Arsenic trioxide downregulates cancer procoagulant activity in MCF-7 and WM-115 cell lines in vitro

© 2015 Termedia Sp. z o. o. This is an Open Access article distributed under the terms of the Creative Commons Attribution-NonCommercial-ShareAlike 4.0 International (CC BY-NC-SA 4.0) License (http://creativecommons.org/licenses/by-nc-sa/4.0/), allowing third parties to copy and redistribute the material in any medium or format and to remix, transform, and build upon the material, provided the original work is properly cited and states its license.THE AIM OF THE STUDY: To analyze human breast cancer cell line MCF-7 and human malignant melanoma cell line WM-115 in order to characterize the cellular expression of CP and to evaluate whether ATO may affect this activity, as well as the viability of the cells.MATERIAL AND METHODS: The inhibitory effect of arsenic trioxide on the proliferation of MCF-7 and WM-115 cells were measured with MTT test. The activity of cancer procoagulant after ATO exposure was determined by a specific three-stage chromogenic assay.RESULTS: ATO decreased the CP activity in a dose- and time-dependent manner in MCF-7 cells with no effect on cell proliferation at the same time. However, it affected the CP activity of WM-115 cells in a different way. Reduction in CP activity was followed by an increase after 48 h incubation. The cells viability results showed dose-and time-correlated response within high arsenic concentrations.CONCLUSIONS: Arsenic trioxide downregulates the CP expression in human breast cancer and melanoma cells.Peer reviewedFinal Published versio

Uptake of radiolabelled modified fragment of human alfa-fetoptrotein by experimental mammary adenocarcinoma: in vitro and in vivo studies

BACKGROUND: The aim of the study was to examine in vitro and in vivo binding of radiolabelled analogues of P149 peptide by experimental mammary adenocarcinoma with the intention of potential application for diagnosis and internal radiotherapy of tumours. MATERIAL AND METHODS: The 36-amino acid peptide (P149-QY) of 90% homology to 447–480 peptide fragment of hAFP was synthesised and radiolabelled with iodine-125. The biodistribution of P149-Q[125I]-Y was studied in experimental mammary tumours. For in vitro experiments, extract from mouse mammary tumours were prepared and incubated with radioiodinated P149-QY peptide in the presence of a cross-linking reagent. RESULTS: The gel electrophoresis analysis (SDS-PAGE) showed that radioiodinated P149-QY peptide formed a complex with adenocarcinoma proteins of about 30 kDa. The biodistribution of P149-Q[125I]-Y studied in experimental mammary tumours revealed a higher pharmacokinetic rate in comparison with the whole radioiodinated AFP molecule. A moderate uptake of P149-Q[125I]-Y in the tumour tissue was observed (3.2% ID/g at 30-min p.i.v). However, a faster radioactivity clearance from blood and normal tissues resulted in an increase in the tumour/muscle (T/M) ratio, i.e. from 2.3 to 3.4 after 30 mins and 24 h p.i.v, respectively. CONCLUSIONS: The present study shows that radioiodinated P149-QY peptide reveals some positive features as the AFP receptor radioligand, however, some additional structural modifications of the initial peptide molecule are necessary for full retention of the ligand-receptor interaction of its radiolabelled forms

Estimation of sodium/iodide symporter gene expression (NIS) in thyroid cancer by RT-PCR technique (preliminary study)

Gen NIS zlokalizowany na chromosomie 19 koduje białko złożone z 643 aminokwasów. Białko to należy do rodziny białek błonowych tzw. zależnych od Na+ symporterów glukozowych. W prawidłowej tarczycy jest zlokalizowane w błonie podstawnej tyreocytu. Fizjologiczna rola białka NIS polega na współudziale w biosyntezie hormonów tarczycy, umożliwia bowiem nagromadzanie w znacznym stężeniu jonów I- w tyreocycie. Z doniesień literaturowych wynika, że w nowotworach złośliwych tarczycy (głównie pęcherzykowych i brodawkowatych) ekspresja genu NIS może wpływać na skuteczność rutynowo stosowanej terapii jodem radioaktywnym. Celem pracy było określenie ekspresji genu NIS w rakach tarczycy, a także poszukiwanie zależności pomiędzy ekspresją badanego genu a stopniem zaawansowania klinicznego nowotworu oraz odpowiedzią na zastosowaną radioterapię. Materiał do badań stanowiły mrożone skrawki tkankowe pochodzące z raków tarczycy. Metoda którą wykorzystano do badań była oparta na reakcji odwrotnej transkrypcji (RT) wyizolowanego RNA, po której następowała reakcja PCR wykorzystująca startery swoiste dla genu NIS. Na podstawie uzyskanych wyników stwierdzono, że ekspresja genu NIS pojawia się w różnych typach histologicznych raków tarczycy, głównie rakach brodawkowatych. Nie stwierdzono istotnych statystycznie zależności pomiędzy obecnością ekspresji genu NIS, a stopniem zaawansowania klinicznego zbadanych nowotworów wg klasyfikacji TNM. Nie znaleziono istotnych statystycznie zależności pomiędzy ekspresją badanego genu, a płcią i wiekiem pacjenta. Na bazie dotychczasowych wyników nie ustalono związku pomiędzy ekspresją genu NIS, a jodochwytnością zbadanych przypadków. Ustalenie zależności między ekspresją badanego genu, a skutecznością jodoterapii wymaga dalszych badań, z wykorzystaniem metody ilościowej.NIS gene is located on chromosome 19 and encodes 643 amino acid protein. It belongs to membrane Na+ dependent glucose symporter proteins family. In normal thyroid is located in basolateral membrane of thyreocyte. It plays a main role in concentrating of iodine in thyreocyte and thus in thyroid hormones synthesis. It was proved that NIS expression influences effectiveness of radioactive iodine therapy in well-differentiated thyroid cancers. The aim of this study was to estimate the NIS expression and its dependence with gender, age and stage in thyroid papillary and follicular cancers. The frozen sections of tissue were used as a source of tumor RNA. RT-PCR technique was employed for NIS expression analysis. We did not find dependence between the presence of NIS expression in investigated thyroid cancers and stage of disease estimated according to TNM classification. We also did not find dependence between NIS expression and gender or sex of the patients. Our results suggest that there is no dependence between NIS expression and iodine uptake

POTENTIAL GENETIC AGENT BFL1 FOR TARGETED THERAPY IN CHRONIC LYMPHOCYTIC LEUKEMIA

Background: Many prognostic factors have been identified in chronic lymphocytic leukemia (CLL) but new ones are still desired. The biological characterization of CLL is now being translated into novel treatment strategies. One new prognostic factor, and therapeutic target, may be BFL1. It is both a serum and a molecular marker that contributes to the progression of CLL and its resistance to chemotherapy. The aim of this study was to evaluate the prognostic value of BFL1 and to assess its correlation with other known prognostic markers in CLL for the cladribine and cyclophosphamide regimen (CC). Methods: qPCR TaqMan® Low Density Array was used for gene expression measurements. Assessment of CD38, ZAP70 and BFL-1 proteins expression was done by means of flow cytometry. Serum TK activity was measured by immunoassay. Results: Protein BFL1 expression was found to be significantly higher in CLL patients than healthy volunteers (p=0.001). Moreover its level was significantly higher in patients with no response (NR) to CC therapy (p=0.009). The expression of BFL1 was considerably down regulated during CC treatment and BFL1 mRNA levels were inversely correlated with apoptotic response. In addition, protein BFL1 expression was found to be similar to thymidine kinase (TK) concentration regarding treatment response. As far as other markers are concerned, a positive correlation was identified between BFL1 and TK (r=0.52, p=0.01). Conclusions: Our findings suggest that BFL1 contributes to chemoresistance and may be a co-existing prognostic factor in CLL in the future

Metalloproteinases – importance in cancer development, diagnostics and therapy

Metalloproteinases (MMP) are proteolytic enzymes whose activity is determined by the presence of zinc ion in the active site and are a widely distributed family of protease to ensure the preservation of homeostasis in animals. In human body they also play many important functions participating in the process of embryogenesis as well as in the formation of blood cells or bone formation. The main role, as it seemed at the beginning research on this group of enzymes, was to digest the extracellular matrix. However, over the years, it turned out that metalloproteinases are important part in the regulation of cell biology, in particular a tumor cell. Studies have confirmed the participation of metalloproteinases in all stages of the process of carcinogenesis. At the moment, there are numerous reports of available of various MMPs in different tumors. However, two of these MMP–2 and MMP–9 belonging to the gelatinases seem to be the most common. It has been confirmed also that higher levels usually indicate a worse prognosis for the patient. Therefore, they may prove to be a valuable prognostic indication for clinicians. Attention was paid to the potential use of matrix metalloproteinases in targeted therapy. The pharmaceutical industry has long been trying to launch metalloproteinase inhibitors as possible therapeutics in cancer, seeing in them a potential that, however, is limited to the early stages of development. After initial setbacks related primarily to the low selectivity is used at once more modern methods as monoclonal antibodies or liposomal drug delivery system. Thus, giving the possibility of treatment that does not cause so enormous side effects as conventional therapy

The characterization and importance of cysteine proteases in cancer development

Cysteine proteases also known as thiol or sulfhydryl proteases are enzymes responsible for catalyzing the hydrolysis of peptide bonds in proteins. They take part in many physiological and pathological processes. Their abnormal activity can lead to a number of disease states including tumor growth. They are involved in the process of carcinogenesis at multiple levels– they participate in the invasion, transformation, angiogenesis, apoptosis and metastasis. The best characterised cysteine proteases are the lysosomal cathepsins, that belong to the papain family. So far 11 cysteine cathepsins have been identified: B, L, H, S, K, F, V, X, W, O and C. They take part in the activation of many proenzymes and prohormones, MHC–II–mediated antigen presentation, bone remodeling, reproduction and apoptosis. Research on the role of cysteine proteases in carcinogenesis showed elevated expression of mRNA or enzymatic activity of cathepsins in many human cancers, eg. breast, lung, brain, gastrointestinal tract, head and neck and melanoma. Cancer procoagulant also belongs to the group of cysteine proteases, however its structure and functions are still the subject of research for many scientists. Elevated levels of activity and concentrations of cancer procoagulant in the serum and tissues from patients with cancer disease compared to those observed in healthy people, provides the possibility of using this factor in the diagnosis of cancer. Many preclinical studies have shown that achieving a stop proliferation and reducing the metastatic potential of tumor cells is possible with the use of cysteine proteinases inhibitors. That gives great hope for the possibility of their application in anticancer therapy.Proteinazy cysteinowe, nazywane również proteinazami tiolowymi lub sulfhydrolowymi, to enzymy odpowiedzialne za katalizowanie reakcji hydrolizy wiązań peptydowych w białkach. Biorą udział w licznych procesach fizjologicznych oraz patologicznych. Ich nieprawidłowa aktywność może prowadzić do wielu stanów chorobowych, w tym rozwoju nowotworów. Zaangażowane są one w proces kancerogenezy na wielu poziomach – uczestniczą w inwazji, transformacji nowotworowej, angiogenenezie, apoptozie i powstawaniu przerzutów. Najlepiej scharakteryzowanymi proteinazami cysteinowymi są, należące do rodziny papainy, lizosomalne katepsyny. Dotychczas poznano 11 ludzkich katepsyn cysteinowych: B, L, H, S, K, F, V, X, W, O i C. Biorą one udział w aktywacji wielu proenzymów i prohormonów, pośredniczą w prezentacji antygenu MHC–II, przebudowie kości, procesie reprodukcji oraz apoptozie. Badania nad udziałem proteinaz cysteinowych w procesie kancerogenezy wykazały podwyższoną ekspresję mRNA lub aktywność enzymatyczną katepsyn w wielu ludzkich nowotworach np. raku piersi, płuc, mózgu, żołądkowo–jelitowym, głowy, szyi oraz czerniaku. Do grupy proteinaz cysteinowych należy również prokoagulant nowotworowy, którego struktura, jak i pełnione przez niego funkcje są wciąż przedmiotem badań wielu naukowców. Podwyższony poziom aktywności, jak i stężenia antygenu prokoagulanta nowotworowego w surowicy krwi osób z chorobą nowotworową oraz w pooperacyjnym materiale tkankowym w stosunku do wartości obserwowanych u ludzi zdrowych, świadczy o możliwości wykorzystania tego czynnika w diagnostyce onkologicznej. Liczne badania przedkliniczne dowiodły, że zatrzymanie proliferacji oraz obniżenie potencjału metastatycznego komórek nowotworowych jest prawdopodobne przy użyciu inhibitorów proteinaz cysteinowych. Daje to duże nadzieje na możliwość zastosowania ich w terapii przeciwnowotworowej

Special trade law updating

Od dnia wejścia w życie ustawy z dnia 22 czerwca 2001 r. o wykonywaniu działalności gospodarczej w zakresie wytwarzania i obrotu materiałami wybuchowymi, bronią, amunicją oraz wyrobami i technologią o przeznaczeniu wojskowym lub policyjnym, zwanej dalej "ustawą z dnia 22 czerwca 2001 r." miały miejsce znaczące zmiany w jej obszarze prawnym, zarówno w prawie krajowym, jak i europejskim. To właśnie te czynniki miały istotny wpływ na podjęcie prac nad nowelizacją bądź opracowaniem nowego, adekwatnie przystosowanego do współczesnych realiów aktu prawnego. W artykule wyszczególniono między innymi istotne, zarówno krajowe jak i europejskie dokumenty stanowiące bodziec do zmiany dotychczas obowiązującej w tej Ustawie materii. W części zasadniczej opisano zmiany, które wynikają z zapisów nowej ustawy i wpływają na wykonywanie działalności gospodarczej w przedmiotowym obszarze, natomiast w części końcowej podjęto próbę oceny ustawy w kontekście zapisów, które na etapie przygotowywania i opiniowania projektu, jako propozycje nie zostały uwzględnione, a w opinii autorów zasadnym byłoby ich ujęcie.The law from June 22, 2011, later named as the law, about conducting the economic activities for production and trading of explosive materials, weapons, ammunition, and articles and technologies of military or police designation has been affected by many significant legal changes, both in the country and Europe, since the day of becoming effective. These facts were essential for launching a work on an updating or developing a new legislative act matched to current realities. The paper presents above all some important documents of both country and European origin which initiated some changes in currently binding matter of the Law. The main part describes the changes effected by the records of the new law which affect the performance of economic activity in domain of the subject matter, whereas the final part tries to assess the law by considering some wordings which were rejected at the stage of draft preparation and consultancies, but according to authors it would have been reasonable to include them in the law