Degree Of Cellular Differentiation Research Articles

Postmenopausal Ovarian Sertoli-Leydig Tumours: Case Report

Sertoli and Leydig cell tumours are rare secretory tumours of the mesenchyme and sex cords. Diagnostic certainty is histological after surgery and there is no specific ultrasound sign despite the strong clinical presumption. The prognosis depends on the degree of cellular differentiation and the presence of heterologous elements. We report on the occurrence of a sertoli-leydig tumour in a 50-year-old postmenopausal woman. Poorly differentiated forms of Sertoli-Leydig tumours have a significant malignant potential. Treatment is surgical, with chemotherapy providing an interesting adjuvant. The prognosis after surgery is dominated by recurrence.

The Diagnostic Role of FGF 21 in Endometrial Cancer and Other Pathologies of the Uterine Corpus.

Endometrial cancer is becoming an increasing problem. Taking into account its pathomechanisms, we aimed to investigate whether FGF 21, an important metabolism regulator, could be used as a biomarker for endometrial cancer. The study included 233 patients who were classified into five subgroups depending on the result of the histological examination: endometrial carcinomas, sarcomas, endometrial polyps, fibroids, and normal endometrium. Statistically significantly higher FGF 21 levels were found in patients diagnosed with malignant lesions (p < 0.001). FGF 21 concentration correlated with the degree of cellular differentiation (p = 0.020) and the presence of lymph node metastases (p = 0.009). The diagnostic performance characteristics of FGF 21 as an EC diagnostic marker demonstrated an AUC of 0.677. Of all of the assessed biomarkers, FGF 21 had the highest specificity (90%), yet limited sensitivity (41%). Additionally, HE4 and CA 125 were confirmed to have roles as EC biomarkers, with a higher accuracy for HE4 (79% vs. 72%).

Interleukin-34 deficiency aggravates development of colitis and colitis-associated cancer in mice

Although expression of interleukin (IL)-34 is upregulated in active ulcerative colitis (UC), the molecular function and underlying mechanism are largely unclear. To investigate the function of IL-34 in acute colitis, in a wound healing model and in colitis-associated cancer in IL-34-deficient mice. Colitis was induced by administration of dextran sodium sulfate (DSS), and carcinogenesis was induced by azoxymethane (AOM). Whether the impact of IL-34 on colitis was dependent on macrophages was validated by depletion of macrophages in a murine model. The association between IL-34 expression and epithelial proliferation was studied in patients with active UC. IL-34 deficiency aggravated murine colitis in acute colitis and in wound healing phase. The effect of IL-34 on experimental colitis was not dependent on macrophage differentiation and polarization. IL-34-deficient mice developed more tumors than wild-type mice following administration of AOM and DSS. No significant difference was shown in degree of cellular differentiation in tumors between wild-type and IL-34-deficient mice. IL-34 was dramatically increased in the active UC patients as previously reported. More importantly, expression of IL-34 was positively correlated with epithelial cell proliferation in patients with UC. IL-34 deficiency exacerbates colonic inflammation and accelerates colitis-associated carcinogenesis in mice. It might be served as a potential therapeutic target in UC.

Notch Signaling in Thyroid Cancer.

Thyroid cancer is the most common malignancy of the endocrine system with a steadily rising incidence. The term "thyroid cancer" encompasses a spectrum of subtypes, namely papillary thyroid cancer, follicular thyroid cancer, anaplastic thyroid cancer, and medullary thyroid cancer. Each subtype differs histopathologically and in degrees of cellular differentiation, which may be in part due to signaling of the Notch pathway. The Notch pathway is an evolutionarily conserved signal transduction mechanism that regulates cell proliferation, differentiation, survival, stem cell maintenance, embryonic and adult development, epithelial-mesenchymal transition, and angiogenesis. Its role in cancer biology is controversial, as it has been shown to play both an oncogenic and tumor-suppressive role in many different types of cancers. This discordance holds true for each subtype of thyroid cancer, indicating that Notch signaling is likely cell type and context dependent. Whether oncogenic or not, Notch signaling has proven to be significantly involved in the tumorigenesis of thyroid cancer and has thus earned interest as a therapeutic target. Advancement in the understanding of Notch signaling in thyroid cancer holds great promise for the development of novel treatment strategies to benefit patients.

Effects of COJEC induction on neuroblastoma patient-derived xenografts (PDX).

e21503 Background: Neuroblastoma (NB) is a solid pediatric tumor that causes 15% of childhood cancer deaths. High-risk patients have 50 to 70% mortality after 5 years. Currently, the standard way to treat high risk patients is COJEC induction followed by surgery, chemotherapy with stem cell transplants, irradiation and immunotherapy. This intense treatment has significantly increased overall survival, but mortality is still high, survivors are prone to relapse and treatment resistance is a major clinical problem. COJEC induction consists in the cycled administration of the chemotherapeutics Cisplatin, Vincristine, Etoposide, Cyclophosphamide and Carboplatin. These drugs have been long used in the clinic, but a comprehensive preclinical study combining all of them had not been done. The aim of this project was to establish a PDX model for chemotherapy induction in high-risk NB and investigate tumor characteristics and response upon treatment. Methods: NB cells from previously developed high-risk PDX models were injected subcutaneously in nude mice. When tumors reached an appropriate size, they were treated with either a COJEC-like protocol or single agent induction with high dose cisplatin. After treatment, tumors were analyzed by immunohistochemistry, chromosomal copy number analysis and RNA sequencing. Results: Most mice responded well to COJEC induction and 8/10 tumors were reduced. Within the reduced tumors, 4 were subjected to total resection and relapsed after several weeks, mimicking the clinical situation. Cisplatin treatment alone led to no significant tumor reduction and 6/10 tumors were highly resistant. Thus, as for humans, multidrug induction is more successful in reducing tumor size but presents an uneven response with high relapse rate. The responsive tumors show a higher degree of cellular differentiation when compared to resistant tumors. Cisplatin treated tumors have accumulation of chromosomal aberrations when compared to controls, and subclonal dynamics are enhanced in resistant tumors. Conclusions: We established a PDX model for chemotherapy induction in high-risk NB. The results mimic clinical findings seen in high-risk NB patients. The model is useful to understand the mechanisms of treatment resistance and for the testing of novel therapies against high-risk NB.

PS02.021: TOTALLY LAPAROSCOPIC TRANSHIATAL ESOPHAGECTOMY FOR ADENOCARCINOMA OF THE ESOPHAGOGASTRIC JUNCTION: INDICATIONS, TECH. ASPECTS AND RESULTS, COMPARING TO OPEN ACCESS

Abstract Background Minimally invasive esophagectomy remains with high morbidity and mortality rates. The transhiatal technique has less surgical time, lower morbidity and an acceptable oncological surgical control for patients with adenocarcinoma of the esophagogastric junction (AGEJ), unfortunaly still infrequent by laparoscopy. The aim of this study is to present indications, technical aspects and results of the totally laparoscopic transhiatal esophagectomy (TLTE) comparing to open access (OA) Methods This study retrospectively reviewed esophageal cancer patients that were admitted at an oncology referral center between 2011 to 2016. The studied population was composed of 128 AEGJ, all of them characterized as Siewert I and II classification. Demographic, pathological and clinical characteristics were analyzed and compared to clinical stage and overall survival at 60 months, between TLTE and OA. Influence of prognostic variables was assessed by Cox regression. Results The mean age was 63,4 ys (55–80ys), 85,4% were male. The selection of this group of patients was related to good clinical condition, however most of T2-T3 tumors. 90 patients were submitted to OA and 38 to TLTE (p-value for Log-Rank 0.11). Average follow-up was 39.8 months. By univariate analysis, curative intention resection was related to only 22% of patients in 5 ys. By univariate Cox regression, only degree of cellular differentiation was related to poor clinical stage in TLTE (p-value = 0.05). Weight loss (kg), BMI variation (kg/m²) and percentage of weight loss from initial symptoms to the diagnosis of esophageal carcinoma are factors that predict indicate the TLTE. Considering only patients submitted to curative intent surgery, more than 21 node resection could reach a statistically significant improvement in survival rate by univariate analysis, but there were no difference between both groups.There was no mortality and morbidity rate about 7%. Comparing TLTE to OA there were no difference in survival and morbidity and mortality. Conclusion This is a preliminary study about low rate related to both access (TLTE and OA) in patients with AEGJ) with T2-T3 tumors, with low morbidity and no mortality series. TLTE is a safe and feasible alternative to the open approach, with decrease short-term morbidity with comparable oncological outcomes. Disclosure All authors have declared no conflicts of interest.

Estudio longitudinal: Cáncer de Ovario en el Instituto Nacional de Oncología y Radiobiología de Cuba (INOR), resultados del tratamiento.

Introducción: El cáncer de ovario es una causa común de muerte entre las mujeres que desarrollan neoplasias ginecológicas. La supervivencia depende de factores clínicos y del tipo de cirugía. El objetivo del presente estudio es describir la supervivencia de un grupo de pacientes con diagnóstico de cáncer de ovario tomando en cuenta varios factores.&#x0D; Métodos: El presente estudio longitudinal retrospectivo se realizó en mujeres con cáncer de ovario tratadas en el Instituto Nacional de Oncología y Radiobiología, La Habana-Cuba entre el 1 de enero y el 31 de diciembre de 2005, con seguimiento de por lo menos 5 años. Las variables fueron edad, tipo histológico del tumor, etapa clínica de la FIGO, grado de diferenciación celular, valores del biomarcador CA-125, tipo de cirugía realizada, respuesta a la quimioterapia utilizada, supervivencia, recurrencias. El paquete estadístico utilizado fue SPSS 11.0 para Windows. Se aplicaron pruebas estadísticas como la prueba no paramétrica de Kruskal-Wallis, la prueba no paramétrica de MannWhitney y la prueba de Chi-cuadrado de Pearson.&#x0D; Resultados: Se registraron 29 casos de mujeres con cáncer de ovario. Once casos (37.9 %) fueron en menores de 45 años. En Etapa clínica III, 18 casos (62.1 %). De grado indiferenciado en 17 casos (58.6 %). 16 casos (55.2 %) fueron del tipo histológico seroso. El valor del CA-125 fue &gt;35 U/mL en 13 casos (61.9 %). Las mujeres con etapa clínica I, tuvieron un Intervalo Libre de la Enfermedad (ILE) de 46 meses versus 27 meses en mujeres con etapa III y 12 meses en etapa IV (P&gt;0.05). El grado de diferenciación celular Bien diferenciado determinó estadísticamente el mejor ILE: 55 meses versus 27 meses en el grado moderadamente diferenciado y de 21 meses en el grado indiferenciado (P=0.025). El tipo histológico así como los niveles de CA-125 no determinaron diferencias estadísticas de ILE, así como en Supervivencia Global Media (SGM). Según el tipo de tratamiento no existieron diferencias estadísticas significativas en el ILE, SGM. Sin embargo las cirugías clasificadas como “óptimas” por parte del equipo quirúrgico tuvieron mayor ILE (Delta 17 meses) P=0.038.&#x0D; Conclusión: En esta serie de casos presentada, las pacientes con cáncer de ovario en Etapa clínica I tuvieron una mejor supervivencia que las pacientes con Etapa clínica III. Así mismo las cirugías clasificadas como óptimas tuvieron mayor intervalo libre enfermedad y mayor supervivencia libre de enfermedad.

NLS-RARα is a novel transcriptional factor.

Acute promyelocytic leukemia (APL) is characterized by the presence of the promyelocytic leukemia (PML)-retinoic acid receptor-α (RAR-α) fusion protein. PML-RARα can be cleaved by neutrophil elastase (NE) in several positions in cells in the promyelocytic stage, nuclear location signal (NLS)-negative PML and NLS-RARα may be the products of PML-RARα by NE. The function of NLS-RARα may be affected by the addition of NLS, which would alter its localization in cells, as the role of NLS is to identify proteins for transport to the nucleus. Preliminary experiments demonstrated that the overexpression of NLS-RARα in HL-60 cells could promote cellular proliferation and inhibit cellular differentiation. Following treatment with all-trans retinoic acid (ATRA), the degree of cellular differentiation was enhanced. In the present study, the localization of NLS-RARα was identified and its activity as a novel transcriptional factor was assessed, which may be critical in the development of APL. The location of NLS-RARα was detected in the nucleus and cytoplasm by indirect immunofluorescence and western blot analysis, with expression in the nucleus revealed to be increased compared with that in the cytoplasm. Next, native-PAGE was performed and NLS-RARα and RXRα were revealed to form heterodimers in the nucleus. In addition, co-immunoprecipitation revealed an interaction between NLS-RARα and retinoid X receptor-α (RXRα). An electrophoresis mobility shift assay (EMSA) indicated that NLS-RARα could bind retinoic acid response elements (RAREs) in the presence of ATRA. Indeed, NLS-RARα could bind RAREs just as WTRARα could, including the RAREs direct repeat-2 (DR-2) and DR-5. In addition, results from a luciferase reporter gene assay demonstrated that NLS-RARα could mediate the activity of RAREs that it bound. Together, these results indicated that NLS-RARα may be a novel transcription factor that contributes to leukemogenesis by competitively binding RAREs as heterodimers with RXRα, just as PML-RARα does, thus repressing the gene transcription essential for myeloid differentiation. These findings indicate the potential role of NLS-RARα targeted therapy in APL.

Abstract 3872: Retinoic acid (RA) relieves EZH2-mediated epigenetic suppression in high-risk neuroblastoma (HR-NB)

Abstract High risk neuroblastoma tumors (HR-NB) are characterized by the presence of undifferentiated histopathology, while tumors from low risk patients have histologic evidence of differentiation and a transcriptome enriched in differentiation genes. Determining the cellular and molecular underpinnings that contribute to this undifferentiated state will lead to better understanding of the disease pathogenesis. Retinoic acid (RA) is known to prime embryonic stem cells for lineage specific differentiation. To elucidate the early events important for reprogramming of NB cells leading to changes in the degree of cellular differentiation, we treated HR-NB cells (KCNR : MYCN amplified) with 5µM RA for 24 hrs. RNA-Seq analysis, to assess early changes in the transcriptome, showed significant changes in gene expression [2fold-up-regulated-446, 2fold-down-regulated-250, FDR≤0.01] which were enriched for biological functions like neuron projection, neuron differentiation, indicating onset of neurogenesis as a marker of differentiation. Gene set enrichment analysis identified significant positive enrichment of NB differentiation (p=0.001) and PRC2 (Polycomb Repressor Complex2) gene sets (p=0.02) along with downregulation of MYCN signature gene set (p=0.001). PRC2 component EZH2 (Histone Methyltransferase) is highly expressed in NB and represses differentiation genes. Though, EZH2 protein levels did not change after treatment, RA caused 40% reduction in H3 bound EZH2. To gain insight into how this reduction affects subsequent gene expression; we evaluated changes in genome wide binding of EZH2 and H3K27me3 after treatment. Indeed, significant correlation was observed between the changes in expression levels of genes after RA treatment and relative binding of EZH2 (p=3.88e-6) and H3K27me3 (p=3.88e-6) at their promoter regions. One-third (n=147) of the upregulated genes (eg. HEY1, RARB) and half (n=117) of the downregulated genes (eg. INSM2, CAMK4) were EZH2 targets. Upregulated genes were related to biological functions like negative regulation of cell growth and transcription, nervous system development. To probe the underlying mechanism, in-silico analysis indicated RA decreased Wnt/ß-catenin, eNOS and PTEN pathways while stimulating cAMP, Integrin linked kinase and G-coupled protein pathways. Evaluation of the cAMP pathway candidates identified 13 genes as EZH2 targets. eg CNR1, DUSP6, PKIA, RSG4. In primary NB tumors, microarray studies indicate that low levels of CNR1 and DUSP6 are associated with poor prognosis. RA stimulates 10-fold and 2-fold increases in CNR1 and DUSP6 levels, respectively. RA treatment inhibits EZH2 binding to its target genes thereby activating pathways involved in reprogramming of HR-NBs to a more differentiated state. Studies are underway to further evaluate the molecular mechanism leading to decreased EZH2 binding upon RA treatment. Citation Format: Deblina Banerjee, Zhihui Liu, Doo-Yi Oh, Maggie Cam, Bong-Hyun Kim, Carol Thiele. Retinoic acid (RA) relieves EZH2-mediated epigenetic suppression in high-risk neuroblastoma (HR-NB) [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2017; 2017 Apr 1-5; Washington, DC. Philadelphia (PA): AACR; Cancer Res 2017;77(13 Suppl):Abstract nr 3872. doi:10.1158/1538-7445.AM2017-3872

Sertoli–Leydig Cell Tumour with Heterologous Element with Rare Mitotic Figure Associated with Multinodular Goitre: A Rare Case of Sex Cord Stromal Tumour of Ovary with Rare Association

Sertoli–Leydig cell tumours are uncommon sex cord stromal tumours of the ovary that often present diagnostic confusion due to the various morphological and histopathological patterns that may be assumed by the tumour. A 28-year-old female presented with abnormal uterine bleeding and thyromegaly is found to have right adnexal mass with elevated serum testosterone. The mass was diagnosed as a Sertoli–Leydig cell tumour, intermediate differentiation with heterologous elements and focal anaplasia. She was treated with surgical resection followed by adjuvant chemotherapy. Definitive diagnosis is made by histopathological study with immunohistochemistry. Prognosis is related to the degree of cellular differentiation and to the presence of heterologous elements with stage of disease at the time of diagnosis.

Hyaluronic Acid in Normal and Neoplastic Colorectal Tissue: Electrospray Ionization Mass Spectrometric and Fluor Metric Analysis

Aim: The aim of the study was to analyze the expression of hyaluronic acid (HA) and its type in human colorectal cancer (CRC) and non-neoplastic mucosa tissues. Methods : The study included 64 adult CRC patients, who met all the inclusion and exclusion criteria. Two tissue samples (neoplastic and non-neoplastic mucosa) from each patient's large intestine were collected and were subjected to electrospray ionization mass spectrometry (ESI-MS) and fluorimetric assays. Results: The analysis of colorectal neoplastic tissue and non-neoplastic mucosa by ESI-MS allowed the identification of HA and its oligosaccharide fragments. Low molecular weight (LMW), nonbiotinylated isoform of HA, and its fragments were identified in both neoplastic and non-neoplastic mucosa. The expression of HA was found to be slightly lower in tumor tissue (0.561 mg HA/g tissue) than in colorectal non-neoplastic mucosa (0.579 mg HA/g tissue), although the difference was not statistically significant ( P = 0.87). This result was probably influenced by the nonbiotinylated LMW-HA. For the specific group of patients that did not present lymph node metastasis, the average HA levels were higher in tumor tissue (0.674 mg HA/g tissue) than in non-neoplastic tissue (0.529 mg HA/g tissue), which managed to reach statistical significance ( P = 0.04). For the group of patients with lymph node involvement, no difference between tumor and nontumor tissue was observed. The HA expression among the tumor tissues within the variables of each clinicopathological parameters assessed failed to elicit any significant difference (location, P = 0.62; size, P = 0.13; lymph node invasion, P = 0.57; degree of cellular differentiation, P = 0.46; venous infiltration, P = 0.73; lymphatic infiltration, P = 0.36; neural infiltration, P = 0.28; tumor node metastasis classification, P = 0.15; and presence of synchronous metastases, P = 0.35; initial versus advanced stage, P = 0.37). Conclusions: The expression of HA was found to be slightly lower in tumor tissue than in colorectal non-neoplastic mucosa, although this difference was not statistically significant. This finding probably influenced the lower expression of HA in tumor tissue than in colorectal non-neoplastic mucosa. Compared to normal tissues, HA levels are significantly increased in the tumor tissues unless they exhibit lymph node metastasis. Otherwise, the expression of HA in tumor tissue did not correlated with the other clinicopathological parameters.

Alpha-fetoprotein Level Predicts Recurrence After Transplantation in Hepatocellular Carcinoma.

Hepatocellular carcinoma (HCC) is one of the leading causes of liver transplantation. In an attempt to predict their recurrence after liver transplantation, evaluation of tumor number and size, degree of histologic differentiation, and the presence of vascular invasion already have their importance established. In this context, the role of biologic markers such as alpha-fetoprotein (AFP) is still not clear. This retrospective cross-sectional study analyzed the AFP relationship with recurrence of HCC after orthotopic liver transplantation.The current study retrospectively analyzed data from 206 patients with a histopathologic confirmed HCC between 1997 and 2010.The overall survival rates at 1, 3, 5, and 14 years were 78.6%, 65.4%, 60.5%, and 38.7%, respectively. The frequency of recurrence was 15.5%, and recurrence was significantly associated with a lower survival rate (P < 0.001). No association was observed between survival and AFP level (P = 0.153). A correlation, however, was found between tumor recurrence and AFP level (P = 0.002). Univariate analysis of risk factors for recurrence revealed that an AFP level greater than 200 ng/mL, the number of tumors, the degree of cellular differentiation, and the presence of vascular invasion or satellite nodules were associated with relapse. By multivariate analysis, only an AFP level greater than 200 ng/mL remained as a risk factor.Although an elevated AFP level did not correlate with survival in HCC patients undergoing orthotopic liver transplantation, a high AFP level was associated with a 3.32-folds increase in the probability of HCC recurrence.

Abstract P2-01-30: A new nomogram to predict axillary metastasis in breast cancer patients without axillary surgery

Abstract Background: Several studies have concentrated on finding a combination of predictive parameters to establish a mathematical model that can identify patients with no axillary metastasis for whom routine lymph node dissection could be safely avoided. We developed a new model of nomogram (the Ulyanovsk Cancer Center axillary lymph node metastasis nomogram, UCC-ALNM nomogram); it employs clinically and pathologically relevant variables and offers possible advantages over the others nomograms. The purpose of the study: To assess the predictive power of UCC-ALNM nomogram. Methods: A total of 530 breast cancer patients treated between 2008 and 2010 were used as the modeling group for validating the UCC-ALNM nomogram. Clinical and pathologic features of patients were assessed by multivariable logistic regression to predict the presence of axillary metastasis in breast cancer patients. The predictive accuracy of our nomogram was measured by calculating the area under the receiver-operating characteristic (ROC) curve (AUC). Clinical factors included into analysis were: patient’s age and localization of the primary tumor. Pathological factors evaluated were: traditional pathological criteria (primary tumor size, histological type, tumor grade, HR- and Her-2 status) and new total pathological index (Ulyanovsk prognostic index - UPI), introduced by pathologists of the Ulyanovsk Regional Cancer Center. UPI is total score of six main pathological criteria that characterize the malignancy of epithelial tumors: degree of cellular differentiation, cellular polymorphism, mitotic activity, growth pattern, lymphovascular invasion, stromal reaction. Results: By the multivariate analysis, patient’s age (p=0.04), tumor size (p&amp;lt;0.001), UPI (p&amp;lt;0.001), PR (p&amp;lt;0.001) and Her2 status (p=0.02) were identified as independent predictors of axillary metastasis. The nomogram was then developed using the six variables associated with axillary metastasis: age, tumor size, PR, Her2, UPI. The new model was accurate and discriminating with an AUC of 0.7510 when applied to the modeling group. Conclusions: UPI is a new predictive factor of axillary metastasis in breast cancer patients. UCC-ALNM nomogram. Citation Format: Valery Rodionov, Vlada Cometova, Sergey Panchenko, Sereda Idrisova, Yurij Savinov, Maria Rodionova. A new nomogram to predict axillary metastasis in breast cancer patients without axillary surgery [abstract]. In: Proceedings of the Thirty-Seventh Annual CTRC-AACR San Antonio Breast Cancer Symposium: 2014 Dec 9-13; San Antonio, TX. Philadelphia (PA): AACR; Cancer Res 2015;75(9 Suppl):Abstract nr P2-01-30.

Structural and functional polymorphism of plastids in leaves of Clivia miniata Rgl. II. Ontogenesis of plastids in mesophyll cells and in cells surrounding vascular bundles

In mesophyll cells and in cells adjoining vascular bundles of leaves in &lt;em&gt;Clivia miniata&lt;/em&gt; Rgl. there occur chloroplasts differing with respect to their structure and the type of accumulated substances. Chloroplasts of mesophyll cells have a well-developed system of grana and intergrana thylakoids; within the stroma they accumulate solely inclusions of lipid character. Chloroplasts of cells surrounding the bundles are smaller than chloroplasts of mesophyll cells, have a poorly developed system of inner membranes and are able to accumulate only considerable amounts of starch. The third type of chloroplasts occurs in parenchymateous cells of the vascular part of the bundles. These chloroplasts, much smaller than those described above, have a reduced system of inner membranes and are not capable of accumulating either starch or lipid droplets. In stroma, they contain only plastoglobules. It seems probable that morphologically uniform proplastids are the precursors of all the studied types of chloroplasts. Their developmental sequence is determined by the degree of cellular differentiation, the type of cell as well as by the function it performs.

Inhibition of LINE-1 retrotransposon-encoded reverse transcriptase modulates the expression of cell differentiation genes in breast cancer cells

Long Interspersed Elements (L1 elements) are biologically active retrotransposons that are capable of autonomous replication using their own reverse transcriptase (RT) enzyme. Expression of the normally repressed RT has been implicated in cancer cell growth. However, at present, little is known about the expression of L1-encoded RT activity or the molecular changes that are associated with RT activity in the development of breast cancer. Here, we report that RT activity is widespread in breast cancer cells. The expression of RT protein decreased markedly in breast cancer cells after treatment with the antiretroviral drug, efavirenz. While the majority of cells showed a significant reduction in proliferation, inhibition of RT was also accompanied by cell-specific differences in morphology. MCF7 cells displayed elongated microtubule extensions that adhered tightly to their substrate, while a large fraction of the T47D cells that we studied formed long filopodia projections. These morphological changes were reversible upon cessation of RT inhibition, confirming their dependence on RT activity. We also carried out gene expression profiling with microarrays and determined the genes that were differentially expressed during the process of cellular differentiation. Genes involved in proliferation, cell migration, and invasive activity were repressed in RT-inhibited cells. Concomitantly, genes involved in cell projection, formation of vacuolar membranes, and cell-to-cell junctions were significantly upregulated in RT-inhibited cells. qRT-PCR examination of the mRNA expression of these genes in additional cell lines yielded close correlation between their differential expression and the degree of cellular differentiation. Our study demonstrates that the inhibition of L1-encoded RT can reduce the rate of proliferation and promote differentiation of breast cancer cells. Together, these results provide a direct functional link between the expression of L1 retrotransposons and the development of breast cancer.

Gd-EOB-DTPA-enhanced 3.0-Tesla MRI findings for the preoperative detection of focal liver lesions: Comparison with iodine-enhanced multi-detector computed tomography

The safety of gadolinium-ethoxybenzyl-diethylenetriamine-pentaacetic-acid (Gd-EOB-DTPA) has been confirmed, but more study is needed to assess the diagnostic accuracy of Gd-EOB-DTPA-enhanced magnetic resonance imaging (MRI) in patients with a hepatocellular carcinoma (HCC) for whom surgical treatment is considered or with a metastatic hepatoma. Research is also needed to examine the rate of detection of hepatic lesions compared to multi-detector computed tomography (MDCT), which is used most frequently to localize and characterize a HCC. Gd-EOB-DTPA-enhanced MRI and iodine-enhanced MDCT imaging were compared for the preoperative detection of focal liver lesions. The clinical usefulness of each method was examined. The current study enrolled 79 patients with focal liver lesions who preoperatively underwent MRI and MDCT. In these patients, there was less than one month between the two diagnostic modalities. Imaging data were taken before and after contrast enhancement in both methods. To evaluate the images, we analyzed the signal-to-noise ratio (SNR) and the contrast-to-noise ratio (CNR) in the lesions and the liver parenchyma. To compare the sensitivity of the two methods, we performed a quantitative analysis of the percentage signal intensity of the liver (PSIL) on a high resolution picture archiving and communication system (PACS) monitor (paired-samples t-test, p < 0.05). The enhancement was evaluated based on a consensus of four observers. The enhancement pattern and the morphological features during the arterial and the delayed phases were correlated between the Gd-EOB-DTPA-enhanced MRI findings and the iodine-enhanced MDCT by using an adjusted x2 test. The SNRs, CNRs, and PSIL all had a greater detection rate in Gd-EOB-DTPA enhanced MRI than in iodine-enhanced MDCT. Hepatocyte-selective uptake was observed 20 minutes after the injection in the focal nodular hyperplasia (FNH, 9/9), adenoma (9/10), and highly-differentiated HCC (grade G1, 27/30). Rim enhancement was detected in all metastases (30/30). During the arterial and the delayed phases, good overall agreement between the gadoxetic-acid-enhanced MR and CT was observed (x2 test, p < 0.05). For the preoperative detection of focal liver lesions, Gd-EOB-DTPA-enhanced MRI had a higher diagnostic value and higher detection rate than iodine-enhanced MDCT. The arterial and the delayed dynamic enhancement patterns, and the gadoxetic-acid-enhanced MR imaging can provide information on the possible degree of cellular differentiation of a HCC, adenoma or metastatic tumor.

56P Predictors of Axillary Lymph Node Metastases in Women with Breast Cancer

ABSTRACT Purpose To assess the impact of different clinical and pathological factors on axillary lymph node involvement in breast cancer patients. Methods A retrospective study of 163 breast cancer patients treated in Ulyanovsk Regional Oncology Center between 2008 and 2010. All the patients received surgery (BCS/mastectomy + axillary lymph node dissection). Clinical factors assessed were: patient's age and tumor localization. Pathological factors: traditional pathological criteria (tumor size, histological variant, grade, HR- and Her-2/neu status) and new total pathological index (Ulyanovsk prognostic index - UPI), introduced by pathologists of our center. UPI is total score of six pathological criteria: degree of cellular differentiation, cellular polymorphism, mitotic activity, growth pattern, LVI, stromal reaction. In accordance with the total score all carcinomas were divided into four groups: 1.Carcinomas of very low malignant potential (UPI scores of 4-6); 2.Low malignant potential (7-10); 3.Moderate malignant potential (11-15); 4.High malignant potential (16-20). Results The median age of the patients was 58.4 ± 11.4 years and the median tumor size was 2.6 ± 1.1 cm. Most of the carcinomas were ER+ (69.3%) and PR+ (65.0%) and Her-2/neu - (82.8%). Of the total 163 breast cancer patients 70 (42.9%) were node positive. Univariative regression analysis revealed that only tumor size (p = 0.03) and UPI (p = 0.00004) are the most powerful predictors of axillary lymph node involvement. There was a lower, but significant association (p = 0.06) between tumor grade and nodal status. Patient's age (p = 0.12), tumor localization (p = 0.79), tumor histology (p = 0.18), ER (p = 0.63), PR (p = 0.52), Her-2/neu status (p = 0.43) had no statistically association with axillary lymph node metastases. Conclusions Tumor size and Ulyanovsk prognostic index are independent prognostic factors for the presence of node metastases. UPI has the highest predictive value, defining malignant potential of the tumor. Disclosure All authors have declared no conflicts of interest.

Serum seleno-proteins status for colorectal cancer screening explored by data mining techniques - a multidisciplinary pilot study

In this case–control pilot study a recent method based on HPLC hyphenated to ICP-MS was employed for the quantification of serum glutathione peroxidase type 3 (GPx3), seleno-protein P (SelP) and seleno-albumin (SeAlb) in 42 patients with colorectal cancer (CRC) and 20 controls. Patients with early cancer stage (TNM I) showed a significantly higher level of SeAlb (19±3ng/mL) in respect to both metastatic CRC patients (TNM IV, 16±4ng/mL) and healthy controls (16±3ng/mL). Classification models based on logistic regression analysis, classification trees and artificial neural networks were constructed using seleno-proteins concentrations as predictors. Neural networks lead to the best performances, up to 95% of corrected predictions in TNM I vs. controls discrimination. These results suggest a potential association between individual seleno-proteins and CRC progression. Age and radiochemoteraphy were assessed as confounding factors, showing no significant effects. Still, SeAlb level tended to reduce with the age in healthy persons, but did not in CRC patients. Seleno-proteins concentration was also compared with a number of clinical parameters considered as prognostic factors in CRC. Significant Spearman's correlations were revealed between SelP and SeAlb, and presence of peritumoural lymphocytic infiltration (ρ=−0.57 and ρ=−0.37, respectively); and SeAlb and degree of cellular differentiation (grading, ρ=−0.37). This study marks the importance to systematically introduce speciation analysis and multidisciplinary approaches in the investigation of the role of seleno-proteins as a potential combined biomarker for CRC.

Expressão da p53 no tumor e no epitélio oral em pacientes com câncer de boca e faringe

INTRODUÇÃO: Expressiva porcentagem de pacientes com carcinomas de boca e faringe apresentam superexpressão da proteína p53 induzida por tabaco, álcool e radioterapia. OBJETIVO: Descrever a expressão da p53 em áreas de mucosa normal adjacente ao tumor e em carcinomas da boca e faringe. MÉTODO: Estudo prospectivo, com seguimento clínico por um ano, de 24 pacientes com câncer espinocelular de boca e faringe. Foram feitas biópsias na neoplasia e em áreas de mucosa normal adjacente ao tumor, antes e 9 meses após a radioterapia, e realizado estudo imunohistoquímico da expressão da p53. RESULTADOS: Antes da radioterapia, houve alteração da expressão da p53 em 20 das 24 biópsias feitas na neoplasia e em 14 nas de mucosa normal adjacente ao tumor. Onze paciente morreram antes de 1 ano de seguimento clínico. Dos 2 pacientes iniciais com aumento da p53 após a radioterapia continuava aumentada em 7 na área da neoplasia e em 6 nas áreas de mucosa normal. Observou-se associação da p53 com o tabagismo e estádio do tumor (p < 5%) mas não com o grau de diferenciação celular e alcoolismo. CONCLUSÃO: O aumento da expressão da p53 foi observado tanto na área da neoplasia como em mucosa normal na maioria dos pacientes com carcinoma de boca e faringe antes e após a radioterapia. Houve correlação estatisticamente significante da expressão da p53 com o tabagismo e estádio da neoplasia.

Differences in the Microrheology of Human Embryonic Stem Cells and Human Induced Pluripotent Stem Cells

Embryonic and adult fibroblasts can be returned to pluripotency by the expression of reprogramming genes. Multiple lines of evidence suggest that these human induced pluripotent stem (hiPS) cells and human embryonic stem (hES) cells are behaviorally, karyotypically, and morphologically similar. Here we sought to determine whether the physical properties of hiPS cells, including their micromechanical properties, are different from those of hES cells. To this end, we use the method of particle tracking microrheology to compare the viscoelastic properties of the cytoplasm of hES cells, hiPS cells, and the terminally differentiated parental human fibroblasts from which our hiPS cells are derived. Our results indicate that although the cytoplasm of parental fibroblasts is both viscous and elastic, the cytoplasm of hiPS cells does not exhibit any measurable elasticity and is purely viscous over a wide range of timescales. The viscous phenotype of hiPS cells is recapitulated in parental cells with disassembled actin filament network. The cytoplasm of hES cells is predominantly viscous but contains subcellular regions that are also elastic. This study supports the hypothesis that intracellular elasticity correlates with the degree of cellular differentiation and reveals significant differences in the mechanical properties of hiPS cells and hES cells. Because mechanical stimuli have been shown to mediate the precise fate of differentiating stem cells, our results support the concept that stem cell “softness” is a key feature of force-mediated differentiation of stem cells and suggest there may be subtle functional differences between force-mediated differentiation of hiPS cells and hES cells.