Virchows Archiv

Morra, Laura; Moch, Holger

doi: 10.1007/s00428-011-1151-5pmid: 21997759

Periostin, also called osteoblast-specific factor 2, is a secreted cell adhesion protein, which shares a homology with the insect cell adhesion molecule fasciclin I. It has been shown to be an important regulator of bone and tooth formation and maintenance, and of cardiac development and healing. Recent studies revealed that periostin plays an important role in tumor development and is upregulated in a wide variety of cancers such as colon, pancreatic, ovarian, breast, head and neck, thyroid, and gastric cancer as well as in neuroblastoma. Periostin binding to the integrins activates the Akt/PKB- and FAK-mediated signaling pathways which lead to increased cell survival, angiogenesis, invasion, metastasis, and importantly, epithelial-mesenchymal transition of carcinoma cells. In this review we summarize recent clinicopathological studies that have investigated periostin expression in lung, kidney, prostate, liver cancer, and malignant pleural mesothelioma and discuss the role of periostin isoforms in tumorigenesis and their potential as targets for stroma-targeted anticancer therapy.

Catta-Preta, Mariana; Mendonca, Leonardo; Fraulob-Aquino, Julio; Aguila, Marcia; Mandarim-de-Lacerda, Carlos

doi: 10.1007/s00428-011-1147-1pmid: 21901430

The issue of adequately quantitatively evaluating hepatic steatosis is still unresolved. Therefore, we compared three methods of quantitative assessment. Two groups of mice (n = 10 each) were fed standard chow (10% fat, SC group) or a high-fat diet (60% fat, HF group) for 16 weeks, and hepatic triglyceride (HT) and liver tissue were then studied. Paraplast-embedded tissues stained by hematoxylin and eosin (H-E) were compared to frozen sections stained by Oil Red-O (ORO). In addition, the volume density of steatosis (Vv[steatosis, liver]) was measured by point counting (P-C, sections H-E or ORO) or by image analysis (I-A, sections ORO). HT was significantly higher in the HF group (104% greater, P = 0.0004) than in the SC group. With P-C and H-E, Vv[steatosis, liver] was 4.80 ± 0.90% in the SC group and 33.50 ± 3.17% in the HF group (600% greater, P < 0.0001). With P-C and ORO, Vv[steatosis, liver] was 4.86 ± 0.89% in the SC group and 25.21 ± 1.27% in the HF group (420% greater, P < 0.0001). With I-A and ORO, Vv[steatosis, liver] was 4.17 ± 0.85% in the SC group and 23.35 ± 1.58% in the HF group (460% greater, P < 0.0001). Correlations between Vv[steatosis, liver] and HT were strong and significant in all methods. In conclusion, all methods were appropriate and reproducible. In P-C and H-E, there is a slight overestimation of steatosis in the HF animals in comparison to frozen sections and ORO; in frozen sections, differences between P-C and I-A are insignificant.

Farber, Liora; Efrati, Edna; Elkin, Hela; Peerless, Yehudit; Sabo, Edmond; Ben-Izhak, Ofer; Hershkovitz, Dov

doi: 10.1007/s00428-011-1158-ypmid: 22016105

KRAS mutation status has a significant role determining anti-epidermal growth factor receptor (anti-EGFR) treatment response in colon carcinoma patients. Malignant transformation is a dynamic process and therefore, it is conceivable that, at a certain point, the tumor cells’ mass might be heterogeneous for particular mutations. Therefore, the fraction of tumor cells carrying a particular mutation may be more relevant for treatment than the simple determination of presence or absence of mutation. The purpose of this study is to assess whether or not KRAS mutation status is heterogeneous and, if so, to what extent in colon carcinoma samples. Deoxyribonucleic acid was extracted from formalin-fixed paraffin-embedded samples of colon carcinoma and analyzed for the presence of KRAS mutation. The relative fraction of mutated versus wild-type KRAS alleles was evaluated by real-time polymerase chain reaction. Additionally, the relative fraction of cancer cells in the tissue sample was evaluated using computer assisted morphometric analysis. Using this data, we calculated the fraction of mutation containing cells in the samples. Colon carcinoma (169 cases) were analyzed, and a KRAS mutation was found in 75 cases (44%), of which 42 were available for morphometric analysis. In 41 (97.6%) of these cases, the fraction of mutation containing tumor cells was 50% or higher, indicating the absence of significant KRAS mutation status heterogeneity. There was a strong positive correlation (R = 0.66, P < 0.0001) between the fraction of mutated KRAS alleles and the fraction of cancer cells in the samples. The strong positive correlation between the fraction of mutated KRAS alleles and the fraction of cancer cells in the samples indicate homogeneity of KRAS mutation status in colorectal carcinoma.

Choi, Song-Yi; Kim, Seok; Kim, Won; Kim, Kihyun; Ko, Young-Hyeh

doi: 10.1007/s00428-011-1153-3pmid: 22002677

Aggressive B cell lymphomas of the gastrointestinal tract include Burkitt lymphoma (BL), diffuse large B cell lymphoma, and B cell lymphoma, unclassifiable, with features intermediate between DLBCL and BL. To investigate the clinical characters of DLBCL/BL of the gastrointestinal (GI) tract, 101 cases of aggressive B cell lymphoma of the GI tract were analyzed by immunohistochemistry and fluorescence in situ hybridization (FISH) using break apart probe for c-MYC gene. For cases with c-MYC rearrangement, additional FISH studies with MYC/IgH fusion probe, BCL2 break apart probe, and BCL6 break apart probe were performed. MYC rearrangement was detected in 23 of 101 cases of aggressive B cell lymphomas (22.5%). The cases with MYC rearrangement were divided into 14 BL, three DLBCL/BL, and six DLBCL. One of the three DLBCL/BL had double translocation for MYC and BCL6 gene. Clinically, DLBCL/BL of the GI tract was more akin to Burkitt lymphoma than it is to GI tract DLBCL with c-MYC rearrangement. The overall prognosis of patients with DLBCL/BL was the worst among aggressive B cell lymphomas. DLBCL/BL is uncommon category of B cell lymphoma in the GI tract with aggressive clinical course despite of intensive chemotherapy.

Kim, Min; Lee, Hee; Lee, Hye; Yang, Han-Kwang; Kim, Woo

doi: 10.1007/s00428-011-1150-6pmid: 21947931

Apoptosis, via caspase cascade, is involved in tumorigenesis and progression, and thus, altered apoptosis-related protein expressions have clinical and prognostic significance. Moreover, the apoptosis pathway is highlighted due to the recent introduction of apoptosis-targeted therapy for several genes such as the X-linked inhibitor of apoptosis protein (XIAP). XIAP is the most potent direct inhibitor of caspase, and XIAP-associated factor 1 (XAF1) and secondary mitochondrial activator of caspase/direct IAP-binding protein with low PI (Smac/DIABLO) are negative regulators of XIAP. In this study, we evaluated the expression of these proteins and investigated their clinical and prognostic significance in gastric carcinomas. Immunohistochemical analysis by using the tissue array method was performed for XIAP, survivin, Bcl-2, XAF1, Smac/DIABLO, and cleaved caspase-3 proteins in 1,162 surgically resected gastric carcinoma cases. XIAP expression was related to the advanced stage. The expression of XIAP showed negative relationship with XAF1 and Smac/DIABLO expressions. In addition, XIAP expression was associated with a poor prognosis and was also proved to be an independent prognostic factor. Cleaved caspase-3 expression was related to the early stage. In addition, cleaved caspase-3 expression was associated with a favorable prognosis and was also proved to be an independent prognostic factor. The expression of XIAP showed an inverse relationship with cleaved caspase-3. In addition, the expression of XAF1 and Smac/DIABLO had a positive relationship with cleaved caspase-3. These findings are consistent with their known functions, and they may help to identify individuals best suited for apoptosis-targeted therapy as a baseline data in gastric carcinoma.

Handra-Luca, Adriana; Olschwang, Sylviane; Fléjou, Jean-François

doi: 10.1007/s00428-011-1152-4pmid: 22002709

The TGFβ signalling pathway is a growth inhibitor system that operates in both normal and tumour cells. Alterations to components of this pathway, including SMAD4, result in resistance to growth inhibition and uncontrolled proliferation. The aim of this study was to analyse the relationships between SMAD4, a key protein in the growth-inhibiting TGFβ pathway; cell proliferation proteins Ki67, p27 and S-phase kinase-associated protein 2 (SKP2); and mismatch repair (MMR) proteins as well as prognostic indicators in colorectal adenocarcinomas. A series of 230 sporadic colorectal adenocarcinomas were studied using tissue microarrays by immunohistochemistry for SMAD4, Ki67, p27, SKP2 and MMR protein (hMLH1, hMSH2 and hMSH6) expression. Protein expression was analysed with respect to pathological prognostic criteria. Loss of SMAD4 nuclear expression (27/230, 12%) correlated with the presence of lymph node metastases, MMR protein expression and the absence of p27 in tumour cells (p = 0.04, p = 0.08 and p = 0.03, respectively). A high Ki67 index did not correlate with SMAD4 expression; however, it did correlate with moderate or poor histological differentiation, SKP2 expression and aberrant or absent MMR protein expression (p = 0.02, p < 0.01 and p < 0.01, respectively). In conclusion, the results of our study suggest that the loss of SMAD4, occurring in 12% of colorectal adenocarcinomas, correlated with the presence of lymph node metastases and absence of p27 expression but not with high cellular proliferation. However, high proliferation correlated with SKP2 and aberrant MMR protein expression. Although the advantage of immunohistochemistry is high throughput, our results allow only an initial evaluation, and subsequent studies, including genetic analyses, are required.

Kreft, Andreas; Kindler, Thomas; Springer, Erik; Kirkpatrick, Charles

doi: 10.1007/s00428-011-1154-2pmid: 22006129

The JAK2-V617F mutation is prevalent in almost all patients with polycythemia vera (PV) and about half of the cases of essential thrombocythaemia (ET) and primary myelofibrosis (PMF). A different allele burden in these entities has long been noticed, but little is known about its distribution among the neoplastic hematopoietic cell lineages within the bone marrow. We conducted a microdissection study of JAK2-V617F-mutated myeloproliferative neoplasms (MPN); 10 cases each of ET, PV, and PMF, with separate analysis of the JAK2 mutation status in three hematopoietic cell lines (i.e., megakaryo-, granulo-, and erythropoiesis). Different numbers of cell lineages harboring the JAK2-V617F mutation were found, being the lowest in ET (17/30), higher in PV (24/30) and in PMF (22/30). The megakaryopoiesis was the most commonly mutated cell lineage (24/30 cases). By analyzing microdissectates we were able to demonstrate a different allele burden of the JAK2-V617F mutation in the megakaryo-, erythro-, and granulopoiesis within the bone marrow of a given case of MPN. We demonstrated differences in the number of mutated cell lineages. The different mutation status may contribute to the different phenotypes of ET, PV, and PMF.

Jenko, Klemen; Kocjan, Boštjan; Zidar, Nina; Poljak, Mario; Strojan, Primož; Žargi, Miha; Blatnik, Olga; Gale, Nina

doi: 10.1007/s00428-011-1139-1pmid: 21912908

Inverted papillomas (IPs) are the most frequent type of sinonasal papillomas. These benign but destructive lesions are known for their high recurrence rate, probably due to incomplete excision. Our aim was to investigate the frequency of human papillomavirus (HPV) infection in patients with IPs and in IPs associated with squamous cell carcinoma (IPsSCC) and to compare it with the frequency of HPV infections in the control group. The influence of HPV infection on the malignant alteration and recurrence rate of IPs was also evaluated. Paraffin-embedded tissue samples from 68 patients with sinonasal IPs and 5 patients with IPsSCC were analyzed in this retrospective study. The control group consisted of 47 patients who had undergone septoplasty or mucotomy of the inferior turbinate. PCR amplification with consensus primer sets was performed to detect alpha-HPVs, and direct sequencing of the PCR products with the same primers was used to determine the HPV genotypes in the samples. We detected HPV DNA in 20 (30.3%) patients with IPs, in 3 (60%) patients with IPsSCC, and in 6 (13%) patients from the control group. The frequency of HPV infection in the study group was statistically significantly higher (p = 0.032) than in the control group. The presence of HPV DNA was not a statistically significant predictor of the recurrence of IPs (p = 0.745) nor was it a statistically significant risk factor for associated SCC (p = 0.32). Since HPV type 11 was the predominant genotype in the IPs, IPsSCC, and in the control cases, we presume that HPV infection may represent incidental colonization rather than being an important etiological factor of IPs.

Matsuyama, Atsuji; Hisaoka, Masanori; Hashimoto, Hiroshi

doi: 10.1007/s00428-011-1149-zpmid: 21927843

Cutaneous mixed tumors, also known as chondroid syringomas, are benign cutaneous adnexal tumors that exhibit remarkable histopathological similarities to pleomorphic adenoma of the salivary gland. Thus far, there is little information on the genetic profiles of cutaneous mixed tumors, although specific genetic aberrations including fusion genes involving PLAG1 and HMGA2 have been demonstrated in pleomorphic adenomas. In the present study, we conducted an immunohistochemical evaluation of PLAG1 and a reverse transcription–polymerase chain reaction (RT–PCR) assay to detect fusion gene transcripts associated with pleomorphic adenoma, including the CTNNB1-PLAG1, LIFR-PLAG1, CHCHD7-PLAG1, TCEA1-PLAG1, HMGA2-FHIT, HMGA2-NFIB, and HMGA2-WIF1 fusion transcripts; this was performed using formalin-fixed paraffin-embedded tumor tissue specimens of 16 cutaneous mixed tumors including one sample with an adenocarcinoma component. All 16 cutaneous mixed tumors were immunoreactive to PLAG1, which was predominantly expressed in cells with myoepithelial or chondroid differentiation accounting for >80% of cells, whereas PLAG1 expression in glandular or squamous tumor cells was restricted to <20% of cells. The carcinoma component in the mixed tumor was also positive for PLAG1. On the other hand, all eight cutaneous adnexal tumors other than the mixed tumor were negative for PLAG1. In RT–PCR analysis, no fusion gene transcripts involving PLAG1 or HMGA2 were identified in any of the cases. Concordant with previous studies, our results support the close relationship between cutaneous mixed tumors and pleomorphic adenomas of the salivary gland. However, the mechanism of PLAG1 expression in cutaneous mixed tumors appears to be possibly different from that of pleomorphic adenomas.

Karlsson, Jenny; Holmquist Mengelbier, Linda; Elfving, Peter; Gisselsson Nord, David

doi: 10.1007/s00428-011-1148-0pmid: 21947875

Wilms’ tumor (WT), the most common kidney tumor among children, is characterized by a triphasic morphology consisting of blastemal, epithelial, and stromal components. Adult WT is a rare malignancy displaying similar histological features. We here present the first published high-resolution genomic analysis of a mixed-type adult WT. This revealed a more pronounced genetic complexity than usually observed in children with mixed-type WT. The majority of chromosomes displayed uniparental disomies, and microdeletions were present in genes with known importance for tumor formation (LRP1B, FHIT, and WWOX) or organogenesis (NEGR1 and ZFPM2), abnormalities not previously reported for pediatric WT. Our results indicate that adult WT is a biological entity distinct from the corresponding pediatric tumor type.