Study of alternatively spliced variants of estrogen receptor alpha in breast cancer cell lines

Filip Lhota: Study of alternatively spliced variants of estrogen receptor alpha in breast cancer cell lines Abstract: Estrogen receptor α (ER-α) is a transcription factor responsible for mediation of the activities of its natural ligand 17-β-estradiol (E2), the hormone that together with progesterone belongs to the key regulators of mammary epithelial as well as breast cancer cells proliferation. Except to the major gene product consisting of all eight coding exons of ER-α, numerous qualitatively and quantitatively different spliced variants originated from primary transcript by activity of alternative splicing is expressed. Despite that some of these spliced variants have been functionally characterized, their precise role on final ER-α cellular activity remains to be elucidated. The functional characterization of individual alternative forms of ER-α and description of its participation on the overall ER-α activity is important for our understanding of their biogenesis and is also critical for the delineation of molecular bases for ER-α regulation during anti cancer chemotherapy. This work aimed to study the influence of alternatively spliced ER-α variants on the growth characteristics of clones constructed from stable mammary tissue cell lines in regulation to cultivation conditions and cellular..

Identification of hereditary alterations predisposing to breast cancer development using "next-gen" sequencing

Summary: Breast cancer (BC) is the most frequent cancer type in female population of Europe. Approximately 5 - 10 % accounts for its hereditary form which is characterized by high penetrance, early onset, risen recurrence risk and development of other cancers. Mutational analyses of high risk patients identify a predisposing mutation in one of the most studied genes (BRCA1, BRCA2, TP53, ATM, CHEK2, NBS1, PALB2) only in less than one third of tested breast cancer patients. Lately, with the use of new methods of next-generation sequencing, a number of other susceptibility or candidate genes were characterized, but the incidence of their pathogenic alteration is often geographically different. A notable proportion of high risk patients from families with hereditary BC can represent carriers of population-specific, or private mutations. Most of the to date identified BC susceptibility genes codes for proteins involved in DNA repair, especially repair of double strand break DNA repair. Nevertheless the mutation analysis was conducted only on a small fraction of these DNA repair genes. We can expect that in the group of yet nontested genes coding for DNA repair proteins a rare, but clinically important genetic alterations predisposing to BC in affected families can be discovered. This work describes a..

Proposal for the corporate design of FTVS UK

Title of study: Proposal for the corporate design of FTVS UK Study aim: An analysis of the present situation in the area of the faculty's visual style and proposals for its amelioration by means of a graphic manual. Method: Analysis of internal and external documents and a semi-structured interview are used in this Master's Thesis. Results: A complete graphic manual of FTVS will be presented as a final proposal for an amelioration of the present state. Key words: company communication, company identity, corporate design, logo, graphic manua

Marketing research of value of the brand Puma

My thesis introduces a problem of value identification concerning the brand Puma in the Czech Republic. The main objective of this thesis is to find out how is Puma perceived by the Czech population as well as how satisfied and familiar is the Czech population with its products. The theoretical part of the thesis will deal with obtaining information and defining terms in field of marketing research, brand value and propagation. The practical part will include data collection and its processing, which will enable this research to present concrete results. The techniques I will use are electronic questionnaire and depth interview. Keywords: brand, promotion, sponsorship, marketing researc

Germline alterations of the <i>CHEK2</i> gene changing the CHK2 protein structure identified in NHL patients and controls with their frequencies and related odds ratios (OR).

<p>^a New alterations</p><p>^b Two alterations of the <i>CHEK2</i> coding sequence were identified in one patient (c.470T>C and c.1259+1G>C); <i>OR</i>–odds ratio; <i>CI</i>–confidence interval.</p><p>Note: The nomenclature of <i>CHEK2</i> alterations was based on NCBI <i>CHEK2</i> Reference Sequences NG_008150.1 (gene) and NM_007194.3 (mRNA).</p><p>Germline alterations of the <i>CHEK2</i> gene changing the CHK2 protein structure identified in NHL patients and controls with their frequencies and related odds ratios (OR).</p

Overall survival (OS; upper panels) and progression-free survival (PFS; lower panels) in all NHL patients (regardless of histology subtype) classified according to the type of <i>CHEK2</i> alterations.

<p>Panels show: <b>A.</b> the influence of all alterations affecting the CHK2 coding sequence (cds; HR_OS = 1.6; 95% CI 0.79–3.24 and HR_PFS = 2.1; 95% CI 1.12–4.05); <b>B.</b> the influence of the I157T mutation (HR_OS = 1.5; 95% CI 0.62–3.70 and HR_PFS = 3.7; 95% CI 1.42–9.43) and <b>C.</b> the influence of the c.319+43dupA variant (HR_OS = 0.8; 95% CI 0.50–1.15 and HR_PFS = 0.6; 95% CI 0.44–0.89).</p

Association of Germline <i>CHEK2</i> Gene Variants with Risk and Prognosis of Non-Hodgkin Lymphoma

<div><p>The checkpoint kinase 2 gene (<i>CHEK2</i>) codes for the CHK2 protein, an important mediator of the DNA damage response pathway. The <i>CHEK2</i> gene has been recognized as a multi-cancer susceptibility gene; however, its role in non-Hodgkin lymphoma (NHL) remains unclear. We performed mutation analysis of the entire <i>CHEK2</i> coding sequence in 340 NHL patients using denaturing high-performance liquid chromatography (DHPLC) and multiplex ligation-dependent probe amplification (MLPA). Identified hereditary variants were genotyped in 445 non-cancer controls. The influence of <i>CHEK2</i> variants on disease risk was statistically evaluated. Identified <i>CHEK2</i> germline variants included four truncating mutations (found in five patients and no control; P = 0.02) and nine missense variants (found in 21 patients and 12 controls; P = 0.02). Carriers of non-synonymous variants had an increased risk of NHL development [odds ratio (OR) 2.86; 95% confidence interval (CI) 1.42–5.79] and an unfavorable prognosis [hazard ratio (HR) of progression-free survival (PFS) 2.1; 95% CI 1.12–4.05]. In contrast, the most frequent intronic variant c.319+43dupA (identified in 22% of patients and 31% of controls) was associated with a decreased NHL risk (OR = 0.62; 95% CI 0.45–0.86), but its positive prognostic effect was limited to NHL patients with diffuse large B-cell lymphoma (DLBCL) treated by conventional chemotherapy without rituximab (HR-PFS 0.4; 94% CI 0.17–0.74). Our results show that germ-line <i>CHEK2</i> mutations affecting protein coding sequence confer a moderately-increased risk of NHL, they are associated with an unfavorable NHL prognosis, and they may represent a valuable predictive biomarker for patients with DLBCL.</p></div

Germline intronic and silent alterations in the <i>CHEK2</i> gene in NHL patients and controls with their frequencies and related odds ratios (OR).

<p>^a New alterations</p><p>^bThe c.319+43dupA alteration also did not show a statistically significant deviation from the Hardy-Weinberg equilibrium in any of the analyzed groups (all p > 0.05).</p><p>Germline intronic and silent alterations in the <i>CHEK2</i> gene in NHL patients and controls with their frequencies and related odds ratios (OR).</p

Overall survival (OS; upper panels) and progression-free survival (PFS; lower panels) in DLBCL patients.

<p>Panels show: <b>A.</b> the influence of all alterations affecting the CHK2 coding sequence (cds; HR_OS = 2.3; 95% CI 0.77–6.97 and HR_PFS = 2.6; 95% CI 0.91–7.44); <b>B.</b> the influence of the I157T mutation (HR_OS = 2.9; 95% CI 0.70–12.00 and HR_PFS = 5.2; 95% CI 1.25–22.16) and <b>C.</b> the influence of the c.319+43dupA variant (HR_OS = 0.6; 95% CI 0.32–0.97 and HR_PFS = 0.5; 95% CI 0.32–0.86).</p

A schematic diagram showing individual coding exons and flanking intronic sequences affected by the identified <i>CHEK2</i> sequence variants.

<p>The most important structural/functional domains of CHK2 kinase are depicted by color bars [SQ/TQ domain (amino acid (aa) 19–69) in blue, FHA domain (aa 112–175) in yellow, and kinase domain (aa 220–486) in violet]. The left-hand side shows synonymous and intronic <i>CHEK2</i> variants (italicized) while the right-hand side shows CHK2 protein structure-altering variants (frame-shift and missense) that were described in the NHL patients group (in red), controls (green) or in both populations (in black).</p