Tumors In Different Groups Research Articles

Increased MIB-1 expression in salivary gland pleomorphic adenoma that recurs and undergoes malignant transformation

The objective of this retrospective study was to explore possible changes in histopathological features and expression of cyclin D1 and MIB-1 in salivary gland pleomorphic adenoma (PA) that recur or undergo malignant transformation. Knowledge of these characteristics might help to guide the management of these rare tumors. The histopathology and immunohistochemical staining characteristics of such tumors were analyzed in a cohort of 65 patients constituting three different groups of tumors: PA, recurrent pleomorphic adenoma (RPA) and carcinoma ex PA (CxPA). The RPAs were divided into two subgroups: primary PA that were known to recur later (PA-prim) and recurrent tumors appearing after a primary tumor (PA-rec). RPAs and CxPAs were compared with PAs without recurrence, which served as a control group. In our study, CxPA and PA-rec, but not PA-prim, showed increased MIB-1 expression compared with the control group. Neither cyclin D1 expression nor any histopathological features showed any association in statistical analyses. CxPA showed increased mitotic activity, squamous metaplasia, and nuclear atypia. Tumor multifocality was more frequent in PA-rec and CxPA. The different MIB-1 expression in CxPA and PA-rec in comparison to PA-prim suggests that the changes in expression could develop after the primary tumor.

Application of One-Step Nucleic Acid Amplification (OSNA) in different cancer entities and usefulness in prostate cancer: a systematic review

BackgroundLymph node (LN) status is a key prognostic factor in the decision-making process of different cancer entities, including prostate cancer (PCa). Sectioning and haematoxylin and eosin (H&E) staining technique remain the gold standard for the evaluation of LN metastases despite some limitations, especially low sensitivity in detecting an accurate tumour burden within the LN, as well as a subjective and time-consuming result. One-step nucleic acid amplification (OSNA) quantifies mRNA copies of cytokeratin 19 (CK19) in a fast, objective, automated, and reproducible way, raising a general interest to explore its utility for lymphatic metastasis identification in different malignancies.MethodsTo present the latest evidence related to the detection of LN metastases in several tumours by using OSNA compared with the conventional H&E method, a systematic review of articles published since March 2021 was conducted using PubMed, Cochrane Library, and Web of Science databases. References from primary papers and review articles were checked to obtain further potential studies. Our procedure for evaluating records identified during the literature search followed the Preferred Reporting Items for Systematic Reviews and Meta-analyses criteria. With the aim to design and justify future clinical routine use of OSNA in PCa, novel PCa evidence has been included in this review for the first time.ResultsTwenty five studies were included. LN from six different groups of tumours: breast, gastrointestinal, gynecological, lung, head and neck and prostate cancers has been assessed. OSNA was compared with post-operative formalin-fixed paraffin-embedded tissue sections with H&E staining as the reference standard. Contingency tables were created, and concordance rate, sensitivity, specificity and predictive values were reported. Seventeen studies analysed the discordant cases using different techniques.ConclusionOSNA method has a high diagnostic accuracy for the detection of LN metastases in several CK19 expressing tumours. Available evidence might encourage future investigations about its usage in PCa patients to improve LN staging and prognosis.

Immunotherapies in ovarian cancer

Ovarian cancer is the leading cause of death for gynaecological cancer, and new therapies are urgently awaited. Although the presence of tumour-infiltrating lymphocytes has been confirmed to be associated to a better prognosis, immunotherapy is not yet incorporated to the armamentarium in ovarian cancer. This review briefly summarises the strategies that have been tested or are under study for the three different groups of tumours: immune desert, inflamed and immune-excluded ovarian tumours. Finally, a better knowledge of the biology and immune microenvironment is needed for successfully developing new immunotherapy strategies.

Impact of the Updated 2018 American Society of Clinical Oncology/College of American Pathologists Guideline for Human Epidermal Growth Factor Receptor 2 Testing in Breast Cancer.

The updated American Society of Clinical Oncology/College of American Pathologists guideline for human epidermal growth factor receptor 2 (HER2) testing in breast cancer requires pathologists to re-evaluate HER2 status. To define HER2 status of breast cancer using immunohistochemistry and fluorescence in situ hybridization. Diagnostic reports of invasive breast cancers made between 2014 and 2018 with HER2 immunohistochemistry and fluorescence in situ hybridization results were retrieved. HER2 status was re-defined using the updated recommendations. Of 2514 tumors, 89.7% (2254 of 2514) suggested for fluorescence in situ hybridization assay were HER2 immunohistochemistry 2+. Approximately 8.9% (225 of 2514) and 1.4% (35 of 2514) of tumors were of immunohistochemistry 0/1+ and 3+, respectively. Based on the average HER2 copy number and HER2:CEP17 ratio, tumors were assigned into 5 groups, including 13.1% (330 of 2514) group 1 tumors, 2.1% (52 of 2514) group 2 tumors, 1.1% (27 of 2514) group 3 tumors, 7.0% (175 of 2514) group 4 tumors, and 76.8% (1930 of 2514) group 5 tumors. In combination with immunohistochemistry, all tumors in group 2 and group 4 changed HER2 status, from positive and equivocal into negative, respectively, while group 3 tumors remained positive. Comparative analyses of clinicopathologic features of tumors in different groups revealed that group 2 and 4 tumors displayed worse clinicopathologic features than those of group 5, while group 3 tumors shared similar clinicopathologic features to those of group 1. Following the updated guideline, HER2 status is clearly designated. Significant differences regarding clinical features were observed between tumors in different groups but they share the same HER2 status, suggesting further validation of the accuracy of this diagnostic approach is warranted.

Structural characterization of a low molecular weight polysaccharide from Grifola frondosa and its antitumor activity in H22 tumor-bearing mice

• A low molecular weight polysaccharide (LMw-GFP) was isolated from Grifola frondosa by ultrafiltration. • Structural characteristics of LMw-GFP were identified by HPGPC, MS, FT-IR, GC, NMR and methylation analysis. • LMw-GFP could inhibit the growth of solid tumor via activating anti-tumor immune responses in H22 tumor-bearing mice. • The apoptotic properties of H22 solid tumors in different groups were further investigated. A novel low molecular weight polysaccharide from Grifola frondosa (LMw-GFP) was prepared in this study, and its structural characteristics and antitumor mechanism in vivo were further investigated. The results showed that LMw-GFP contained 95.83% carbohydrate, 0.34% protein and 0.21% uronic acid, with the average molecular weight of approximately 1.79 × 10 3 Da. Additionally, FTIR, GC, NMR and methylation analysis further demonstrated that LMw-GFP was a homogeneous polysaccharide composed of α-T-Glc p (28.26%), α-1 → 4-Glc p (50.24%) and α-1 → 3,4-Glc p (21.50%). The in vivo antitumor experiments showed that LMw-GFP exhibited antitumor effects on H22 hepatoma cells with an inhibitory ratio of 40.1% via protecting immune organs and enhancing the activities of NK cells, macrophages and lymphocytes. Besides, the H&E staining, cell cycle analysis and fluorescent staining results showed that LMw-GFP could induce H22 solid tumor cells apoptosis in vivo via mitochondrial apoptotic pathway.

PO-460 Gene expression-based probabilistic graphical models identify three independent biological layers in colorrectal cancer

IntroductionColorectal cancer (CRC) is the fourth cause of cancer death in the world. The Colorectal Cancer Subtyping Consortium (CRCSC) defined four consensus molecular subtypes (CMS): immune, canonical, metabolic and mesenchymal. This study bases on the hypothesis that there are two independent biological layers, as our group has previously seen in other malignancies, such as Triple negative breast cancer, melanoma or bladder cancer. So, the aim of this studio is to put the colorrectal cancer subtypes in context with that hypothesis.Material and methodsColorectal cancer tumour datasets were obtained from Gene Expression Omnibus (GEO). A merged database was constructed, and limma R package was used to remove batch effect. A probabilistic graphical model, followed by successive sparse k-means and consensus cluster analyses were used to classify CRC tumour samples from the merged dataset.Results and discussionsThrough this approach, three independent classifications, immune, molecular and adhesion-related, were established. Molecular classification showed different groups of tumours that present differential expression of diverse colorectal biomarkers. The immune classification further divided patients into high immune activity and low immune activity subgroups. And the adhesion-related assignment divides patients in two subgroups with prognostic value. We showed that immune and adhesion-related layers, when studied together, add its prognostic value defining low risk and high risk subgroups. We finally compare our classifications with the consensus molecular classification.ConclusionMolecular information obtained using a novel analytical approach to classify colorectal cancer tumour samples allowed us to establish new classifications based on different biological layers. We have demonstrated that our classifications based on the immune and the adhesion-related layers have prognostic value. Ultimately, to deepen the knowledge of CRC makes it possible to propose new therapeutic strategies and could also be used for diagnostic and prognostic purposes.

Abstract ES11-3: Novel approaches for selection of and monitoring treatment

Abstract With the introduction of new therapeutic options in advanced and metastatic breast cancer, there is a growing demand for new biomarkers to guide treatment decisions. There are several differences compared to early breast cancer that have to be addressed to define the clinical use of precision medicine in advanced and metastatic breast cancer. The current challenges include the limited availability of tissue in metastatic disease, the problems of spatial heterogeneity in large advanced tumors, as well as differences between different metastatic sites. In addition, it has been shown that in metastatic and recurrent tumors, new molecular alterations develop under therapeutic pressure, for example mutations in the ESR1 receptor. The current guidelines suggest that the basic characterization of the tumor using the standard markers of hormone receptors and HER2 should be repeated in the metastatic setting, if possible. Differences between primary and recurrent tumors have been observed in a comparably small percentage of tumors, nevertheless these differences may open new therapeutic options. Therefore, rebiopsies should be performed, if possible, these biopsies are also highly relevant for the evaluation of new therapy strategies in clinical trials. Another relevant challenge is the heterogeneity of the tumor cells at different metastatic sites, these different metastases originate from different cell clones and have therefore a different molecular background, in particular in those tumors that have a high intratumoral molecular diversity. Considering this heterogeneity between different metastases as well as technical difficulties in the conduction of biopsies, new technologies have been established to monitor tumor biomarkers on a systemic level. This includes the evaluation of circulating tumor cells as well as the evaluation of cell-free circulating DNA (cfDNA) in so called liquid-biopsies. Several translational studies shown that sequencing of cfDNA is suitable to detect tumor mutations that can be used distinguish different groups of tumors with different response rates, for example to PIK3CA inhibitors. Relevant mutations include ESR1 and PIK3CA, which are relevant for therapy resistance in breast cancer and can be measured by NGS-sequencing. In breast cancer, these investigations are currently not part of the clinical routine. In lung cancer, the analysis of T790M EGFR mutations in liquid biopies are routinely used to guide therapeutic decisions in recurrent disease, which shows that the introduction into the clinical setting is feasible. The presentation will discuss the advantages and limitations of the different methods, and give examples for biomarker strategies for different therapies including endocrine therapy, immune therapy and targeted therapy. Citation Format: Denkert C. Novel approaches for selection of and monitoring treatment [abstract]. In: Proceedings of the 2017 San Antonio Breast Cancer Symposium; 2017 Dec 5-9; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2018;78(4 Suppl):Abstract nr ES11-3.

Similar length of colon is removed regardless of localization in right‐sided colonic cancer surgery

Colorectal cancers represent a heterogenous group of tumours. While left segmental colectomy is an accepted and oncologically safe practice for left-sided colonic cancer (CC), some authors suggest that limited segmental resection of right-sided cancer should be debated in order to preserve length of the resected colon. To our knowledge, caecum and ascending CC have not been analysed as different groups of tumours. The objective of this study was to assess if, retrospectively, surgical treatment of caecal cancer differed from ascending CC. A review of all consecutive patients with right colonic resection for cancer admitted to the University hospital of Grenoble from January 2005 to August 2016 was performed. Length of resected colon was compared between caecal primary and ascending CC. Other technical and pathological aspects were analysed such as minimal invasive surgery and number of harvested lymph nodes from anatomic specimens. Among operated patients, tumour was localized pre-operatively on caecum in 110 cases and on the right ascending colon in 119 cases. Pre-operative localization had no effect on resected colon length (mean 24.5 cm, 24 ± 10.34 versus 25 cm ± 7.28, P = 0.95), on the number of harvested lymph nodes with a mean of 15 (±7.6) nodes in the caecal group versus 15.2 (±7.3) (P = 0.72). We noticed 15 cases of discordance between pre- and post-operative localization (4 versus 11, P = 0.08). Length of resected colon does not differ depending on localization of tumour in our center.

Mutant-Allele Tumor Heterogeneity Scores Correlate With Risk of Metastases in Colon Cancer

BackgroundColorectal cancer is a leading cause of cancer-related mortality, has a very broad mutational spectrum, and there is no clinically available biomarker that can predict which patients with stage II or stage III colorectal cancer will develop metastatic disease. Patients and MethodsWe used a targeted next-generation sequencing approach to analyze the mutational spectra in stage II and III colon cancer patient samples. ResultsAmidst a broad range of acquired mutations and variants, we found evidence of tumor heterogeneity that distinguished the tumors in different groups. When heterogeneity was quantified using the Mutant-Allele Tumor Heterogeneity (MATH) score, there was a strong correlation between higher MATH score and risk of metastases. ConclusionsMeasures of tumor heterogeneity might be useful biomarkers for identifying patients with colon cancer who are at risk of developing metastases. This might allow for more specific, tailored follow-up and adjuvant therapies after standard surgery.

The use of matrigel has no influence on tumor development or PET imaging in FaDu human head and neck cancer xenografts.

BackgroundIn preclinical research MatrixgelTM Basement Membrane Matrix (MG) is used frequently for the establishment of syngeneic and xenograft cancer models. Limited information on its influence on parameters including; tumor growth, vascularization, hypoxia and imaging characteristics is currently available. This study evaluates the potential effect of matrigel use in a human head and neck cancer xenograft model (FaDu; hypopharyngeal carcinoma) in NMRI nude mice. The FaDu cell line was chosen based on its frequent use in studies of cancer imaging and tumor microenvironment.MethodsNMRI nude mice (n = 34) were divided into two groups and subcutaneously injected with FaDu cells in medium either including (+MG) or excluding matrigel (−MG). In sub study I seven mice from each group (+MG, n = 7; −MG, n = 7) were 18F- fluorodeoxyglucose (18F-FDG) PET/CT scanned on Day 5, 8, 12, 15, and 19. In sub study II ten mice from each group (+MG, n = 10; −MG, n = 10) were included and tumors collected for immunohistochemistry (IHC) analysis of tumor microenvironment including; proliferation ratio, micro vessel density, average vessel area, hypoxia, nuclear density, and necrosis. Tumors for IHC were collected according to size (200–400 mm3, 500–700 mm3, 800–1100 mm3).ResultsFDG uptake and tumor growth was statistically compatible for the tumors established with or without MG. The IHC analysis on all parameters only identified a significantly higher micro vessel density for tumor size 500–700 mm3 and 800–1100 mm3 and average vessel area for tumor size 500–700 mm3 in the −MG group. Comparable variations were observed for tumors of both the +MG and −MG groups. No difference in tumor take rate was observed between groups in study.ConclusionsMatrigel did not affect tumor growth or tumor take for the FaDu xenograft model evaluated. Tumors in the -MG group displayed increased angiogenesis compared to the +MG tumors. No difference in 18F-FDG PET uptake for tumors of different groups was found. Based on these observations the influence of matrigel on tumor imaging and tumor microenvironment seems minor for this particular xenograft model.

Diffusely infiltrating astrocytomas: pathology, molecular mechanisms and markers.

Diffusely infiltrating astrocytomas include diffuse astrocytomas WHO grade II and anaplastic astrocytomas WHO grade III and are classified under astrocytic tumours according to the current WHO Classification. Although the patients generally have longer survival as compared to those with glioblastoma, the timing of inevitable malignant progression ultimately determines the prognosis. Recent advances in molecular genetics have uncovered that histopathologically diagnosed astrocytomas may consist of two genetically different groups of tumours. The majority of diffusely infiltrating astrocytomas regardless of WHO grade have concurrent mutations of IDH1 or IDH2, TP53 and ATRX. Among these astrocytomas, no other genetic markers that may distinguish grade II and grade III tumours have been identified. Those astrocytomas without IDH mutation tend to have a distinct genotype and a poor prognosis comparable to that of glioblastomas. On the other hand, diffuse astrocytomas that arise in children do not harbour IDH/TP53 mutations, but instead display mutations of BRAF or structural alterations involving MYB/MYBL1 or FGFR1. A molecular classification may thus help delineate diffusely infiltrating astrocytomas into distinct pathogenic and prognostic groups, which could aid in determining individualised therapeutic strategies.

Significance of immunohistochemistry method for diagnosing of stomach tumors

The article is dedicated to the diagnosis of malignant tumors of the stomach, analyzing divergences of diagnoses after routine morphological researches and immunomorphological methods with monoclonal antibodies using, assessment of proliferative activity of different groups of tumors of this localization. Our investigation demonstrated that in the group of gastric tumors mistaken primary diagnoses were identified in 33 of 62 cases (53.2%). Proliferative score estimation is too important to determine the potential for malignancy of gastrointestinal stromal tumors and the degree of differentiation of neuroendocrine cancers.

Gemcitabine plus cisplatin neoadjuvant chemotherapy with radical cystectomy for the treatment of muscle invasive bladder cancer: 9 years retrospective analysis

Objective To assess the value of gemcitabine plus cisplatin (GC) neoadjuvant chemotherapy with radical cystectomy in the treatment of patients with muscle invasive bladder cancer (MIBC).Methods The data of 114 patients with MIBC received GC neoadjuvant chemotherapy with radical cystectomy from December 2003 to December 2012 were retrospectively compared with those in other 120 patients treated with radical cystectomy alone from June 2003 to December 2012 in our institution.There was no significant difference of the gender,age and stage between the 2 groups (P＞0.05).The duration of free survival,overall survival and the local control of tumor in different groups were compared.Results In the chemotherapy group,76 cases were found downstaging of the tumor after the operation,including 27 cases with tumor free.The mean duration of following up was (51 ±25) months.Forty-three patients died during the follow-up period.The median survival was 79 months and the 2-year recurrence free rate was 75.9％.In surgery alone group,23 cases were found downstaging of the tumor,including 5 cases with tumor free.The patients were followed up for (38±20) months.Fifty-seven patients died during the following up.The median survival was 44 months and the 2-year recurrence free rate was 62.8％.There were significant differences in downstaging rate of the tumor,recurrence free survival and overall survival between the 2 groups (P＜0.05).Conclusion GC neoadjuvant chemotherapy can improve survival of the patients with MIBC,which facilitate to downstage the tumor and increase the recurrence free survival duration and overall survival duration. Key words: Urinary bladder neoplasms; Muscle invasive; Antineoplastic combined chemotherapy protocols; Neoadjuvant chemotherapy; Gemcitabine plus cisplatin

El papel de la metaloproteinasa de la matriz MMP-9 y del inhibidor tisular de metaloproteinasa TIMP-2 como marcadores séricos de cáncer vesical

IntroductionThe diagnosis and molecular staging of bladder cancer based on the detection of gelatinases mRNA (MMP-2 and MMP-9) in peripheral blood circulating and mononuclear cells have shown promising results. We analyze if the determination of the corresponding protein synthesis products makes it possible to diagnose and characterize patients with bladder cancer. Material and methodQuantification of the serum levels of MMP-2, MMP-9 and TIMP-2 in a series of 42 individuals (31 patients with bladder cancer in different stages and 11 healthy controls) using the ELISA technique was carried out. The determinations were compared between cases and controls (Mann-Whitney U) and between different groups of tumors (Mann-Whitney U or Kruskal-Wallis), according to the clinical-pathological characteristics (age, gender, T category, M category or grade). Diagnostic yield of these markers was evaluated by analysis of the ROC curves. ResultsThere is a correlation between the determinations of MMP-2 and TIMP-2 (R=.699; P>.0001) and MMP-9 and TIMP-2 (R=.305; P=.049). Patients with bladder cancer have higher levels of MMP-9 (p<0.0001) and TIMP-2 (P=.047) than the controls. Furthermore, the MMP-9/TIMP-2 ratio is also superior in cancer patients (P<.001). Differences were not detected between cancer and controls regarding age (P=.64) or gender (P=.64). Differences were also not detected regarding MMP-2 (P=.35) or MMP-2/TIMP-2 rate (P=.45). Within the cancer patient population, the MMP-2 and MMP-9 values differ according to T category (P=.022 and P=.038, respectively) and those of the TIMP-2 according to M category (P=.036). ROC curve analysis showed that both MMP-9 and the MMP-9/TIMP-2 ratio discriminate patients with cancer and controls, with equivalent diagnostic accuracy (ABC 0.953) and cut offs of 3.93 ng/mL (S 90%; Sp 81%) and 0.053 ng/mL (S 96%; Sp 84%), respectively. ConclusionsThe results obtained suggest that both serum MMP-9 and TIMP-2 would have an application in the prediction of the development and progression of bladder cancer, and a potential utility as clinical markers of the disease. Multicenter, prospective studies that confirm their preliminary results are necessary.

Meta-analysis of Green Tea Drinking and the Prevalence of Gynecological Tumors in Women

We performed a meta-analysis of the correlation between drinking green tea and the risk of female ovarian tumors. Related literature (2000-2010) was retrieved from PubMed, EMbase, CBMdisc, CNKI, and Wanfang databases. The relationship between the prevalence of ovarian cancer and drinking tea in cohort studies was explored. RevMan5.1.0 software was used for the meta-analysis. A total of 6 case control studies and cohort studies were included. A total of 9113 participants, 3842 cases, and 5271 control cases were included in our analysis. Our analysis indicates that drinking green tea can significantly decrease the risk of ovarian cancer (odds ratio = 0.81; 95% confidence interval = 0.73-0.89; P < .0001). Further research is needed to explore the relationship between drinking green tea and the risk of ovarian tumor in different groups of people and with different tea types and dosages.

Expression and methylation profiles associated with recurrent mutations in stage II colorectal cancer.

e14613 Background: Aberration of normal genetic and epigenetic patterns occurs at early stages of colorectal cancer (CRC) and accumulates throughout cancer progression. To characterize pre-metastatic tumors, a series of stage II, microsatellite stable, colon tumors and their paired mucosa were profiled on RNA expression and DNA methylation microarrays ( www.colonomics.org ). Our aim is to define molecular subtypes based on recurrent gene mutations, methylation and expression profiles, and explore if these molecular subtypes are associated to patients’ prognosis. Methods: We have sequenced exomes (Illumina Genome Analyzer) of a subset of 42 COLONOMICS normal-tumor paired samples (21 good and 21 bad prognosis). Variants identified in normal tissue were used to filter SNPs. DNA methylation (Infinium Human Methylation 450k) and RNA expression (Affymetrix U219) data from those samples were used in this analysis. Correlation was used to assess the association between tumor mutations and differentially expressed/methylated genes. Significant genes were subsequently used to perform tumor clustering. Results: Exome analysis revealed a mean of 150 somatic mutations per sample. From these, 12 variants were recurrently mutated (KRAS, TP53, etc) in more than 3 tumors that were used to define tumor subtypes based on gene methylation/expression patterns. We obtained 12 profiles that clearly identified the cluster of mutated samples. For some profiles, the cluster only includes those tumors with the mutation. Interestingly, some clusters included the mutated samples and additional tumors showing the same phenotype despite not having the mutation. For each mutation, the overlapping between the differently methylated/expressed genes ranged from 14 to 200 common genes. When combining data from the three platforms two main CRC molecular subtypes emerge; each of which shows molecular heterogeneity but no association with prognosis. Conclusions: Mutational status is associated with gene methylation and expression patterns in CRC patients. Although none of these clusters was associated with prognosis, different groups of tumors could be related to distinctive pathways, which may reveal useful as therapeutic targets.

Use of brightness wavelet transformation for automated analysis of serum metallothioneins‐ and zinc‐containing proteins by Western blots to subclassify childhood solid tumours

In this study, we determined serum levels of metallothioneins (MTs) and zinc in children with solid tumours (neuroblastoma, Hodgkin lymphoma, medulloblastoma, osteosarcoma, Ewing sarcoma and nephroblastoma) by differential pulse voltammetry Brdicka reaction and ELISA. Zn(II) level in patients sera was 40% compared to controls, contrariwise, MT level was 4.2 × higher in patients. No significant differences among single diagnoses were found both for Zn(II) and MT. When determined Zn(II)/MT ratio, in controls its value was 24.6, but it was 2.6 in patients. After Western-blotting with anti-MT and anti-Zn chicken antibodies, variable intensities of the bands within the samples were observed. The brightness curve obtained for each sample both for MT- and Zn blots was further analysed to produce a list of band positions together with some complementary information related to the intensity of the observed bands by the optimised algorithm. We constructed from those profiles decision trees that enable to distinguish different groups of tumours. The blood samples were heat-treated, in which we supposed mainly MT, but samples contained other thermostable Zn-containing proteins that were helpful for identification of embryonal tumours with 88% accuracy and for identification of sarcomas with 78% accuracy. In MT blots the accuracies were 53 and 45%, respectively. Simultaneous analysis of MT and Zn blots did not increased accuracy of identification neither in embryonal tumours (80%) nor in sarcomas. Those results are promising not only from diagnostic point of view but particularly in the area of studying of individual MT isoforms and their aggregates in malignant tumours.

Comparison of the Planning Target Volume Based on 3-dimensional CT and 4-dimensional CT Images of Thoracic Esophageal Cancer

To compare the centroid position, volumetric differences and dice similarity coefficient (DSC) of planning target volume (PTV) based on three-dimensional CT (3DCT) and four-dimensional CT (4DCT) for the thoracic primary tumor of esophageal cancer. Forty-three patients with esophageal cancer underwent 3DCT and 4DCT simulation scans of the thorax during normal free breathing. The motion of primary tumors located in the proximal (group A), mid-(group B), and distal (group C) thoracic esophagus were obtained from the 4DCT scans. PTV3D was defined on 3DCT using the tumor motion measured by 4DCT scans; PTV4D was defined as the union of the target volume contoured on the 10 phases of 4DCT images. The differences in target centroid position, volume, DSC, and degree of inclusion (DI) between PTV3D and PTV4D were evaluated for tumors in different groups respectively. The mean centroid shifts between PTV3D and PTV4D in the three dimensional directions were less than 0.3 cm, for all the three groups. However, the target centroid shift was demonstrated significant difference in lateral direction for group C (Z = -2.355, p = 0.019). The median size ratio of PTV4D to PTV3D was 0.79 ± 0.07, 0.91 ± 0.25, and 0.70 ± 0.07 for group A, B and C. There was no significant correlation between the ratio of PTV4D to PTV3D and the target motion for the three groups. The DSC between the PTV4D and PTV3D were 0.87 ± 0.04, 0.88 ± 0.06, and 0.81 ± 0.03. The median DI of PTV3D in PTV4D was 0.78, 0.85, 0.69 for group A, B and C, and the median DI of PTV4D in PTV3D was 0.98, 0.97, and 0.99 for group A, B and C, respectively. The 3DCT-based PTV provides a good coverage of the 4DCT-based PTV, especially for middle esophageal tumors, but for proximal and distal esophageal tumors, PTV3D also encompasses relatively large normal tissues.

Immunohystochemical analysis of SOCS3 in prostatic adenocarcinoma biopsies.

e15132 Background: Chronic inflammation and the subsequent tissutal alterations may play a key role in prostate carcinogenesis. Molecular alterations of the suppressor of cytokine signaling (SOCS)3, can contribute to explain the pleiotropic role of interleukin (IL)-6 in this type of cancer. In a recent study, we demonstrated that methylation of SOCS3 may be involved in the pathogenesis of prostate cancer and could identify in a subset of tumor with an aggressive behavior. Methods: We analyzed the protein expression of SOCS3 by immunohistochemistry in 50 prostate cancer biopsies: 30 prostatic carcinoma with Gleason score 7, 10 with Gleason score <7 and 10 with Gleason score >7. Slides were incubated with monoclonal antibody SOCS3 (1E4) (1.5 mg/ml; Abnova, Taiwan). The SOCS3 staining intensity were evaluated by two pathologist (FP and LML) in three different way: positive; negative; weak intensity staining with negative neoplastic glands. Colonic mucosa were used as positive control. Results: SOCS3 positive were found in all cases of adenocarcinoma Gleason score < 7 and in 10 cases with Gleason score 7, negative staining were found in all cases of adenocarcinoma Gleason score > 7 and in 8 cases with Gleason score 7. Weak SOCS3 staining with presence of negative neoplastic glands were found in 12 cases of adenocarcinoma Gleason score 7. The group of tumor SOCS3 negative were significantly more aggressive than group SOCS3 positive in terms of TNM stage,positivity of surgical margin. Cases with weak SOCS3 staining and negative neoplastic glands displayed more aggressive clinical behavior than cases SOCS3 positive (increase number of T3a and positive surgical margins), on the other hand unlike SOCS3 negative no stage T3b and lymphnode metastasis were found. Conclusions: Immunohystochemical analysis of SOCS3 protein unlike methylathion study identify three different groups of tumors: positive, negative and with weak SOCS3 staining with the presence of negative neoplastic glands. In this last group the reduction of expression protein and the presence of negative neoplastic glands could indicate an intermediate stage of tumor turning towards shutdown of SOCS3 and thus object of strict follow-up and suitable to demethylating agents therapy.

CD10 (Neutral Endopeptidase) Expression in Myoepithelial Cells of Salivary Neoplasms

CD10 is a cell surface peptidase expressed in a wide variety of normal and neoplastic tissues, including breast myoepithelial cells. In salivary glands, expression of CD10 has only been used to identify neoplastic myoepithelial cells of pleomorphic adenomas and myoepithelial carcinomas. However, its accuracy in other salivary tumors with myoepithelial component has yet to be analyzed. We examined 72 salivary tumors with myoepithelial differentiation using immunohistochemical technique to detect CD10. In salivary glands, CD10 expression was not detected in myoepithelial cells. Only fibrocytes within the intralobular stroma were CD10 positive. In neoplastic myoepithelial cells, CD10 expression was found in 25.71% of benign and 32.43% of malignant neoplasms. When the different groups of tumors were compared, epithelial-myoepithelial carcinomas (EMEC) showed a stark contrast with the others (83.3% of cases with CD10 expression). Surprisingly, adenoid cystic carcinomas and basal cell adenomas were negative in 100% of the cases. Myoepitheliomas, pleomorphic adenomas, and myoepithelial carcinomas were positive in 27.7%, 30.0%, and 40% of the cases, respectively. In conclusion, salivary neoplastic myoepithelial cells gain CD10 expression in relation to their normal counterparts. However, the gain of this protein is not a sensitive marker for detecting myoepithelial cells in the majority of the tumors, except for EMEC. The high expression of CD10 by this carcinoma can be a valuable tool to separate EMEC from the tubular variant of adenoid cystic carcinomas in small incisional biopsies, where the precise diagnosis may be impossible.