Due to the constantly increasing epidemiological burden of our population on oncological diseases, nationwide preventive programs for selected types of malignant tumors have been introduced within the Czech health system. The aim of this thesis is to analyze and evaluate these screenings, such as mammographic screening, cervical screening and screening of the colon and rectum. The subject of the analysis is the mortality rates for breast cancer (C50), cervix (C53) and colon and rectum cancer (C18-21) between 1994 and 2015. The main focus is on question of whether the development of the mortality rate for selected neoplasms depends on the degree of coverage rate by a preventive program.

Breast cancer in women of productive age is a huge problem of the current population. Approximately 52% of women, which were diagnosed with breast cancer in 2013, were aged 20-64. The aim of this thesis is to discover the influence of surgical solution of breast cancer on women return to their original profession. There were 68 suitable participants, who received surgical treatment, were aged 20-64, with finished treatment and who went back to their original employment. The results show that amongst manually working women, 60,9% return to their original employment, in administrative 78% return, which, in acceptance region of 0,05 (p = 0,142), is not a statisticaly significant difference. Furthermore, it was not found that age and lenght of treatment are connected (p=0,040). It appears that the time it takes to return to employment is significantly connected to the type of operation (acceptance region 0,05, p=0,046). This research shows how important it is for women after breast cancer treatment to receive timely rehabilitational intervention and to return to employment and how this affects their quality of life. Key words: Breast cancer, occupational therapy, return to work, cancer survivors, quality of life, quantitative research

This diploma thesis deals with the national screening programs for cancer diagnosis. The goal of this thesis is to find a proper way how to evaluate the effectivity of screening programs as well as their influence on the intensity of mortality from certain types of cancer. For the purpose of finding out necessary information were used data related to the diagnosis of colorectal cancer, a diagnosis of cervical cancer and breast cancer in the population of the Czech Republic between 1977 - 2011. This thesis is divided into eight chapters. The first chapter is an introduction to the topic and contains the description of the main goals. The second chapter defines terms that are crucial for this thesis. The third chapter is devoted to data sources and institutions that collect different types of data and health statistics. The next chapter deals with the epidemiology of all described types of cancer and also provide information on risk factors and symptoms of the disease. The fifth chapter looks back at trends in mortality and incidence of the most common malignant tumors in the Czech Republic. The sixth chapter describes planning and implementation of screening processes. The seventh history of screening programs in the Czech Republic. The eighth chapter deals with the rules and regulations of the EU Council and the World Health Organization. The ninth chapter represents the final assessment of Czech screening programs, compared both to the WHO guidelines and the results in the world. The last chapter is including description of mortality and their reaction on screening programs. Text describes even comparison with two other European countries (Germany, France).

Inactivation of tumor supressor gene BRCA1 causes a life-long risk of breast carcinoma development. Genetic screenings of indicated individuals from high-risk families help to identify large number of sequence variants in known predisposing genes. Majority of discovered variants doesn't have clinical significance yet which causes a big problem for diagnostics. Some of these variants are found within regulatory non-coding regions of gene. A part of the clinical classification of variants is their functional characterization. The goal of this thesis was to create a model system for functional characterization of variants in non-coding regions and to verify its function. Model system was based on targeted gene manipulation by co-transfecting CRISPR-Cas9 construct and donor construct that contained a portion of BRCA1 gene sequence with analyzed modifications, into U2 OS cells. The cells have stably integrated DR-GFP system which allows the activity of homologous recombination (HR) to be determined. Monoallelic modifications were induced into U2 OS cells. These modifications were in a Kozak sequence region of BRCA1 gene. Expression level of BRCA1 mRNA was determined by qRT-PCR, which showed the same levels of mRNA in all cells with analyzed alterations. Next, expression level of BRCA1 protein was...

Breast cancer is a common malignant disease affecting millions of women worldwide. Amplification of HER2 oncogene, a tyrosine kinase receptor, in breast cancer allows application of targeted therapy, but approximately one third of patients develop resistance to treatment. Relocalization of HER2 from the plasma membrane into the mitochondria was found and suggested as one of the potential causes of such resistance. Here we document that the function of mitochondrial HER2 is distinct from that of HER2 in the plasma membrane. Mitochondrial HER2 enhances cancer cell energetic metabolism, proliferation and migration in vitro, and tumour formation in vivo in mice correlating with elevated level of ROS signalling. The kinase activity of mitochondrial HER2 is unaffected, therefore I investigated its role in mitochondrial HER2 function. Moderate, endogenous levels of the kinase activity of mitochondrial HER2 drive pro-tumorigenic properties of breast cancer cells, while constitutive kinase activity sensitizes these cells to cell death and attenuates tumour formation in animal models. On the other hand, impairment of kinase activity due to mutation in the ATP binding site of mitochondrial HER2 supports adherence-independent growth in vitro and tumor growth in vivo. We propose that HER2 function in...

BACKGROUND: The inactivation of the tumor suppressor gene BRCA1 is a predisposing factor for a breast/ovarian cancer development. Formation of cancer-specific alternative splicing variants with aberrant biological properties can represent additional mechanism decreasing the overall BRCA1 activity in DNA double strand break (DDSB) repair. In this study, we analyzed BRCA1 alternative splicing variants BRCA114-15 and 17-19 ascertained previously during the screening of high-risk breast cancer individuals. METHODS: We established a stable MCF-7 cell line-based model system for an in vitro analysis of BRCA1 variants. Using this system, we analyzed the impact of BRCA114-15 and 17-19 variants on DNA repair kinetics using comet assay and confocal immunomicroscopy. The capacity of DNA repair was assessed directly by an in vitro NHEJ assay and indirectly by a mitomycin C sensitivity test. The proliferation activities were determined by a clonogenic assay and growth curves. RESULTS: Overexpression of BRCA114-15 and 17-19 increases the endogenous level of DNA damage, slows down the DDSB repair, and decelerates the initial phase of radiation-induced foci formation and prolongs their persistence. Moreover, BRCA114-15 and 17-19 differentially influence the activity of HR and NHEJ and sensitivity of MCF-7 cells to ionizing...

Summary: Background: An increased risk for development of hereditary breast cancer is associated with germline mutations in BRCA1/2 and the influence of NBN mutations is also supposed. The aim of this study is to specify the frequency of recurrent mutations in BRCA1/2 in unselected breast cancer patients and the frequency of most common pathogenic mutations in NBN in Czech republic, to assess current criteria for genetic testing and to consider the addition of NBN to the tested genes. Methods: Screening for recurrent mutations 5382insC and 300T>G in BRCA1 was performed by RFLP, screening for mutations in exon 11 of BRCA1 was performed by PTT, screening for mutations in a selected region of exon 11 of BRCA2 by DHPLC, and screening for mutations in exon 6 of NBN by HRMA. All the mutations were confirmed by direct sequencing. Results: In 679 unselected breast cancer patients 7 carriers of 5382insC, 3 of 300T>G, and 4 of other mutations in BRCA1 were identified. 2 locally prevalent mutations were found in BRCA2. In 730 controls only one 5382insC BRCA1 mutation was identified. Out of 5 NBN mutations found in 600 high-risk patients two were 657del5 and one R215W. A total of 8 NBN mutation carriers were identified among 703 breast cancer patients, 2 of them 657del5 carriers and three R215W carriers. In 915...

Breast cancer is a common malignant disease affecting millions of women worldwide. Amplification of HER2 oncogene, a tyrosine kinase receptor, in breast cancer allows application of targeted therapy, but approximately one third of patients develop resistance to treatment. Relocalization of HER2 from the plasma membrane into the mitochondria was found and suggested as one of the potential causes of such resistance. Here we document that the function of mitochondrial HER2 is distinct from that of HER2 in the plasma membrane. Mitochondrial HER2 enhances cancer cell energetic metabolism, proliferation and migration in vitro, and tumour formation in vivo in mice correlating with elevated level of ROS signalling. The kinase activity of mitochondrial HER2 is unaffected, therefore I investigated its role in mitochondrial HER2 function. Moderate, endogenous levels of the kinase activity of mitochondrial HER2 drive pro-tumorigenic properties of breast cancer cells, while constitutive kinase activity sensitizes these cells to cell death and attenuates tumour formation in animal models. On the other hand, impairment of kinase activity due to mutation in the ATP binding site of mitochondrial HER2 supports adherence-independent growth in vitro and tumor growth in vivo. We propose that HER2 function in...

Apoptosis represents one of the cell death mechanisms which is realized after the application of taxanes in breast cancer cell lines. Apoptosis induction can be principally triggered either by outer or inner pathway. The aim of the diploma thesis is to contribute to the elucidation of role and mechanisms of the inner mitochondrial pathway of apoptosis induction after taxane application (paclitaxel and SB-T-1216) employing a model of breast carcinoma cell lines SK- BR-3 (nonfunctional p53, functional capase-3) and MCF-7 (functional p53, nonfunctional caspase-3). Specifically, we tested the effect of both employed taxanes on mitochondrial membrane potential, ROS level and the expression and localization of proteins regulating inner mitochondrial pathway. Taxane application resulted in mitochondrial membrane dissipation in SK-BR-3 cell line. However, this was not shown in MCF-7 cell line. We found no changes in Bax and Smac/DIABLO expression after taxane application in both tested cell lines. There was a decrease of Bid expression after taxane application in SK-BR-3 line, but not in MCF-7 line. Taxane application did not lead to the translocation of Bax and Bid (tBid) proteins from cytosol to mitochondria in both tested cell lines. Similarly, there was no Smac/DIABLO release from mitochondria to...

Užití biologických materiálů k náhradě tkání v plastické chirurgii ! Abstrakt v angličtině Background: Biological meshes are biomaterials consisted of extracellular matrix and used in surgery particularly for hernia treatment or thoracic wall reconstruction. They are capable of vascularization, that decreases risk of infection, expecially when used in contaminated fields. This study compared the strength of incorporation and biocompatibility of two porcine-derived grafts (cross-linked and non-cross-linked) in a rat hernia model. In addition, we hypothesized that combination of extracellular matrices with autologous mesenchymal stem cells used for hernia repair would result in increased vascularization and increased strength of incorporation. Methods: Standardized 2 x 4 cm fascial defect was created in 42 Wistar rats and repaired with a cross-linked or a non-cross-linked graft either enriched or non-enriched with stem cells. The rats were sacrificed 3, 6 and 12 months later. The strength of incorporation, vascularization, cellular invasion, foreign body reaction and capsule formation were evaluated. Results: Comparison of stem cell enriched and non-enriched groups showed no significant differences in the capsule thickness, foreign body reaction, cellularization or vascularization. In the non-cross-linked...