of 49 primary breast carcinomas and the corresponding local recurrences using fluorescence in situ hybridisation. RESULTS: 11 of the primary carcinomas (22%) harboured c-myc amplifications; these tumours typically were hormone receptor negative and occurred in younger patients (43 v 53 years). At the time of relapse, six additional tumours had acquired a c-myc amplification. The mean recurrence-free survival was 24 months; c-myc amplified tumours relapsed significantly earlier than carcinomas without amplification (18 v 27 months). Univariate analysis showed a worse overall survival in these patients. CONCLUSIONS: While c-myc amplifications can be observed in early stage breast cancer, especially in younger patients, they often occur later in tumour development and appear to be associated with disease progression.

Molecular characterisation of pancreatic ductal adenocarcinoma in patients under 40.

BACKGROUND: Pancreatic ductal adenocarcinoma (PDAC) rarely affects people under 40. OBJECTIVES: To determine whether the clinical, pathomorphological and genetic features of PDAC occurring in young patients (

Apoptosis resistance in pigmented villonodular synovitis.

OBJECTIVE: Pigmented villonodular synovitis (PVNS) is a proliferative lesion originating from synovial tissue with a locally aggressive behaviour. We analysed the pathogenetic role of apoptosis resistance for sustained cell proliferation in PVNS. METHODS: The expression of bcl-2, p53 and Ki-67 was examined in 80 cases of PVNS using immunohistochemistry. In 43 of these cases, DNA content and distribution of cell-cycle phases were investigated by flow cytometry. Additionally, 10 cases of PVNS were analysed by multi-parametric flow cytometry for expression of p53, caspase3, and bcl-2 and by TUNEL to detect DNA fragmentation. RESULTS: No apoptotic cell fractions were detected in any investigated cases. Expression of bcl-2 was found in 84% of cases (up to 6.5% of cells) and was significantly associated with DNA-fragmentation observed by TUNEL (p=0.037). Orthologous p53 expression was observed in 37% of cases. The level of p53 expression correlated with the proliferative activity and the expression of both caspase3 (p=0.017) and bcl-2 (p=0.0013). (No statistically significant correlations between expression of bcl-2, p53, caspase3, DNA fragmentation or proliferative index and age, sex of patients, disease recurrence, growth pattern or size of lesion were found). CONCLUSION: Apoptosis resistance is a critical event in the progression of PVNS and may contribute to the survival of the proliferating synovial cells in PVNS and to the permanent slow progression of these lesions.

The location of KIT and PDGFRA gene mutations in gastrointestinal stromal tumours is site and phenotype associated.

AIMS: To assess the relation between KIT and PDGFRA mutations and the site of origin, histological phenotype, and pathomorphologically determined risk assessment in gastrointestinal stromal tumours (GISTs). METHODS: A series of 83 clinicopathologically characterised GISTs from 79 patients was analysed for KIT and PDGFRA mutations by polymerase chain reaction amplification, single strand conformation polymorphism analysis, and direct DNA sequencing. RESULTS: KIT or PDGFRA mutations were found in 57 and 11 GISTs, respectively. Most KIT mutations involved exon 11 (46 cases), followed by exon 9 (10 cases). The PDGFRA mutations mostly affected exon 18 (eight cases), followed by exon 12 (three cases). There was a significant association between KIT exon 9 mutations and an intestinal origin of GISTs, and between PDGFRA mutations and gastric origin of the tumours. In addition, the presence of PDGFRA mutations was significantly associated with epithelioid/mixed histology, as was the absence of identified receptor tyrosine kinase mutations. Vice versa, KIT exon 11 mutations were almost exclusively found in spindle cell GISTs. Furthermore, the presence of any KIT and PDGFRA mutations and the presence of KIT mutations alone were significantly associated with high risk/malignant GISTs. CONCLUSIONS: The location of KIT and PDGFRA mutations in GISTs is associated with the site of origin and histological phenotype. Genotyping of GISTs may be a helpful additional parameter in determining the biological profile of these tumours.

CT-guided irreversible electroporation in an acute porcine liver model: effect of previous transarterial iodized oil tissue marking on technical parameters, 3D computed tomographic rendering of the electroporation zone, and histopathology.

PURPOSE: To evaluate the effect of previous transarterial iodized oil tissue marking (ITM) on technical parameters, three-dimensional (3D) computed tomographic (CT) rendering of the electroporation zone, and histopathology after CT-guided irreversible electroporation (IRE) in an acute porcine liver model as a potential strategy to improve IRE performance. METHODS: After Ethics Committee approval was obtained, in five landrace pigs, two IREs of the right and left liver (RL and LL) were performed under CT guidance with identical electroporation parameters. Before IRE, transarterial marking of the LL was performed with iodized oil. Nonenhanced and contrast-enhanced CT examinations followed. One hour after IRE, animals were killed and livers collected. Mean resulting voltage and amperage during IRE were assessed. For 3D CT rendering of the electroporation zone, parameters for size and shape were analyzed. Quantitative data were compared by the Mann-Whitney test. Histopathological differences were assessed. RESULTS: Mean resulting voltage and amperage were 2,545.3 ± 66.0 V and 26.1 ± 1.8 A for RL, and 2,537.3 ± 69.0 V and 27.7 ± 1.8 A for LL without significant differences. Short axis, volume, and sphericity index were 16.5 ± 4.4 mm, 8.6 ± 3.2 cm(3), and 1.7 ± 0.3 for RL, and 18.2 ± 3.4 mm, 9.8 ± 3.8 cm(3), and 1.7 ± 0.3 for LL without significant differences. For RL and LL, the electroporation zone consisted of severely widened hepatic sinusoids containing erythrocytes and showed homogeneous apoptosis. For LL, iodized oil could be detected in the center and at the rim of the electroporation zone. CONCLUSION: There is no adverse effect of previous ITM on technical parameters, 3D CT rendering of the electroporation zone, and histopathology after CT-guided IRE of the liver.

Multigene Assays for Classification, Prognosis, and Prediction in Breast Cancer: a Critical Review on the Background and Clinical Utility.

Gene signatures which are based on multigene profiling assays have been developed for the purpose to better define the prognosis and prediction of therapy results in early-stage breast cancer. These assays were designed to be more specific than conventional clinico-pathologic parameters in the selection of patients for (neo-)adjuvant treatment and in effect help to avoid unnecessary cytotoxic treatment. In this review we describe molecular risk scores, for which tests are commercially available (PAM50®, MammaTyper®, MammaPrint®, Oncotype DX®, Endopredict®, Genomic Grade Index®) and IHC risk scores (Mammostrat® and IHC4), and discuss the current evidence of their clinical use.

Strong EGFR signaling in cell line models of ERBB2-amplified breast cancer attenuates response towards ERBB2-targeting drugs.

Increasing the efficacy of targeted cancer therapies requires the identification of robust biomarkers suitable for patient stratification. This study focused on the identification of molecular mechanisms causing resistance against the anti-ERBB2-directed therapeutic antibodies trastuzumab and pertuzumab presently used to treat patients with ERBB2-amplified breast cancer. Immunohistochemistry and clinical data were evaluated and yielded evidence for the existence of ERBB2-amplified breast cancer with high-level epidermal growth-factor receptor (EGFR) expression as a separate tumor entity. Because the proto-oncogene EGFR tightly interacts with ERBB2 on the protein level, the hypothesis that high-level EGFR expression might contribute to resistance against ERBB2-directed therapies was experimentally validated. SKBR3 and HCC1954 cells were chosen as model systems of EGFR-high/ERBB2-amplified breast cancer and exposed to trastuzumab, pertuzumab and erlotinib, respectively, and in combination. Drug impact was quantified in cell viability assays and on the proteomic level using reverse-phase protein arrays. Phosphoprotein dynamics revealed a significant downregulation of AKT signaling after exposure to trastuzumab, pertuzumab or a coapplication of both antibodies in SKBR3 cells but no concomitant impact on ERK1/2, RB or RPS6 phosphorylation. On the other hand, signaling was fully downregulated in SKBR3 cells after coinhibition of EGFR and ERBB2. Inhibitory effects in HCC1954 cells were driven by erlotinib alone, and a significant upregulation of RPS6 and RB phosphorylation was observed after coincubation with pertuzumab and trastuzumab. In summary, proteomic data suggest that high-level expression of EGFR in ERBB2-amplified breast cancer cells attenuates the effect of anti-ERBB2-directed antibodies. In conclusion, EGFR expression may serve as diagnostic and predictive biomarker to advance personalized treatment concepts of patients with ERBB2-amplified breast cancer.

Outcome analysis of patients with primary breast cancer initially treated at a certified academic breast unit.

INTRODUCTION: Evaluation of oncological outcome and prognostic factors of patients with primary breast cancer treated at a certified academic breast unit. PATIENTS AND METHODS: We prospectively collected data of 3338 patients, diagnosed with primary breast cancer between 01.01.2003 and 31.12.2010 and treated at the Breast Unit Heidelberg, Germany, in order to analyze outcome in clinical practice. We evaluated local control rate (LCR), disease-free survival (DFS), distant disease-free survival (DDFS), observed overall survival (OS) and age-adjusted relative overall survival (ROS). In addition, the impact of known prognostic factors on these outcome variables was examined in univariate and multivariate analyses. RESULTS: Of all patients, 368 (11.0%) had carcinoma in situ (CIS) and 197 (5.9%) had bilateral cancers. For the 2970 patients with invasive cancer, of which 49 patients (1.7%) had metastastic disease at time of diagnosis, DFS, LCR, DDFS, OS and ROS at 5 years were 79.8%, 84.7%, 81.2%, 86.3%, and 89.8%, respectively. In multivariate analysis age, pT category, nodal status, hormone receptor status and grading were identified as independent prognostic factors for OS. CONCLUSION: Compared with recent population-based reports from Germany, more favourable patient characteristics and nominally higher survival was found among this large cohort of patients with primary breast cancer treated at a single certified breast unit.

Deubiquitination of EGFR by Cezanne-1 contributes to cancer progression.

Once stimulated, the epidermal growth factor receptor (EGFR) undergoes self-phosphorylation, which, on the one hand, instigates signaling cascades, and on the other hand, recruits CBL ubiquitin ligases, which mark EGFRs for degradation. Using RNA interference screens, we identified a deubiquitinating enzyme, Cezanne-1, that opposes receptor degradation and enhances EGFR signaling. These functions require the catalytic- and ubiquitin-binding domains of Cezanne-1, and they involve physical interactions and transphosphorylation of Cezanne-1 by EGFR. In line with the ability of Cezanne-1 to augment EGF-induced growth and migration signals, the enzyme is overexpressed in breast cancer. Congruently, the corresponding gene is amplified in approximately one third of mammary tumors, and high transcript levels predict an aggressive disease course. In conclusion, deubiquitination by Cezanne-1 curtails degradation of growth factor receptors, thereby promotes oncogenic growth signals.

Gains of chromosome region 3q26 in intraepithelial neoplasia and invasive squamous cell carcinoma of the vulva are frequent and independent of HPV status.

AIMS: Two different forms of vulvar intraepithelial neoplasia (VIN) are recognised: (1) usual-type (bowenoid) VIN, which is related to high-risk papillomavirus infection, and (2) differentiated (simplex) VIN, which is associated with chronic inflammation. The aim of this study was to investigate the presence of chromosome 3q26 gains in the spectrum of precancerous lesions and invasive squamous cell carcinomas (SCCs) of the vulva. METHODS: 3q26 gains were analysed using fluorescence in situ hybridisation in a series of usual-type VINs, VINs of the differentiated type and invasive squamous cell carcinomas. In addition, all cases were examined for human papillomavirus (HPV) DNA, p53 mutations, and p16 and p53 protein expression. RESULTS: Gains of chromosome 3q26 were present in all VINs of the differentiated type and in 50% of the usual-type VIN lesions. 81% of SCCs were positive for 3q26 gains irrespective of the HPV status and of the associated precursor lesion. HPV-associated lesions exhibited the typical, strong cytoplasmic p16 accumulation while mutated p53 was only detected in HPV-negative VINs or SCCs, and was associated with an overexpression of p53 protein. CONCLUSIONS: Immunohistochemical evaluation of p16 and p53 expression aids in the differential diagnosis of squamous cell alterations of the vulva. However, detection of 3q26 imbalance is of additional diagnostic value in difficult cases of HPV-unrelated usual-type VINs and VINs of the differentiated type.

Detection of the ASPSCR1-TFE3 gene fusion in paraffin-embedded alveolar soft part sarcomas.

AIMS: Alveolar soft part sarcoma (ASPS) is a rare soft tissue tumour with unique morphology and a recurrent, non-reciprocal translocation der(17)t(X;17)(p11.2;q25) leading to the fusion of ASPSCR1 (also known as ASPL) to the transcription factor TFE3. Although diagnosis is straightforward in classical cases, tumours with atypical morphological features may be difficult to classify solely on the basis of conventional histopathology. The aim of this study was to analyse the chromosomal breakpoints in paraffin-embedded tissue. METHODS AND RESULTS: Three male and two female ASPS patients including one case with uncommon histology were investigated by fluorescence in situ hybridization with split- and fusion-probes. The presence of the resulting ASPSCR1-TFE3 fusion transcripts was assessed by reverse transcriptase-polymerase chain reaction. Hybridization results showed a t(X;17)(p11.2;q25) in all tumours with a duplication of the telomeric part of chromosome Xp. In addition to wild-type TFE3, ASPSCR1-TFE3 fusion transcripts (three type 1 and two type 2 transcripts) were detected in all cases. CONCLUSIONS: Molecular confirmation of ASPSCR1-TFE3 gene fusion is applicable to routinely processed archival and diagnostic tumour samples and aids in the differential diagnosis of ASPS.

[Lobular carcinoma in situ (LCIS): risk factor and precursor of invasive lobular breast cancer]. / Lobuläres Carcinoma in situ (LCIS): Risikofaktor und Vorläufer invasiver lobulärer Mammakarzinome.

Lobular carcinoma in situ (LCIS) of the breast typically is an incidental finding in breast biopsies performed for a variety of reasons. As patients with LCIS have a bilaterally increased risk of developing invasive ductal or lobular breast cancer, the lesion is presently considered an indicator rather than a direct precursor of breast cancer. However, this view is challenged by the finding that LCIS often accompanies invasive lobular carcinomas (ILC) and that the frequency of ILC following LCIS is far higher compared to unselected patients. To further examine the role of LCIS in the development of invasive breast cancer, we analysed a series of patients with pure LCIS, who later developed invasive breast cancer. Mitochondrial D-loop sequencing revealed a clonal relationship of LCIS and a subset of ILC occurring in the same breast between 2 and 10 years later. Apparently, LCIS has a chimeric role in breast cancer development and is not only a risk factor but in some cases a precursor of ILC.

[Concepts and problems of lobular neoplasia]. / Konzept und Problematik der lobulären Neoplasie.

The term lobular neoplasia (LN) includes lobular carcinoma in situ (LCIS) and atypical lobular neoplasia (ALH). It is generally considered to be a risk lesion and a non-obligatory precursor for the subsequent development of an invasive carcinoma in the ipsilateral or contralateral breast. LN has also been termed lobular intraepithelial neoplasia (LIN). A grading system (LIN 1-LIN 3) has been suggested as a tool for a more precise estimation of the individual risk. When LN is the most significant finding in a core biopsy, the probability of a higher grade lesion is about 17% in the follow-up surgical biopsy, justifying follow-up surgery in the majority of cases. A higher risk of progression is attributed to LIN 3 (pleomorphic LN, extensive LN, and signet ring cell LN) compared to LIN 1 or LIN 2. These special forms of LN may have an unusual presentation clinically or histologically. Using immunohistology, LN are characterized by the loss of E-cadherin, low proliferative activity and by positive hormone receptor status. The molecular characteristics of LN are similar to those of invasive lobular carcinomas, indicating the nature of LN as a precursor lesion.