Hyperaktivace signální dráhy ERK (extracellular signal-regulated kinases) indukovaná mutacemi v genu B-Raf je klíčová pro vývoj a progresi melanomu. Naše předběžné výsledky ukazují, že i v dlaždicových karcinomech hlavy a krku (HNSCC) je dráha ERK aberantně aktivována, i když jinými mechanismy. Aktivace ERK se tak ukazuje jako jeden z klíčových onkogenních procesů v melanomu a HNSCC. To umožňuje současně studovat nádory s různým histogenetickým původem, ale s obdobnými onkogenními změnami. V navrhovaném projektu budeme studovat úlohu ERK dráhy v HNSCC a melanomu a srovnáme mechanismy její aktivace u těchto zhoubných onemocnění. K identifikaci mechanismů aktivace použijeme analýzu genové exprese s následnou shlukovací analýzou a to jak u archivních, tak u nově odebraných biopsií nádorů. Souběžně budeme studovat expresi proteinů, jak kvantitativně tak immunohistochemicky, a aktivaci proteinkináz. Stanovené nádorově-specifické vlastnosti budeme korelovat s klinickými výstupy, včetně rezistence na léčbu vemurafenibem, s cílem navrhnout nové prognostické faktory pro HNSCC a melanom.; Hyperactivation of the extracellular signal-regulated kinases (ERK) signaling pathway due to B-Raf mutations is central to melanoma development and progression. Our preliminary data suggest that aberrant ERK pathway activation, although mediated by different mechanisms, is present also in majority of head and neck squamous cell carcinomas (HNSCC). This finding identifies the ERK activation as a key oncogenic event and provides the opportunity to concurrently analyze tumors of different origin but with similar oncogenic alterations. In this proposal, we will determine the role of the pathway in HNSCC and melanoma and compare the modes of its activation in these cancers. We will use biobanked and newly collected tumor samples to perform transcriptome profiling followed by unsupervised clustering analyses. In parallel, we will perform protein expression studies, immunohistochemistry, and kinase activation studies. Obtained tumor-specific characteristics will be related to clinical outcomes including vemurafenib resistance in order to design new prognostic factors for HNSCC and melano...

• MeSH
• dlaždicobuněčné karcinomy hlavy a krku diagnóza   terapie   MeSH
• lidé MeSH
• MAP kinasový signální systém MeSH
• melanom diagnóza   terapie   MeSH
• nádorové biomarkery MeSH
• prognóza MeSH
• shluková analýza MeSH
• stanovení celkové genové exprese MeSH
• Check Tag
• lidé MeSH

• Konspekt
• Patologie. Klinická medicína
• NLK Obory
• onkologie
• NLK Publikační typ
• závěrečné zprávy o řešení grantu AZV MZ ČR

Aberrant regulation of the cell cycle is a typical feature of all forms of cancer. In head and neck squamous cell carcinoma (HNSCC), it is often associated with the overexpression of cyclin D1 (CCND1). However, it remains unclear how CCND1 expression changes between tumor and normal tissues and whether human papillomavirus (HPV) affects differential CCND1 expression. Here, we evaluated the expression of D-type cyclins in a cohort of 94 HNSCC patients of which 82 were subjected to whole genome expression profiling of primary tumors and paired normal mucosa. Comparative analysis of paired samples showed that CCND1 was upregulated in 18% of HNSCC tumors. Counterintuitively, CCND1 was downregulated in 23% of carcinomas, more frequently in HPV-positive samples. There was no correlation between the change in D-type cyclin expression and patient survival. Intriguingly, among the tumors with downregulated CCND1, one-third showed an increase in cyclin D2 (CCND2) expression. On the other hand, one-third of tumors with upregulated CCND1 showed a decrease in CCND2. Collectively, we have shown that CCND1 was frequently downregulated in HNSCC tumors. Furthermore, regardless of the HPV status, our data suggested that a change in CCND1 expression was alleviated by a compensatory change in CCND2 expression.

• Publikační typ
• časopisecké články MeSH

Arthrospira platensis, a blue-green alga, is a popular nutraceutical substance having potent antioxidant properties with potential anti-carcinogenic activities. The aim of our study was to assess the possible anti-angiogenic effects of A platensis in an experimental model of pancreatic cancer. The effects of an A platensis extract were investigated on human pancreatic cancer cells (PA-TU-8902) and immortalized endothelial-like cells (Ea.hy926). PA-TU-8902 pancreatic tumours xenografted to athymic mice were also examined. In vitro migration and invasiveness assays were performed on the tested cells. Multiple angiogenic factors and signalling pathways were analysed in the epithelial, endothelial and cancer cells, and tumour tissue. The A platensis extract exerted inhibitory effects on both migration and invasion of pancreatic cancer as well as endothelial-like cells. Tumours of mice treated with A platensis exhibited much lesser degrees of vascularization as measured by CD31 immunostaining (P = .004). Surprisingly, the VEGF-A mRNA and protein expressions were up-regulated in pancreatic cancer cells. A platensis inhibited ERK activation upstream of Raf and suppressed the expression of ERK-regulated proteins. Treatment of pancreatic cancer with A platensis was associated with suppressive effects on migration and invasiveness with various anti-angiogenic features, which might account for the anticancer effects of this blue-green alga.

• MeSH
• antioxidancia farmakologie   MeSH
• endoteliální buňky účinky léků   MeSH
• inhibitory angiogeneze farmakologie   MeSH
• lidé MeSH
• myši nahé MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory slinivky břišní farmakoterapie   MeSH
• patologická angiogeneze farmakoterapie   MeSH
• pohyb buněk účinky léků   MeSH
• protinádorové látky farmakologie   MeSH
• signální transdukce účinky léků   MeSH
• Spirulina chemie   MeSH
• upregulace účinky léků   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• myši MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The steadily increasing incidence of malignant melanoma (MM) and its aggressive behaviour makes this tumour an attractive cancer research topic. The tumour microenvironment is being increasingly recognised as a key factor in cancer biology, with an impact on proliferation, invasion, angiogenesis and metastatic spread, as well as acquired therapy resistance. Multiple bioactive molecules playing cooperative roles promote the chronic inflammatory milieu in tumours, making inflammation a hallmark of cancer. This specific inflammatory setting is evident in the affected tissue. However, certain mediators can leak into the systemic circulation and affect the whole organism. The present study analysed the complex inflammatory response in the sera of patients with MM of various stages. Multiplexed proteomic analysis (Luminex Corporation) of 31 serum proteins was employed. These targets were observed in immunohistochemical profiles of primary tumours from the same patients. Furthermore, these proteins were analysed in MM cell lines and the principal cell population of the melanoma microenvironment, cancer‑associated fibroblasts. Growth factors such as hepatocyte growth factor, granulocyte‑colony stimulating factor and vascular endothelial growth factor, chemokines RANTES and interleukin (IL)‑8, and cytokines IL‑6, interferon‑α and IL‑1 receptor antagonist significantly differed in these patients compared with the healthy controls. Taken together, the results presented here depict the inflammatory landscape that is altered in melanoma patients, and highlight potentially relevant targets for therapy improvement.

• MeSH
• chemokiny krev   MeSH
• dospělí MeSH
• fibroblasty asociované s nádorem metabolismus   MeSH
• krevní proteiny analýza   MeSH
• lidé středního věku MeSH
• lidé MeSH
• melanom krev   metabolismus   MeSH
• nádorové biomarkery krev   MeSH
• nádorové buněčné linie MeSH
• pilotní projekty MeSH
• prognóza MeSH
• proteomika metody   MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• studie případů a kontrol MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• senioři nad 80 let MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• srovnávací studie MeSH

Interleukin-6 is a multifaceted cytokine, usually reported as a pro-inflammatory molecule. However, certain anti-inflammatory activities were also attributed to IL-6. The levels of IL-6 in serum as well as in other biological fluids are elevated in an age-dependent manner. Notably, it is consistently reported also as a key feature of the senescence-associated secretory phenotype. In the elderly, this cytokine participates in the initiation of catabolism resulting in, e.g. sarcopenia. It can cross the blood-brain barrier, and so it is in causal association with, e.g. depression, bipolar disorder, schizophrenia, and anorexia. In the cancer patient, IL-6 is produced by cancer and stromal cells and actively participates in their crosstalk. IL-6 supports tumour growth and metastasising in terminal patients, and it significantly engages in cancer cachexia (including anorexia) and depression associated with malignancy. The pharmacological treatment impairing IL-6 signalling represents a potential mechanism of anti-tumour therapy targeting cancer growth, metastatic spread, metabolic deterioration and terminal cachexia in patients.

• MeSH
• interleukin-6 fyziologie   MeSH
• lidé MeSH
• nádory imunologie   metabolismus   MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

The world population of adults aged 60 years or more is increasing globally, and this development can impact skin disease morbidity and mortality, as well as being reflected in the health care system organization. There is substantial evidence that the burden from a remarkable number of skin nonmalignant and malignant conditions is greater in the elderly. Dermatologic research and clinical education in dermatology should focus on both challenges and opportunities created by aging. Skin aging due to intrinsic and extrinsic factors can alter significantly epidermal and dermal structure and functions. Dermal aging can be linked to a great number of complications in routine dermatologic conditions, with slow healing as an example of a severe complication in the elderly. This may be attributed to aged dermal fibroblasts modifying the tissue microenvironment via a shift in their soluble factors and extracellular matrix repertoire. This senescence-associated secretory phenotype can explain the particular proclivity of aged skin to develop malignancies.

• MeSH
• fibroblasty MeSH
• hojení ran MeSH
• kožní nemoci etiologie   MeSH
• kůže cytologie   MeSH
• lidé MeSH
• stárnutí kůže * fyziologie   MeSH
• věkové faktory MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Our review compares evolution of cancer in the human body to the origin of new species from a common ancestor organism with respect to the theory of Charles Darwin. Moreover, the functional role of the tumor microenvironment as a selective pressure actively participating in cancer progression is also demonstrated. Evolutionary aspects of tumor growth and invasion from the point of view of modern therapeutic challenges and opportunities in precision personalized medicine are also discussed.

• MeSH
• biologická evoluce * MeSH
• individualizovaná medicína * MeSH
• invazivní růst nádoru genetika   MeSH
• lidé MeSH
• nádorové mikroprostředí genetika   MeSH
• nádory genetika   MeSH
• proliferace buněk genetika   MeSH
• selekce (genetika) genetika   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH

Melanoma represents a malignant disease with steadily increasing incidence. UV-irradiation is a recognized key factor in melanoma initiation. Therefore, the efficient prevention of UV tissue damage bears a critical potential for melanoma prevention. In this study, we tested the effect of UV irradiation of normal keratinocytes and their consequent interaction with normal and cancer-associated fibroblasts isolated from melanoma, respectively. Using this model of UV influenced microenvironment, we measured melanoma cell migration in 3-D collagen gels. These interactions were studied using DNA microarray technology, immunofluorescence staining, single cell electrophoresis assay, viability (dead/life) cell detection methods, and migration analysis. We observed that three 10 mJ/cm2 fractions at equal intervals over 72 h applied on keratinocytes lead to a 50% increase (p < 0.05) in in vitro invasion of melanoma cells. The introduction cancer-associated fibroblasts to such model further significantly stimulated melanoma cells in vitro invasiveness to a higher extent than normal fibroblasts. A panel of candidate gene products responsible for facilitation of melanoma cells invasion was defined with emphasis on IL-6, IL-8, and CXCL-1. In conclusion, this study demonstrates a synergistic effect between cancer microenvironment and UV irradiation in melanoma invasiveness under in vitro condition.

• MeSH
• fibroblasty cytologie   patologie   MeSH
• imunohistochemie MeSH
• invazivní růst nádoru * MeSH
• keratinocyty patologie   účinky záření   MeSH
• kokultivační techniky MeSH
• kultivované buňky MeSH
• lidé MeSH
• melanom patologie   MeSH
• ultrafialové záření * MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

The incidence of malignant melanoma is rapidly increasing and current medicine is offering only limited options for treatment of the advanced disease. For B‑Raf mutated melanomas, treatment with mutation‑specific drug inhibitors may be used. Unfortunately, tumors frequently acquire resistance to the treatment. Tumor microenvironment, namely cancer‑associated fibroblasts, largely influence this acquired resistance. In the present study, fibroblasts were isolated from a patient suffering from acrolentiginous melanoma (Breslow, 4.0 mm; Clark, IV; B‑Raf V600E mutated). The present study focused on the expression of structural and functional markers of fibroblast activation in melanoma‑associated fibroblasts (MAFs; isolated prior to therapy initiation) as well as in autologous control fibroblasts (ACFs) of the same patient isolated during B‑Raf inhibitor therapy, yet before clinical progression of the disease. Analysis of gene transcription was also performed, as well as DNA methylation status analysis at the genomic scale of both isolates. MAFs were positive for smooth muscle actin (SMA), which is a marker of myofibroblasts and the hallmark of cancer stoma. Surprisingly, ACF isolated from the distant uninvolved skin of the same patient also exhibited strong SMA expression. A similar phenotype was also observed in control dermal fibroblasts (CDFs; from different donors) exclusively following stimulation by transforming growth factor (TGF)‑β1. Immunohistochemistry confirmed that melanoma cells potently produce TGF‑β1. Significant differences were also identified in gene transcription and in DNA methylation status at the genomic scale. Upregulation of SMA was observed in ACF cells at the protein and transcriptional levels. The present results support recent experimental findings that tumor microenvironment is driving resistance to B‑Raf inhibition in patients with melanoma. Such an activated microenvironment may be viable for the growth of circulating melanoma cells.

• MeSH
• bodová mutace MeSH
• chemorezistence * MeSH
• fibroblasty asociované s nádorem účinky léků   metabolismus   patologie   MeSH
• lidé MeSH
• melanom farmakoterapie   genetika   patologie   MeSH
• metylace DNA MeSH
• nádorové buňky kultivované MeSH
• nádorové mikroprostředí * MeSH
• nádory kůže farmakoterapie   genetika   patologie   MeSH
• protoonkogenní proteiny B-raf antagonisté a inhibitory   genetika   MeSH
• senioři MeSH
• transkriptom MeSH
• Check Tag
• lidé MeSH
• senioři MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH
• kazuistiky MeSH

Technický rozvoj mikrochirurgie ve 20. století vedl k výraznému snížení mortality a morbidity operační léčby vestibulárních schwannomů. Jedním z hlavních kritérií hodnocení úspěchu operace se stala pooperační funkce lícního nervu. Retrospektivně jsme analyzovali soubor 90 pacientů operovaných v letech 2010–2012 retrosigmoidním-transmeatálním přístupem. Cílem práce bylo zhodnotit faktory mající vliv na pooperační funkci lícního nervu. Kontinuita lícního nervu byla narušena v devíti případech (10 %). V těchto případech byla vždy provedena rekonstrukce. Jako statisticky významné parametry úspěšnosti zachování kontinuity lícního nervu byly stanoveny pozice nervu vůči tumoru, prodloužení a rozprostření nervu do plochy, dále pak cystický charakter nádoru a míra zasahování do vnitřního zvukovodu. Definitivní výborné až dobré funkce (1.–3. stupeň dle House-Brackmanna) dosáhlo 96 % pacientů, u kterých byl lícní nerv zachován. Ve skupině pacientů po rekonstrukci dosáhlo dobré funkce 67 % pacientů. Závěrečný stimulační práh determinoval časný pooperační výsledek. Neprokázali jsme však vztah mezi výsledkem a definitivní funkcí lícního nervu. V současné době se daří zachovat kontinuitu lícního nervu u většiny operovaných pacientů. Je potřeba definovat prognostické faktory, které jsou spojeny se zvýšeným rizikem poškození lícního nervu, a implementovat tyto faktory do rozhodování o vhodné léčebné strategii.

Technological development in microsurgery during the 20th century has led to a significant decline in mortality and morbidity of surgical management of vestibular schwannoma. Postoperative facial nerve function is among the main criteria of success. A retrospective analysis focused on 90 patients undergoing retrosigmoid-transmeatal surgery of vestibular schwannoma during 2010–2012. The aim of the study was to evaluate perioperative factors associated with postoperative facial nerve function. Facial nerve was discontinued in nine cases (10%) and was subsequently reconstructed. The position of the facial nerve to the tumor, morphological changes (e.g. elongation and splaying), cystic component of a tumor and an extent of tumor growth into the fundus of the internal acoustic meatus were among the parameters affecting facial nerve preservation. Definitive excellent to good function (House-Brackmann grade 1–3) was achieved in 96% of patients in the group with preserved facial nerve continuity. There was 67% of patients with good definitive postoperative function in the facial nerve reconstruction group. Stimulation threshold at the end of the surgery brought no significant information about definitive function of the facial nerve but it predicted early postoperative outcome. At present, the facial nerve continuity can be preserved in the vast majority of surgically managed vestibular schwannomas cases. It is crucial to define prognostic factors influencing the facial nerve injury outcome and include them into a decision-making protocol. Equally, it is necessary to define factors leading to unfavorable outcomes of the facial nerve function despite its anatomical preservation.

• MeSH
• lidé MeSH
• mikrochirurgie metody   MeSH
• nervus facialis chirurgie   patologie   MeSH
• pooperační komplikace MeSH
• retrospektivní studie MeSH
• vestibulární schwannom * chirurgie   MeSH
• zákroky plastické chirurgie MeSH
• Check Tag
• lidé MeSH