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Ovarian cancer is one of the most common cancers in women. Due to its nonspecific symptoms, it is often diagnosed late. This reduces the possibility of successful therapy. The main treatments are surgery and chemotherapy. With late detection, complete surgical removal of the tumor is not possible. The use of chemotherapy is limited by side effects and the resistance of cancer. Therefore, new and promising treatments are being sought. There are studies on the usage of hormone therapy in ovarian cancer. Unfortunately, a small number of these studies provide insufficient evidence of effectiveness. Hence, additional targeted therapies in ovarian cancer, and especially in immunotherapy, are of interest. Immunotherapy uses antibodies directed against specific receptors on cells important in the development of cancer. The results of the latest research on targeted methods, collected in this work, are promising.

Malignant cells in chronic lymphocytic leukemia (CLL) show resistance to apoptosis, as well as to chemotherapy, which are related to deletions or mutations of TP53, high expression of MCL1 and BCL2 genes and other abnormalities. Thus, the main goal of the present study was to assess the impact of chlorambucil (CLB) combined with valproic acid (VPA), a known antiepileptic drug and histone deacetylation inhibitor, on apoptosis of the cells isolated from 17 patients with CLL. After incubation with CLB (17.5 µM) and VPA (0.5 mM), percentage of apoptosis, as well as expression of two TP53 target genes (p21 and HDM2) and two genes from Bcl-2 family (BCL2 and MCL1), were tested. As a result, an increased percentage of apoptosis was observed for CLL cells treated with CLB and VPA, and with CLB alone. Under the treatment with the drug combination, the expression of p21 gene was visibly higher than under the treatment with CLB alone. At the same time, the cultures under CLB treatment showed visibly higher expression of BCL2 than the cultures with VPA alone. Thus, the present study strongly suggests further investigations on the CLB and VPA combination in CLL treatment.

Background: Immune modulatory factors like indoleamine 2,3-dioxygenase 1 (IDO1) generating kynurenine (Kyn) and receptor for advanced glycation end-products (RAGE) contribute to endometrial and cancer microenvironment. Using adequate experimental models is needed to learn about the significance of these molecular factors in endometrial biology. In this paper we study IDO1 activity and RAGE expression in the in vitro cultured primary human endometrial cells derived from cancerous and noncancerous tissue. Methods: The generated primary cell cultures from cancer and noncancerous endometrial tissues were characterized using immunofluorescence and Western Blot for expression of endometrial and cancer markers. IDO1 activity was studied by Kyn quantification with High Performance Liquid Chromatography with Diode Array Detector. Results: The primary cultures of endometrial cells were obtained with 80% success rate and no major genetic aberrations. The cells retained in vitro expression of markers (mucin MUC1 and HER2) or immunomodulatory factors (RAGE and IDO1). Increased Kyn secretion was associated with cancer endometrial cell culture in contrast to the control one. Conclusions: Primary endometrial cells express immune modulatory factors RAGE and IDO1 in vitro associated with cancer phenotype of endometrium.

Szumna Klaudia, Piędel Faustyna, Jasielski Patryk, Habaj Kamila, Grosman Sylwia, Filip Agata. Cancer Stem Cells as a new promising approach of efficient oncological treatment - the review of literature. Journal of Education, Health and Sport. 2020;10(9):115‑120. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2020.10.09.013 https://apcz.umk.pl/czasopisma/index.php/JEHS/article/view/JEHS.2020.10.09.013 https://zenodo.org/record/4016679 The journal has had 5 points in Ministry of Science and Higher Education parametric evaluation. § 8. 2) and § 12. 1. 2) 22.02.2019. © The Authors 2020; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 10.08.2020. Revised: 15.08.2020. Accepted: 06.09.2020. Cancer Stem Cells as a new promising approach of efficient oncological treatment - the review of literature Klaudia Szumna1, Faustyna Piędel1, Patryk Jasielski1, Kamila Habaj1, SylwiaGrosman1, Agata Filip2 Student's Association of Cancer Genetics in Department of Cancer Genetics with Cytogenetic Laboratory. Medical University of Lublin, Chodźki Street 19, 20-093 Lublin, Poland. Department of Cancer Genetics with Cytogenetic Laboratory. Medical University of Lublin, Chodźki Street 19, 20-093 Lublin, Poland. Klaudia Szumna; ORCID:0000-0002-0889-2627; klaudiadaria.szumna@gmail.com Faustyna Piędel; ORCID:0000-0002-8280-498X; faustyna.piedel@gmail.com Patryk Jasielski; ORCID: 0000-0002-0958-735X; patryk.jasielski111@gmail.com Sylwia Grosman; ORCID: 0000-0003-2126-6011; s.grosman@op.pl Kamila Habaj; ORCID: 0000-0002-1733-8046; habajkamila@gmail.com Agata Filip; ORCID: 0000-0001-7591-5887; A.filip@umlub.pl Summary: Introduction and purpose: For many years a lot of research projects have been conducted with the aim to discover special cells, which are responsible for a neoplasm development. One of the theories concerns cancer stem cells (CSCs). This theory presumes the existence of original, undifferentiated cancer cells. These cells are capable of transform to all types of cells in neoplasm mass. The purpose of this study was to review the currently available data about the theory of CSCs and especially potential usage of this theory in oncological treatment. Pubmed and Scopus databases from the last 10 years were searched for phrase “cancer stem cells”. A brief description of the state of knowledge: Impaired signaling pathways lead to an occurrence of CSCs which may contribute to the development of neoplasm eventually. It is proved, that gamma secretase inhibitors impede Notch signaling pathway, which leads to a decrease of tumour markers expression and consequently decline in vivo tumour growth. It is believed that epigenetic mechanisms, including DNA methylation, histones alteration and RNA orientation are pivotal regulators of CSCs. It was shown, that histone deacetylases (HDAC) modulate refractoriness on chemotherapy in haematological tumours. It is also evidenced that peculiar designed microRNA directly inhibit CD44 which suppresses prostatic cancer stem cells and its metastases. CSCs have high expression of programmed death-ligand 1 (PD-L1) including breast and colon cancer, which proclaims in favour of immunotherapy aimed on CSCs. Conclusions: Constantly growing knowledge about CSCs biology is creating new opportunities of detection, isolation and design of the therapies aimed on CSCs. Key words: Cancer stem cells, elective treatment, chemotherapy, remission.

Psiuk Dominika, Rocka Agata, Żak Klaudia, Tomczyk Żaklina, Filip Agata. The efficacy of HDAC inhibitors in neoplasm treatment. Journal of Education, Health and Sport. 2020;10(8):244-250. eISSN 2391-8306. DOI http://dx.doi.org/10.12775/JEHS.2020.10.08.028 https://apcz.umk.pl/czasopisma/index.php/JEHS/article/view/JEHS.2020.10.08.028 https://zenodo.org/record/3987520 The journal has had 5 points in Ministry of Science and Higher Education parametric evaluation. § 8. 2) and § 12. 1. 2) 22.02.2019. © The Authors 2020; This article is published with open access at Licensee Open Journal Systems of Nicolaus Copernicus University in Torun, Poland Open Access. This article is distributed under the terms of the Creative Commons Attribution Noncommercial License which permits any noncommercial use, distribution, and reproduction in any medium, provided the original author (s) and source are credited. This is an open access article licensed under the terms of the Creative Commons Attribution Non commercial license Share alike. (http://creativecommons.org/licenses/by-nc-sa/4.0/) which permits unrestricted, non commercial use, distribution and reproduction in any medium, provided the work is properly cited. The authors declare that there is no conflict of interests regarding the publication of this paper. Received: 01.08.2020. Revised: 05.08.2020. Accepted: 17.08.2020. The efficacy of HDAC inhibitors in neoplasm treatment Dominika Psiuk1, Agata Rocka1, Klaudia Żak1, Żaklina Tomczyk1, Agata Filip2 1Student Research Group, Department of Cancer Genetics, Medical University of Lublin, Radziwillowska 11, 20-080 Lublin, Poland, tel. +48 81 4486102 2Head of the Department of Cancer Genetics, Medical University of Lublin, Radziwillowska 11, 20-080 Lublin, Poland, tel. +48 81 4486102 Dominika Psiuk: ORCID:0000-0003-3319-3489; dominika.psiuk@gmail.com Agata Rocka: ORCID:0000-0003-4738-3160; agatarocka2@gmail.com Klaudia Żak: ORCID:0000-0003-2421-2553; zakklaudia3@gmail.com Żaklina Tomczyk: ORCID:0000-0002-0251-0448; zaklinka512@gmail.com Agata Filip: ORCID: 0000-0001-7591-5887; agata.filip@umlub.pl Abstract Introduction and purpose: Histone Deacetylases (HDACs) are associated with epigenetic regulation of gene expression. They are responsible of adequate action of not only histone proteins, but also crucial cell cycle proteins, such as p53, NF‐κB or alpha tubulin. Recent studies have shown the connection between expression of HDAC and carcinogenesis and impact of HDAC inhibitors on cancer therapy. The aim of the study was to review the recent studies on HDAC inhibitors efficiency in the treatment of hematologic neoplasms and solid tumors. It is a study review from 2010 to 2020 using the PubMed database and ClinicalTrials.gov. Description of state of knowledge: Most studies related to HDAC inhibitors impact in hematological neoplasms, such as multiple myeloma, Hodgkin lymphoma, T-cell lymphomas and acute myeloid leukemia. Among studied agents panobinostat, vorinostat and showed efficacy. Moreover, some HDACi demonstrated effectiveness in solid tumors treatment, foremost vorinostat in breast cancer and entinostat in non-small cell lung lymphoma. Summary: Including HDAC inhibitors in tumor treatment could be beneficial for patients, especially those with advanced, relapsed or refractory hematologic neoplasm. However, further studies are necessary to confirm their efficacy and to reveal side effects. Key words: HDAC, inhibitors, cancer, hematology

Inhibitors of histone deacetylases (HDIs), by affecting the process of histone acetylation, lead to changes in chromatin condensation, and in consequence, to changes in the expression of numerous genes responsible for the cell cycle and differentiation. Therefore, they can be effective in the treatment of cancer. The antitumor activity for over 15 HDIs has been confirmed so far, and some of them have been approved in the USA and Europe, mainly in combination with cytostatics or radiotherapy. For several HDIs, large clinical trials are being carried out to estimate their effectiveness, in monotherapy and new combinations. Other synthetic and natural compounds with HDI activity are tested in preclinical studies. HDIs differ in terms of chemical structure, biological activity and specificity in relation to individual HDACs. The compounds active against classic HDACs (Zn2+-dependent metalloproteins) are usually characterized by the presence the Zn2+ binding group, the linker part and the capping group. Taking into account the type of Zn2+ binding group, classic HDIs are classified as short-chain fatty acids, hydroxamic acids, cyclic peptides and benzamides. In turn, the nicotinamide adenine dinucleotide-dependent SIRTs inhibitors are small molecules, mostly nicotinamide and β-naphthol derivatives. The presented paper summarizes the most important information regarding the use of HDIs as anticancer drugs, regarding their diversified chemical structure, activity against HDACs, additional therapeutic properties and side effects. The review was made taking into account the literature from the last five years (2013-2017).

Oxidative stress and systemic inflammation are closely linked with increased risk of cancer development. Tumor necrosis factor alpha (TNF-α) is one of the pro-inflammatory cytokines. Glutathione S-transferases (GSTs) are enzymes involved in oxidative stress handling. Polymorphisms of genes encoding mentioned molecules may potentially influence the risk and the outcome in neoplastic diseases. Multiple myeloma (MM) is a hematological malignancy characterized by clonal, atypical plasma cell proliferation. In the present study we investigated the association of deletion polymorphisms in GSTT1/GSTM1 genes and single nucleotide polymorphisms (SNPs) in the TNF-α gene at positions −308/−238 with the risk and outcome in MM and sensitivity to bortezomib under in vitro conditions. One hundred newly diagnosed MM patients and 100 healthy blood donors were genotyped by means of multiplex PCR (for GSTs) and PCR-RFLP (for TNF-α). In a subgroup of 50 MM patients, bone marrow cells were treated with bortezomib in vitro. Patients with −238GA+AA or GSTT1-null genotypes had 2.0 (p = 0.002) or 2.29 (p = 0.013) fold increased risk of MM. The interaction effects and risk of MM were observed in GSTT1/GSTM1-null (OR = 2.82, p = 0.018), −308/−238GA+AA (OR = 5.63, p < 0.001), as well as in all combinations of −308 with GSTs. The −308/−238GA+AA genotypes in comparison to GG were associated with earlier MM onset−61.14 vs. 66.86 years (p = 0.009) and 61.72 vs. 66.52 years (p = 0.035), respectively. Patients with GSTM1-present had shorter progression-free-survival (15.17 vs. 26.81 months, p = 0.003) and overall-survival (22.79 vs. 34.81 months, p = 0.039) compared with GSTM1-null. We did not observe relationship between response rate and studied polymorphisms. The in vitro study revealed significantly higher number of apoptotic cells at 12 nM of bortezomib in GSTT1-present, GSTM1-null/present, −308GG and −238GG/GA+AA genotypes. Our findings comprise large analysis of studied polymorphisms in MM.

Chronic lymphocytic leukemia (CLL) is one of the most frequent lymphoproliferative diseases. CLL is characterized by unusual heterogeneity, which probably reflects its biological and genetic lack of homogeneity. Clonal chromosome aberrations belong to the most important prognostic and predictive factors in CLL. This research was aimed at observing clonal evolution in CLL at the chromosomal level, and assessing its clinical significance in relation to selected prognostic factors. The study involved 72 untreated patients with CLL. The preliminary investigations using cytogenetic banding analysis (CBA) and FISH were performed at the time of diagnosis, and again after about 24 months to observe clonal changes (clonal evolution). In addition, other parameters were evaluated, i.e., the expression of ZAP-70 kinase, CD38 antigen, and the mutation statuses of IGVH and NOTCH1 genes. Classic prognostic factors, i.e., categorized ZAP70 and CD38 expressions as well as mutations in IGVH and NOTCH1 genes did not influence the course of clonal evolution in the examined group of patients. Clonal evolution was detected in 45.8% of patients by means of CBA, and in 19.4% patients with FISH. Analysis of chromosomal aberrations in the examined group of patients showed that the incidence of 17p deletions and translocations in karyotypes has a negative impact on overall survival. CE was found to be a risk factor for the occurrence of disease progression (OR = 2.22). Our observations indicate that combined CBA and FISH are the most optimal techniques for monitoring clonal evolution in the course of CLL.

Introduction: The insertion (I allele) deletion (D allele) polymorphism of ACE gene (rs4646994) may influence the etiopathogenesis of multiple myeloma (MM). ACE gene is expressed in bone marrow cells and encodes angiotensin converting enzyme (ACE). It converts angiotensin I to active peptide angiotensin II, which stimulates proliferation of hematopoietic stem cells. This suggests possible association of ACE I/D gene polymorphism with MM. The aim of our study was to check possible impact of this polymorphism on risk of development and outcome of MM, as well as, sensitivity to bortezomib in cell cultures derived from MM patients. Objects and Methods: Genomic DNA from 98 newly diagnosed MM patients and 100 healthy blood donors were analyzed by PCR method. Chromosomal aberrations were detected by use of cIg-FISH. In a subgroup of 40 MM patients nucleated bone marrow cells were treated with bortezomib in vitro. Results: The Hardy-Weinberg equilibrium test showed that genotypic frequencies diverged significantly from the equilibrium. The differences between I and D allele frequencies in control and study population were significant (p = 0.046). We observed the association between DD genotype and more than 2-fold risk of MM - OR = 2.69; p < 0.0001. We did not detect any significant differences among studied genotypes regarding clinical and laboratory parameters. Moreover, we did not observe the association between survival of MM patients and I/D genotypes. Bortezomib increased number of apoptotic and necrotic cells, but the only statistically significant differences were observed in the number of viable cells at 1 nM between ID and DD genotypes (p = 0.026). Conclusion: Presented results confirmed the significant relationship between ACE (I/D) polymorphism and risk of MM development. We did not observe the association of ACE I/D polymorphism with disease outcome and bortezomib in vitro sensitivity.

Jedną z grup lekow immunomodulacyjnych, modyfikujących dzialanie ukladu odpornościowego, są przeciwciala monoklonalne. Negatywne receptory, takie jak CTLA-4 i PD-1, obnizają poziom stymulacji komorki, a w opisywanych przypadkach — hamują aktywacje i proliferacje limfocytow T. Fizjologiczną funkcją tych receptorow jest eliminacja autoreaktywnych komorek odpornościowych, ktore nie zostaly zniszczone w mechanizmie tolerancji centralnej, oraz ochrona tkanek organizmu zmienionych zapalnie. Nowotwory, poprzez ekspresje ligandow dla negatywnych receptorow, unikają rozpoznania przez uklad immunologiczny. Dzialanie przeciwcial obejmuje blokowanie receptorow CTLA-4 i PD-1, przywracając limfocytom zdolnośc rozpoznawania i reagowania na antygeny nowotworowe. Dotychczasowe badania przeprowadzone nad lekami anty-CTLA, anty-PD-1 i anty-PD-L1 wykazują umiarkowaną skutecznośc w leczeniu niektorych zaawansowanych nowotworow, czesto opornych na leczenie standardowe, polączoną z trwalą remisją guza. Szczegolnie korzystne okazalo sie lączenie dwoch grup lekow z efektem synergistycznym. Pomimo zachecających wynikow dotychczasowych badan wciąz przed naukowcami stoją wyzwania, takie jak znalezienie obiektywnych markerow szacujących korzyści leczenia danego pacjenta, zmniejszenie liczby dzialan niepoządanych oraz normalizacja cen nowych lekow.

W niniejszej pracy przedstawiono zalezności pomiedzy zmianami w ekspresji poszczegolnych mikroRNA a powsta­waniem i rozwojem wybranych nowotworow. Dokonano przeglądu doniesien na temat uzyteczności badan nad ekspresją mikroRNA, ktore w przyszlości mogą byc wartościowymi i poządanymi przez diagnostow i lekarzy wskaź­nikami prognostycznymi i predykcyjnymi. Mogą sie one stac podwaliną do opracowania nowych metod leczniczych z wykorzystaniem antysensownych miRNA (antagomiry) czy lekow mających na celu kompensacje ilości cząsteczek w przypadku delecji lub uszkodzen genow dla konkretnych mikroRNA. Daloby to potencjalną mozliwośc regulacji ekspresji genow o znaczeniu strategicznym w procesach związanych z powstawaniem i rozwojem nowotworow.

Chronic lymphocytic leukemia (CLL) mainly affects people older than 60 years. Accumulation of morphologically mature but dysfunctional B-lymphocytes in the bone marrow, lymph nodes and peripheral blood is a characteristic feature of this disease. Chromosomal aberrations are observed in lymphocytes of most CLL patients. Typical alterations include deletions of 13q14 and 11q, trisomy 12, and deletions of 17p. Altered expression of the genes located within involved regions, i.e. microRNA15/16 (13q14.3), ATM (11q22-q23) or TP53 (17p13) may be associated with the development and progression of the disease. Cryptic mutations may also contribute to leukemogenesis. Among others, they affect TP53, NOTCH1, SF3B1 and BIRC genes. CLL is a disease with heterogeneous course. There are two clinical forms – indolent and aggressive. The former is characterized by long time to first treatment and demise usually occurs because of coexisting diseases or is associated with leukemia-dependent immunodeficiency. Rapid clinical course and short overall survival, sometimes in spite of appropriate treatment implementation, is typical for aggressive form of CLL. For patients with this form, the moment of treatment initiation and the choice of first-line therapy are especially important, and depend inter alia on prognostic and predictive factors. Established poor prognostic factors in CLL include chromosomal aberrations, i.e., deletion of 17p or 11q, high ZAP-70 kinase expression, mutations/deletions of TP53, and lack of mutation of immunoglobulin heavy chain variable region genes (IgVH). In this paper we tried to point out the importance of some of the prognostic and predictive factors used routinely in the diagnostic management of CLL. Prognostic and predictive potential of microRNA expression level and recently described cryptic changes in the TP53, NOTCH1, SF3B1 and BIRC3 have also been presented.

B-cell chronic lymphocytic leukemia (B-CLL) is one of the most common leukemias of the elderly. To date, although many prognostic factors are known, none are universal or easily accessible thus allowing for the stratification of patients to slow-go and aggressive-course groups. Recent studies have identified new recurrent mutations in CLL cells, including mutation of the gene encoding one of the spliceosome subunits, SF3B1, mutation or rearrangement of NOTCH1, a gene of well-known tumorigenesis association, and disruption of BIRC3, a member of the inhibitors of apoptosis (IAP) family. This article presents the current state-of-the-art findings concerning the prognostic significance of these new alterations, as well as an explanation of the mechanisms underlying their biological impact on CLL lymphocytes.

Conventional cytogenetics is invariably one of the most important methods used in diagnostics of chronic lymphoproliferations. It complements fluorescence in situ hybridization (FISH) and molecular analysis. Presence of particular chromosomal alterations in chronic lymphocytic leukemia enables patients' stratification into appropriate cytogenetic risk groups and influences treatment decisions. In other non-Hodgkin lymphomas cytogenetic analyses are employed also in minimal residual disease assessment.As lymphocytes in chronic lymphoid malignancies are characterized by low proliferation rate in vitro, it is critical to induce their division in the culture properly. Here, we describe methods of lymphocyte isolation from patient's samples, conditions of cell culture, and the most commonly used mitogens for B- and T-lymphocytes in hemato-oncologic analyses.

Ostre białaczki szpikowe (OBS) stanowią niejednorodną grupę chorób, zróżnicowaną pod względem przebiegu, odpowiedzi na terapię oraz zmian cytogenetycznych i molekularnych. Zmiany te mają znaczenie prognostyczne. Korzystnie rokują chorzy z t(8;21), inv(16)/t(16;16) i t(15;17). Chorzy z prawidłowym kariotypem i aberracjami: 6, 8, –Y, t(9;11) i del(12p) należą do grupy rokowania pośredniego. W przypadku chorych ze złożonym kariotypem lub zmianami typu: inv(3)/t(3;3), t(6;9), –5, –7, del(5q), del(7q), rearanżacje 11q23 – rokowanie jest niekorzystne. Do niekorzystnych molekularnych czynników prognostycznych zalicza się: amplifikację [i]C-MYC[/i], amplifikacje i rearanżacje [i]MLL[/i], [i]FLT3-ITD[/i], mutację [i]WT1[/i], nadekspresję BAALC, [i]ERG[/i] i [i]MN1[/i]. Mutacje genów CEBPA i NPM1 uznaje się za korzystne czynniki prognostyczne. Istotne rokowniczo w OBS są także zmiany ekspresji niektórych miRNA.Heterogenność OBS uzasadnia potrzebę prowadzenia badań cytogenetycznych i molekularnych w celu określenia zmian m.in. u chorych z prawidłowym kariotypem w klasycznym badaniu cytogenetycznym. Techniki macierzowe umożliwiają analizę genomowego DNA oraz profilowanie ekspresji genów w poszukiwaniu sygnatur prognostycznych i predykcyjnych.

Systemic sclerosis (SSc) is an autoimmune disorder characterized by the fibrosis of skin, heart, lung, and kidney as well. Excessive activation of fibroblasts is associated with higher expression of Notch1 and/or Notch3 genes. The constitutive expression of NOTCH genes was described in epithelial cells: epidermal keratinocytes, hair follicle cells and sebaceous glands. The NOTCH signalling pathway may be involved in the development of fibrosis, myofibroblast formation and the process of epithelial-mesenchymal transition. Activation of the NOTCH pathway leads to morphological, phenotypic and functional changes in epithelial cells. Furthermore, inhibition of Notch signalling prevent the development of fibrosis in different models, among them, bleomycin-induced fibrosis and in the Task-1 mause model. Molecular mechanisms, including the role of NOTCH signaling pathway, associated with fibrosis in SSc have not been completely recognized.

Nowadays, diagnostics enables detection of cancer-related mutations. It concerns germline mutations as well as de novo mutations. The number of genetics tests available for specific tumors increase in cessantly. BRCA1 and BRCA2 are highpenetrant genes, but still their functions and mechanisms leading to cancer caused by their mutations remain un known. Germline alterations in BRCA1 and BRCA2 sequences are related to hereditary breast cancer syndrome (HBC), hereditary breast and ovarian can cer syndrome (HBOC), hereditary ovar ian cancer syndrome (HOC). Primary and secondary tumors prevention and choice of the best treatment are the benefits of the detection of BRCA2 mutations. Re search on correlation between the tumor phenotype and the type of mutation is continuously performed. This research may have an impact on the development of more effective drugs.

Loss of heterozygosity (LOH) is implicated in the initiation and progression of various human neoplasia, and is observed in both early or in advanced-stage human cancers. The current study was aimed at investigating the frequency of LOH TP53 in human endometrial carcinoma (EC) metastases. LOH was analyzed using 3 intragenic polymorphisms in 38 primary ECs and corresponding metastatic lesions. Allelic loss at intron 1 was detected in 14 out of 38 (37%) primary carcinomas and in 11 out of 38 (29%) metastatic lesions. LOH at intron 1 in primary and metastatic tumors was concomitantly noted in 8 out of 38 (21%) cases. LOH at intron 4 was seen in 46% (17 out of 37) primary ECs and in 35% (13 out of 37) metastatic lesions. LOH at intron 4 in primary tumor/metastasis was concomitantly demonstrated in 27% (10 out of 33) cases. Allelic loss at exon 4 was detected in 5 out of 33 (15%) primary ECs and in one out of 33 (3%) corresponding metastases. Coexistence of LOH TP53 in primary ECs with metastases at intron 1 and intron 4 was observed in three out of 33 (9%) cases. Correlation between allelic loss at intron 1 in primary ECs and corresponding metastases was found (R = 0.475, p = 0.003). Moreover, there was correlation between LOH at intron 1 in metastastic ECs and allelic imbalance at intron 4 in primary uterine tumors (R = 0.416, p = 0.01). There was a relationship between LOH at intron 4 in primary ECs and corresponding metastatic lesions (R = 0.457, p = 0.004). LOH TP53 at intron 4 correlated with the presence of the neoplasm in the uterine cervix (R = 0.319, p = 0.049), and with the non-endometrioid type of primary tumor (R = 0.371, p = 0.024). There was a significant correlation between exon 4 LOH and patient age (less or equal to 50 years and above this age; R = -0.375, p = 0.032). p53 overexpression was present in thirteen out of 38 (34%) cases, both in primary ECs and in metastatic lesions. Overexpression of p53 was higher in non-endometrioid ECs (three out of 5; 60%) than in endometrioid-type uterine tumors (ten out of 33; 30.3%; p = 0.315). p53 overexpression correlated with the presence of cancer in the lumen of fallopian tube(s) (R = 0.032, p = 0.046), and with allelic loss at intron 1 in primary ECs (R = 0.599, p = 0.0001). In conclusion, LOH occurs not only in primary uterine tumors but also in corresponding metastases, with the higher incidence being reported at intron 4 of the TP53. A significant link existed between LOH TP53 at intron 1 and p53 overexpression in primary ECs, but not in the corresponding metastatic lesions.

Acute myeloblastic leukemia (AML) is a genetically heterogeneous disorder. In approximately 60 % of cases specific recurrent chromosomal aberrations can be identified by modern cytogenetic methods, which enable risk stratification. In these AML subgroups, molecular diagnostics serve as an additional tool to confirm and complement cytogenetic data. In the remaining 40 % of AML cases, with normal karyotype (NK), as well as in core binding factor leukemias (CBF-AMLs), molecular techniques enable the identification of intragenic aberrations and gene expression markers, which further differentiate prognosis of AML. In addition to molecular risk stratification established at the moment of diagnosis, minimal residual disease (MRD) assessment by means of highly sensitive real-time quantitative polymerase chain reaction (RQ-PCR) provides a basis for therapeutic decisions in the course of disease.

Acute myeloid leukemia (AML) constitutes a group of diseases heterogeneous with regard to clinical course, response to therapy as well as genetic features that contribute to disease pathogenesis, progression, and outcome. Numerous molecular lesions found in AML patients serve as prognostic and predictive factors; some of these markers have been included in the latest WHO classification. The analysis of cryptic genomic changes and alterations of gene expression is particularly important in cytogenetically normal AML patients. NPM1 and biallelic CEBPA mutations are favorable prognostic factors, while MLL and FLT3 duplications, WT1, IDH1/IDH2, KIT, TET2, and DNMT3A mutations, as well as overexpression of BAALC, ERG, or MN1 have an adverse prognostic impact. Aside from aberrations involving structural genes, mutations of microRNAs, copy number alterations, and altered methylation of gene promoter regions also influence the development and progression of AML.

Summary The aim of our study was to estimate the expression of the Bcl-x L gene, a member of Bcl-2 family, in NSCLC patients. A total of 60 consecutive patients diagnosed with NSCLC that underwent chemotherapy prior to surgery were reviewed. Bcl-x L expression was assessed on paraffin sections by in situ hybridization (ISH) and immunohistochemistry (IMH). We observed the presence of mRNA of Bcl-x L gene and its protein product overexpression in most patients (60 and 81.7%, respectively). In material examined no significant correlation was observed between the pattern of Bcl-x L or protein expression and any clinicopathological factors evaluated. The expression of Bcl-x L protein was low (less than 10% positive cells) in 11 patients (median survival time 29 months) as compared to 49 patients with overexpression (median survival time 21.0 months). The difference was not of statistic significance ( p =0.27). In examined group the Bcl-x L mRNA was found in 36 patients, while it was absent in 24 cases. Median survival time was 14.5 and 86.5 months, respectively ( p =0.001). In addition, 19.4% of 5-year survivals were achieved in patients with overexpression and 54.2% in patients with no mRNA present ( p =0.002). The percentage of 5-year survival in patients with protein expression assessed by IMH was 30.6% ( p =0.31). The estimation of Bcl-x L expression on mRNA and protein level was compared by the means of sign test and the significant difference was found ( p =0.009). The inconsistency was related to 35% of cases. In comparison with IMH, ISH technique appeared to be more specific and accurate in assessment of 5-year survival (25 and 65%; 65 and 70%, respectively). The results of our study indicate that Bcl-x L mRNA overexpression may suggest poor prognosis in NSCLC.

Abstract 2360 Poster Board II-337 Introduction: Despite their longevity in vivo, CLL lymphocytes die rapidly when put to in vitro cultures, what proves that the resistance to apoptosis is not an intrinsic feature of leukemic cells, but depends on environmental signals. Recently it was shown that mononuclear cells from peripheral blood of CLL patients differentiate in vitro into large, adherent cells that grow in close contact with CLL lymphocytes. They were termed “nurselike cells” (NLCs), because they support leukemic lymphocyte survival in culture. The presence of the cells morphologically and phenotypically similar to NLCs was demonstrated in peripheral lymphatic organs of CLL patients. It may suggest their role in CLL lymphocytes protection in vivo and, as a consequence, point the new target in CLL treatment. Patients and Methods: The study included the group of 65 previously untreated CLL patients, 24 women and 41 men, aged from 36 to 86 yrs. 12 patients (18%) were diagnosed with stage 0 according to Rai, 15 patients (23%) with stage I, 30 patients (46%) with stage II, 5 patients (8%) with stage III and 3 patients (5%) with stage IV. Peripheral blood lymphocytes ex vivo were examined for CD14, CD38, BCL2 and ZAP70 expression by flow cytometry and for BCL2, SURVIVIN and ZAP70 gene expression by RT-PCR. TP53 gene status was assessed by FISH. Lymphocytes of 20 patients were assayed for apoptosis-related gene expression by means of cDNA macroarrays (Clontech). To generate NLCs, PB leukemic cells were cultured in vitro for 14 days on standard medium (RPMI 1640 with L-glutamine, 15% FCS, antibiotics/antimycotics; cell density 3 × 106/ml) and the outgrowth and number of NLCs was assessed in relation to clinical and hematological parameters. NLCs were identified morphologically and by CD31/VIMENTIN protein expression. Results: In 58 cases (89%) the outgrowth of NLCs was observed, while their number differed in cultures of the cells of different patients: in 49 cultures (84.5%) there were over 20 NLCs/mm2 (up to 52 NLCs/mm2), and in 9 cases (15.5%) less than 20 NLCs/mm2. Positive correlation was shown between NLC number and B2M serum level (p=0.044) and absolute monocyte count (p=0.019). Significantly higher NLC number was observed in case of patients with higher CD14+ cell number (p Conclusions: The number of NLC cells obtained in vitro from PBL of CLL patients correlates with B2M serum level and SURVIVIN gene expression in CLL cells ex vivo. High B2M level is a marker of poor prognosis. SURVIVIN represents a family of IAP (Inhibitor of APoptosis) proteins. While rare in PBL of CLL patients, its expression is typical for proliferating leukemic cells pool in pseudofollicle microenvironment. SURVIVIN inhibits apoptosis by blocking caspase-3 and -7. Considering the protective role of NLC cells towards CLL lymphocytes in vitro, these results altogether with observed tendency to shorter survival of patients generating high NLCs number may prove the presence of supportive mechanisms exerted by NLCs in vivo. Disclosures: No relevant conflicts of interest to declare.