Targeting of Deregulated Wnt/β-Catenin Signaling by PRI-724 and LGK974 Inhibitors in Germ Cell Tumor Cell Lines

The majority of patients with testicular germ cell tumors (GCTs) can be cured with cisplatin-based chemotherapy. However, for a subset of patients present with cisplatin-refractory disease, which confers a poor prognosis, the treatment options are limited. Novel therapies are therefore urgently needed to improve outcomes in this challenging patient population. It has previously been shown that Wnt/β-catenin signaling is active in GCTs suggesting that its inhibitors LGK974 and PRI-724 may show promise in the management of cisplatin-refractory GCTs. We herein investigated whether LGK-974 and PRI-724 provide a treatment effect in cisplatin-resistant GCT cell lines. Taking a genoproteomic approach and utilizing xenograft models we found the increased level of β-catenin in 2 of 4 cisplatin-resistant (CisR) cell lines (TCam-2 CisR and NCCIT CisR) and the decreased level of β-catenin and cyclin D1 in cisplatin-resistant NTERA-2 CisR cell line. While the effect of treatment with LGK974 was limited or none, the NTERA-2 CisR exhibited the increased sensitivity to PRI-724 in comparison with parental cell line. Furthermore, the pro-apoptotic effect of PRI-724 was documented in all cell lines. Our data strongly suggests that a Wnt/β-catenin signaling is altered in cisplatin-resistant GCT cell lines and the inhibition with PRI-724 is effective in NTERA-2 CisR cells. Further evaluation of Wnt/β-catenin pathway inhibition in GCTs is therefore warranted

DOTACIE V POL'NOHOSPODARSTVE A ICH VPLYV NA VYSLEDOK HOSPODARENIA POL'NOHOSPODARSKYCH PODNIKOV NA SLOVENSKU

The primary goal of the article is to evaluate the impact of Common Agricultural Policy (CAP) on the profitability of agricultural enterprises in Slovakia. We also analyze the development of Return on Equity (ROE) with and without subsidies on a selected sample of 756 agricultural companies that mean 59 % of all agricultural corporate bodies in Slovakia. We work with the database from The Research Institute of Agricultural and Food Economics (RIAFE) for the period 2000-2006. Since 2004 when Slovakia entered the EU the amount of subsidies has risen, and it still rises. However, it still does not have any positive respond in profitability of agricultural enterprises, and ROE without subsidies is developing negatively. How is it possible? A partial answer to this question can be found in the added value and sales of own products and services

VPLYV ZADLZENOSTI POL'NOHOSPODARSKYCH FIRIEM HOSPODARIACICH V ROZNYCH PRIRODNYCH PODMIENKACH NA ICH RENTABILITU

The primary goal of the article is to evaluate the effects of indebtedness on the profitability of agricultural farms located in less favoured areas (LFA), and the farms which are not located in the LFA. For evaluation of this situation there were used selected financial indicators, development of which was accounted in the period 2000-2005. The base of the data vere created, by aggregate data of corporate bodies. Data comes from the VU EPP database. Indebtedness of agricultural farms reaches in average 35 %. In accordance with an average Slovak company, which indebtedness is 75 %, and in accordance with direction value 50 % it is still under the tolerable value. Lower indebtedness of agricultural farms results from specific factors of agricultural basic industry. These factors are land and climatic conditions, seasonality of production, lower profit margin, and long cycle of production, which means distinction between production and sales phase that causes ups and downs in cash flows of the companies. The relationship between profitability and indebtedness shows non-linear development in the period 2000-2005

VYVOJ RENTABILITY, LIKVIDITY A AKTIVITY VO VYBRANOM SUBORE POL'NOHOSPODARSKYCH PODNIKOV V ROKOCH 200-2008

The paper evaluates the development of return on equity, cash ratio, and asset turnover ratio in agriculture during the period 2000-2008. For evaluation and analyzes there was used the database from The Research Institute of Agricultural and Food Economics (RIAFE) and a selected sample of 2 250 agricultural companies, which means 20 % out of all agricultural body corporate in Slovakia. The criterion for the selection was the best 250 enterprises per each year in selected ratio performance. The evaluation of Return on equity shows that also agriculture can be attractive for investors especially after joining the EU. But the fact that only 2 enterprises had during the years 2000-2008 the best ROE performance needs to be taken into account. The range of generally recommended values for cash ratio is 0.2--0.6 or 0.8. Agricultural enterprises suffer from low liquidity; however the results show that there is a difference between the years 2000-2003 and 2004-2008. After entering the EU the cash ratio increased to higher values than generally recommended. Asset turnover ratio on the other hand decreased after entering the EU in 2004

Ancient retroperitoneal schwannoma imitating seminoma recurrence: A case report

A 28-year-old male with bilateral testicular seminoma underwent bilateral orchiectomy and radiation therapy of the retroperitoneum. After 17 years, he had a retroperitoneal tumor detected, which was removed 7 years later at age 52 because of its progressive enlargement. Due to its partially cystic and partially solid structure, the radiologic findings could not exclude the possibility of regressively altered seminoma metastasis. After radical surgical removal of the tumor, the histopathological and immunohistochemical examination of the tumor revealed ancient schwannoma. These tumors, although unusual, might pose a clinical diagnostic challenge with the risk of undesired overtreatment

ALDH1A3 upregulation and spontaneous metastasis formation is associated with acquired chemoresistance in colorectal cancer cells

Abstract Background Efficiency of colorectal carcinoma treatment by chemotherapy is diminished as the resistance develops over time in patients. The same holds true for 5-fluorouracil, the drug used in first line chemotherapy of colorectal carcinoma. Methods Chemoresistant derivative of HT-29 cells was prepared by long-term culturing in increasing concentration of 5-fluorouracil. Cells were characterized by viability assays, flow cytometry, gene expression arrays and kinetic imaging. Immunomagnetic separation was used for isolation of subpopulations positive for cancer stem cells-related surface markers. Aldehyde dehydrogenase expression was attenuated by siRNA. In vivo studies were performed on SCID/bg mice. Results The prepared chemoresistant cell line labeled as HT-29/EGFP/FUR is assigned with different morphology, decreased proliferation rate and 135-fold increased IC50 value for 5-fluorouracil in comparison to parental counterparts HT-29/EGFP. The capability of chemoresistant cells to form tumor xenografts, when injected subcutaneously into SCID/bg mice, was strongly compromised, however, they formed distant metastases in mouse lungs spontaneously. Derived cells preserved their resistance in vitro and in vivo even without the 5-fluorouracil selection pressure. More importantly, they were resistant to cisplatin, oxaliplatin and cyclophosphamide exhibiting high cross-resistance along with alterations in expression of cancer-stem cell markers such as CD133, CD166, CD24, CD26, CXCR4, CD271 and CD274. We also detected increased aldehyde dehydrogenase (ALDH) activity associated with overexpression of specific ALDH isoform 1A3. Its inhibition by siRNA approach partially sensitized cells to various agents, thus linking for the first time the ALDH1A3 and chemoresistance in colorectal cancer. Conclusion Our study demonstrated that acquired chemoresistance goes along with metastatic and migratory phenotype and can be accompanied with increased activity of aldehyde dehydrogenase. We describe here the valuable model to study molecular link between resistance to chemotherapy and metastatic dissemination

Decreased methylation in the SNAI2 and ADAM23 genes associated with de-differentiation and haematogenous dissemination in breast cancers

Abstract Background In breast cancer (BC), deregulation of DNA methylation leads to aberrant expressions and functions of key regulatory genes. In our study, we investigated the relationship between the methylation profiles of genes associated with cancer invasivity and clinico-pathological parameters. In detail, we studied differences in the methylation levels between BC patients with haematogenous and lymphogenous cancer dissemination. Methods We analysed samples of primary tumours (PTs), lymph node metastases (LNMs) and peripheral blood cells (PBCs) from 59 patients with sporadic disseminated BC. Evaluation of the DNA methylation levels of six genes related to invasivity, ADAM23, uPA, CXCL12, TWIST1, SNAI1 and SNAI2, was performed by pyrosequencing. Results Among the cancer-specific methylated genes, we found lower methylation levels of the SNAI2 gene in histologic grade 3 tumours (OR = 0.61; 95% CI, 0.39–0.97; P = 0.038) than in fully or moderately differentiated cancers. We also evaluated the methylation profiles in patients with different cancer cell dissemination statuses (positivity for circulating tumour cells (CTCs) and/or LNMs). We detected the significant association between reduced DNA methylation of ADAM23 in PTs and presence of CTCs in the peripheral blood of patients (OR = 0.45; 95% CI, 0.23–0.90; P = 0.023). Conclusion The relationships between the decreased methylation levels of the SNAI2 and ADAM23 genes and cancer de-differentiation and haematogenous dissemination, respectively, indicate novel functions of those genes in the invasive processes. After experimental validation of the association between the lower values of SNAI2 and ADAM23 methylation and clinical features of aggressive BCs, these methylation profiles could improve the management of metastatic disease

Prognostic Value of Apoptosis-Inducing Factor (AIF) in Germ Cell Tumors

Apoptosis is a strictly regulated process essential for preservation of tissue homeostasis. This study aimed to evaluate expression of apoptosis inducing factor (AIF) in testicular germ cell tumors (GCTs) and to correlate expression patterns with clinicopathological variables. Formalin-fixed and paraffin-embedded specimens of non-neoplastic testicular tissue and GCTs obtained from 216 patients were included in the study. AIF expression was detected by immunohistochemistry, scored by the multiplicative quickscore method (QS). Normal testicular tissue exhibits higher cytoplasmic granular expression of AIF compared to GCTs (mean QS = 12.77 vs. 4.80, p p = 0.048). We observed significantly lower AIF expression in GCTs compared to normal testicular tissue, which is an uncommon finding in malignant tumors. AIF downregulation might represent one of the mechanisms of inhibition of apoptosis and promotion of cell survival in GCTs

βcatenin is a marker of poor clinical characteristics and suppressed immune infiltration in testicular germ cell tumors

Abstract Background WNT/βcatenin (WNTβ) pathway is activated in early stages of embryonic development. We aimed to evaluate the significance of βcatenin in germ cell tumors (GCTs) and explore associations with the inflamed environment. Methods Surgical specimens from 247 patients were analyzed. Βcatenin expression was detected in the tumor tissue by immunohistochemistry and correlated with clinical characteristics, outcome, PD-L1 expression and systemic immune-inflammation index (SII). The Ingenuity Pathway Analysis (IPA) was used to investigate the immune-cell related effects of βcatenin and PD-L1 encoding genes. Results βcatenin was expressed in 86.2% of GCTs. The expression in seminomas was significantly lower compared to all subtypes of non-seminoma (all P  150) was associated with primary mediastinal non-seminoma (P = 0.035), intermediate/poor risk disease (P = 0.033) and high tumor markers (P = 0.035). We observed a positive correlation with the PD-L1 in tumor and an inverse correlation with the SII. IPA uncovered relationships of CTNNB (βcatenin) and CD274 (PD-L1) genes and their effects on differentiation, proliferation and activation of lymphocyte subtypes. Conclusion Herein, we showed that βcatenin is associated with male adult GCT characteristics as well as supressed immune environment