To define signaling pathways that drive FSH- and epidermal growth factor (EGF)-like peptide-induced cumulus expansion and oocyte meiotic resumption, in vitro cultured pig cumulus-oocyte complexes were treated with specific protein kinase inhibitors. We found that FSH-induced maturation of oocytes was blocked in germinal vesicle (GV) stage by protein kinase A (PKA), MAPK14, MAPK3/1, and EGF receptor (EGFR) tyrosine kinase inhibitors (H89, SB203580, U0126, and AG1478 respectively) whereas phosphoinositide-3-kinase/v-akt murine thymoma viral oncogene homolog (PI3K/AKT) inhibitor (LY294002) blocked maturation of oocytes in metaphase I (MI). Amphiregulin (AREG)-induced maturation of oocytes was efficiently blocked in GV by U0126, AG1478, and low concentrations of LY294002; H89, SB203580, and high concentrations of LY294002 allowed the oocytes to undergo breakdown of GV and blocked maturation in MI. Both FSH- and AREG-induced cumulus expansion was incompletely inhibited by H89 and completely inhibited by SB203580, U0126, AG1478, and LY294002. The inhibitors partially or completely inhibited expression of expansion-related genes (HAS2, PTGS2, and TNFAIP6) with two exceptions: H89 inhibited only TNFAIP6 expression and LY294002 increased expression of PTGS2. The results of this study are consistent with the idea that PKA and MAPK14 pathways are essential for FSH-induced transactivation of the EGFR, and synthesis of EGF-like peptides in cumulus cells and MAPK3/1 is involved in regulation of transcriptional and posttranscriptional events in cumulus cells required for meiotic resumption and cumulus expansion. PI3K/AKT signaling is important for regulation of cumulus expansion, AREG-induced meiotic resumption, and oocyte MI/MII transition. The present data also indicate the existence of an FSH-activated and PKA-independent pathway involved in regulation of HAS2 and PTGS2 expression in cumulus cells.

• MeSH
• cyklooxygenasa 2 genetika   MeSH
• erbB receptory fyziologie   MeSH
• folikuly stimulující hormon farmakologie   MeSH
• glukuronosyltransferasa genetika   MeSH
• glykoproteiny farmakologie   MeSH
• inhibitory proteinkinas farmakologie   MeSH
• kumulární buňky fyziologie   MeSH
• meióza účinky léků   fyziologie   MeSH
• mezibuněčné signální peptidy a proteiny farmakologie   MeSH
• mitogenem aktivovaná proteinkinasa 14 antagonisté a inhibitory   fyziologie   MeSH
• oocyty účinky léků   fyziologie   MeSH
• proteinkinasy závislé na cyklickém AMP antagonisté a inhibitory   fyziologie   MeSH
• regulace genové exprese fyziologie   MeSH
• signální transdukce účinky léků   fyziologie   MeSH
• Sus scrofa fyziologie   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The maturation of mammalian oocytes in vitro can be stimulated by gonadotropins (follicle-stimulating hormone, FSH) or their intrafollicular mediator, epidermal growth factor (EGF)-like peptide-amphiregulin (AREG). We have shown previously that in pig cumulus-oocyte complexes (COCs), FSH induces expression and the synthesis of AREG that binds to EGF receptor (EGFR) and activates the mitogen-activated protein kinase 3/1 (MAPK3/1) signaling pathway. However, in this study we found that FSH also caused a rapid activation of MAPK3/1 in the cumulus cells, which cannot be explained by the de novo synthesis of AREG. The rapid MAPK3/1 activation required EGFR tyrosine kinase (TK) activity, was sensitive to SRC proto-oncogene non-receptor tyrosine kinase (SRC)-family and protein kinase C (PKC) inhibitors, and was resistant to inhibitors of protein kinase A (PKA) and metalloproteinases. AREG also induced the rapid activation of MAPK3/1 in cumulus cells, but this activation was only dependent on the EGFR TK activity. We conclude that in cumulus cells, FSH induces a rapid activation of MAPK3/1 by the ligand-independent transactivation of EGFR, requiring SRC and PKC activities. This rapid activation of MAPK3/1 precedes the second mechanism participating in the generation and maintenance of active MAPK3/1-the ligand-dependent activation of EGFR depending on the synthesis of EGF-like peptides.

• MeSH
• aktivace transkripce MeSH
• amfiregulin metabolismus   MeSH
• erbB receptory metabolismus   MeSH
• extracelulárním signálem regulované MAP kinasy genetika   MeSH
• folikuly stimulující hormon farmakologie   MeSH
• IVM techniky MeSH
• kultivované buňky MeSH
• kumulární buňky cytologie   účinky léků   metabolismus   MeSH
• mitogenem aktivovaná proteinkinasa 1 genetika   MeSH
• mitogenem aktivovaná proteinkinasa 3 genetika   MeSH
• oocyty cytologie   účinky léků   metabolismus   MeSH
• prasata MeSH
• signální transdukce účinky léků   MeSH
• skupina kinas odvozených od src-genu metabolismus   MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH

BACKGROUND: The gonadotropin-induced resumption of oocyte meiosis in preovulatory follicles is preceded by expression of epidermal growth factor (EGF)-like peptides, amphiregulin (AREG) and epiregulin (EREG), in mural granulosa and cumulus cells. Both the gonadotropins and the EGF-like peptides possess the capacity to stimulate resumption of oocyte meiosis in vitro via activation of a broad signaling network in cumulus cells. To better understand the rapid genomic actions of gonadotropins (FSH) and EGF-like peptides, we analyzed transcriptomes of cumulus cells at 3 h after their stimulation. METHODS: We hybridized aRNA from cumulus cells to a pig oligonucleotide microarray and compared the transcriptomes of FSH- and AREG/EREG-stimulated cumulus cells with untreated control cells and vice versa. The identified over- and underexpressed genes were subjected to functional genomic analysis according to their molecular and cellular functions. The expression pattern of 50 selected genes with a known or potential function in ovarian development was verified by real-time qRT-PCR. RESULTS: Both FSH and AREG/EREG increased the expression of genes associated with regulation of cell proliferation, cell migration, blood coagulation and extracellular matrix remodeling. FSH alone induced the expression of genes involved in inflammatory response and in the response to reactive oxygen species. Moreover, FSH stimulated the expression of genes closely related to some ovulatory events either exclusively or significantly more than AREG/EREG (AREG, ADAMTS1, HAS2, TNFAIP6, PLAUR, PLAT, and HSD17B7). In contrast to AREG/EREG, FSH also increased the expression of genes coding for key transcription factors (CEBPB, FOS, ID1/3, and NR5A2), which may contribute to the differing expression profiles of FSH- and AREG/EREG-treated cumulus cells. CONCLUSIONS: The impact of FSH on cumulus cell gene transcription was higher than the impact of EGF-like factors in terms of the number of cell functions affected as well as the number of over- and underexpressed genes. Both FSH and EGF-like factors overexpressed genes involved in the post-ovulatory switch in steroidogenesis and tissue remodelling. However, FSH was remarkably more efficient in the up-regulation of several specific genes essential for ovulation of matured oocytes and also genes that been reported to play an important role in maturation of cumulus-enclosed oocytes in vitro.

• MeSH
• amfiregulin farmakologie   fyziologie   MeSH
• epidermální růstový faktor farmakologie   fyziologie   MeSH
• epiregulin farmakologie   fyziologie   MeSH
• folikuly stimulující hormon farmakologie   fyziologie   MeSH
• kultivované buňky MeSH
• kumulární buňky účinky léků   metabolismus   MeSH
• oocyty účinky léků   metabolismus   MeSH
• prasata MeSH
• regulace genové exprese MeSH
• zvířata MeSH
• Check Tag
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The immune microenvironment in inflammatory breast cancer (IBC) is poorly characterised, and molecular and cellular pathways that control accumulation of various immune cells in IBC tissues remain largely unknown. Here, we discovered a novel pathway linking the expression of the tetraspanin protein CD151 in tumour cells with increased accumulation of macrophages in cancerous tissues. It is notable that elevated expression of CD151 and a higher number of tumour-infiltrating macrophages correlated with better patient responses to chemotherapy. Accordingly, CD151-expressing IBC xenografts were characterised by the increased infiltration of macrophages. In vitro migration experiments demonstrated that CD151 stimulates the chemoattractive potential of IBC cells for monocytes via mechanisms involving midkine (a heparin-binding growth factor), integrin α6β1, and production of extracellular vesicles (EVs). Profiling of chemokines secreted by IBC cells demonstrated that CD151 increases production of midkine. Purified midkine specifically stimulated migration of monocytes, but not other immune cells. Further experiments demonstrated that the chemoattractive potential of IBC-derived EVs is blocked by anti-midkine antibodies. These results demonstrate for the first time that changes in the expression of a tetraspanin protein by tumour cells can affect the formation of the immune microenvironment by modulating recruitment of effector cells to cancerous tissues. Therefore, a CD151-midkine pathway can be considered as a novel target for controlled changes of the immune landscape in IBC. © 2020 Pathological Society of Great Britain and Ireland. Published by John Wiley & Sons, Ltd.

• MeSH
• antigeny CD151 imunologie   metabolismus   MeSH
• chemokiny metabolismus   MeSH
• lidé MeSH
• makrofágy metabolismus   patologie   MeSH
• midkin metabolismus   MeSH
• nádorové buněčné linie MeSH
• nádorové mikroprostředí fyziologie   MeSH
• zánětlivé nádory prsu metabolismus   patologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

The role of the local microenvironment in influencing cell behavior is central to both normal development and cancer formation. Here, we show that sprouty 1 (SPRY1) modulates the microenvironment to enable proper mammary branching morphogenesis. This process occurs through negative regulation of epidermal growth factor receptor (EGFR) signaling in mammary stroma. Loss of SPRY1 resulted in up-regulation of EGFR-extracellular signal-regulated kinase (ERK) signaling in response to amphiregulin and transforming growth factor alpha stimulation. Consequently, stromal paracrine signaling and ECM remodeling is augmented, leading to increased epithelial branching in the mutant gland. By contrast, down-regulation of EGFR-ERK signaling due to gain of Sprouty function in the stroma led to stunted epithelial branching. Taken together, our results show that modulation of stromal paracrine signaling and ECM remodeling by SPRY1 regulates mammary epithelial morphogenesis during postnatal development.

• MeSH
• adaptorové proteiny signální transdukční nedostatek   metabolismus   MeSH
• amfiregulin farmakologie   MeSH
• buňky stromatu účinky léků   metabolismus   MeSH
• časosběrné zobrazování MeSH
• epitel růst a vývoj   metabolismus   MeSH
• epitelové buňky cytologie   účinky léků   MeSH
• erbB receptory metabolismus   MeSH
• extracelulární matrix metabolismus   MeSH
• extracelulárním signálem regulované MAP kinasy metabolismus   MeSH
• fibroblasty účinky léků   metabolismus   MeSH
• fosfoproteiny nedostatek   metabolismus   MeSH
• fosforylace účinky léků   MeSH
• kolagen metabolismus   MeSH
• ligandy MeSH
• membránové proteiny nedostatek   metabolismus   MeSH
• mléčné žlázy zvířat účinky léků   metabolismus   MeSH
• morfogeneze * účinky léků   MeSH
• mutace genetika   MeSH
• myši knockoutované MeSH
• myši nahé MeSH
• parakrinní signalizace * účinky léků   MeSH
• pohyb buněk účinky léků   MeSH
• protoonkogenní proteiny c-akt metabolismus   MeSH
• signální transdukce * účinky léků   MeSH
• transformující růstový faktor alfa farmakologie   MeSH
• vývojová regulace genové exprese účinky léků   MeSH
• zvířata MeSH
• Check Tag
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH
• Research Support, N.I.H., Extramural MeSH

Moderní kombinovaná chemoterapie s cílenou anti-EGFR léčbou vede u senzitivních pacientů s metastatickým kolorektálním karcinomem k prodloužení života a zlepšení jeho kvality. U rezistentních pacientů však použití monoklonálních anti-EGFR protilátek může naopak vést ke zhoršení parametrů přežití. Identifikace senzitivních a rezistentních pacientů se tak stala klíčovou otázkou při rozhodování o léčebné strategii. Četné klinické studie vedly ke zjištění, že rezistence k anti-EGFR terapii je v naprosté většině případů spojena s trvalou aktivací signálních drah distálně od EGFR. Z mnoha studovaných faktorů se do rutinní praxe dostaly pouze mutace genů KRAS a NRAS. U ostatních markerů (mutace BRAF, PIK3CA, inaktivace PTEN a TP53, amplifikace EGFR a HER-2, zvýšená hladina ligandů epiregulinu a amphiregulinu, microRNA miR-31-3p, miR-31-5p a další) je třeba dalších studií. Přesnost prediktivní diagnostiky je možno zvýšit kombinací vyšetřovaných markerů např. pomocí metod založených na sekvenování nové generace.

The combination of modern systemic chemotherapy and anti-EGFR monoclonal antibodies improves overall survival and the quality of life for patients with metastatic colorectal cancer. By contrast, the addition of anti-EGFR therapy to the treatment regime of resistant patients may lead to worse progression-free and overall survival. Therofore, identifying sensitive and resistant patients is key during initial decision-making. A number of clinical trials show that primary resistance to EGFR blockade is in most cases caused by constitutive activation of signalling pathways downstream of EGFR. Of the many biomarkers studied, only the KRAS and NRAS mutation status has reached clinical relevance in routine practice. The other markers (BRAF and PIK3CA mutations, PTEN and TP53 inactivation, EGFR and HER-2 amplification, epiregulin and amphiregulin overexpression, microRNA miR-31-3p and miR-31-5p etc.) still need to be validated. The accuracy of predictive diagnostic tools could also be increased by a combination of predictive markers on the next generation sequencing platform.

• MeSH
• amplifikace genu MeSH
• erbB receptory * antagonisté a inhibitory   terapeutické užití   MeSH
• geny ras MeSH
• kolorektální nádory * diagnóza   MeSH
• kombinovaná farmakoterapie MeSH
• lidé MeSH
• metastázy nádorů MeSH
• mutace MeSH
• nádorové biomarkery analýza   MeSH
• prediktivní hodnota testů MeSH
• protoonkogenní proteiny B-raf MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

Background and Objectives: Initial diagnosis of brest cancer (BC) is important for fate and prognosis of the diseases profile, we sought to identify the correlation between Midkine (MK) as a new biomarker with cancer antigen (CA)15-3, liver function test, renal function test, blood cells parameters in individuals with invasive ductal carcinoma.Methods: The serum MK and CA15-3 of all subjects were measured by the ELISA technique, Liver enzymes were measured by colourimetric methods and neutrophils, and lymphocytes were measured by an Electrical Impedance Cell Counting method (automated machine).Results: The results of the correlation among serum MK and other parameters in invasive ductal carcinoma of the breast showed a considerable positive correlation among MK and CA15-3 and measured white blood cells. Moreover, there were a weak correlation with Aspartate Aminotransferase (AST) and RBC, while there is no correlation between serum MK and other liver enzymes or blood parameters. Conclusion: The study results of the correlation between serum MK and other parameters in colorectal carcinoma patients show a significant positive correlation of MK with CA15-3 markers in invasive ductal carcinoma of the breast.

• MeSH
• duktální karcinom prsu krev   MeSH
• ELISA metody   MeSH
• jaterní testy MeSH
• klinická studie jako téma metody   MeSH
• lidé MeSH
• lymfocyty metabolismus   MeSH
• midkin * analýza   krev   MeSH
• nádorové biomarkery * krev   MeSH
• nádory prsu * diagnóza   krev   MeSH
• neutrofily metabolismus   MeSH
• vyšetření funkce ledvin MeSH
• Check Tag
• lidé MeSH

Background: The second-leading source of neoplasm-related death and a primary factor in gastrointestinal cancer, colorectal cancer (CCR) affects both genders globally. Poor eating behaviours, tobacco, an intestinal inflammatory disorder, swellings, inherited characteristics, and the elderly all increase the threat of acquiring this malignancy. The illness is more hostile in patients detected at earlier ages, although 90% of patients with colorectal tumours are older than 50, with a median oldness of 64 years. American Cancer Association estimates that it caused more than 49,700 fatalities in 2015.Objectives: Study the correlation of midkine with carcinoembryonic antigen (CEA), liver function tests, and white blood cell count in patients with colorectal carcinoma.Methods: The serum midkine and CEA of all subjects were measured by the ELISA technique, Liver enzymes were measured by colourimetric methods and neutrophils, and lymphocytes were measured by an Electrical Impedance Cell Counting method (automated machine).Conclusion: The study results of the correlation between serum midkine and other parameters in colorectal carcinoma patients show a significant positive correlation of midkine with CEA, liver enzyme, neutrophils, and lymphocytes.

• MeSH
• dospělí MeSH
• hematologické testy metody   MeSH
• jaterní testy metody   MeSH
• karcinoembryonální antigen krev   MeSH
• kolorektální nádory * krev   MeSH
• leukocyty MeSH
• lidé středního věku MeSH
• lidé MeSH
• midkin * krev   MeSH
• nádorové biomarkery krev   MeSH
• prognóza MeSH
• statistika jako téma MeSH
• Check Tag
• dospělí MeSH
• lidé středního věku MeSH
• lidé MeSH
• mužské pohlaví MeSH
• ženské pohlaví MeSH
• Publikační typ
• klinická studie MeSH

Východiska: Kombinace moderní chemoterapie s cílenou anti-EGFR léčbou vede u senzitivních pacientů s metastatickým kolorektálním karcinomem k prodloužení života a zlepšení jeho kvality. U rezistentních pacientů může však přidání monoklonálních protilátek proti EGFR vést naopak ke zhoršení parametrů přežití. Z tohoto důvodu se identifikace senzitivních a rezistentních pacientů stala klíčovou záležitostí při iniciální rozvaze před zahájením cílené léčby metastatického kolorektálního karcinomu. Četné klinické studie vedly ke zjištění, že rezistence k anti-EGFR terapii je v naprosté většině případů spojena s trvalou aktivací signálních drah distálně od EGFR. Z mnoha studovaných faktorů (mutace onkogenů KRAS, NRAS, BRAF a PIK3CA, inaktivace nádorového supresoru PTEN a TP53, amplifikace EGFR a HER2, zvýšená hladina ligandů epiregulinu a amphiregulinu, mikroRNA miR-31-3p a miR-31-5p a další) se do rutinní klinické praxe dostaly pouze KRAS a NRAS. U ostatních faktorů je třeba dalších studií k verifikaci zjištěných závěrů. Na pokračující efektivitu anti-EGFR terapie mohou ukazovat i některé klinické parametry zjištěné až po zahájení cílené léčby, jako např. časná regrese nádoru, hloubka nádorové odpovědi či míra poklesu plazmatické hladiny hořčíku. Přesnost prediktivní dia­gnostiky lze zvýšit rovněž kombinací vyšetřovaných bio­markerů např. pomocí metod založených na sekvenování nové generace. Je však třeba varovat před nekritickým vyšetřováním řady molekulárních markerů, které může vést k problémům s interpretací získaných výsledků, především jejich klinické relevance. Cíl: Cílem tohoto přehledu je popsat současné možnosti predikce odpovědi na anti-EGFR terapii v kontextu EGFR signální dráhy a návaznosti na běžnou klinickou praxi.

Background: The combination of modern systemic chemotherapy and anti-EGFR monoclonal antibodies improves overall survival and quality of life for patients with metastatic colorecal cancer. By contrast, the addition of anti-EGFR therapy to the treatment regime of resistant patients may lead to worse progression-free survival and overall survival. Therefore, identifying sensitive and resistant patients prior to targeted therapy of metastatic colorecal cancer is a key point during the initial decision making process. Previous research shows that primary resistance to EGFR blockade is in most cases caused by constitutive activation of signaling pathways downstream of EGFR. Of all relevant factors (mutation of KRAS, NRAS, BRAF, and PIK3CA oncogenes, inactivation of tumor suppressors PTEN and TP53, amplification of EGFR and HER2, and expression of epiregulin and amphiregulin, mikroRNA miR-31-3p, and miR-31-5p), only evaluation of KRAS and NRAS mutations has entered routine clinical practice. The role of the other markers still needs to be validated. The ongoing benefit of anti-EGFR therapy could be indicated by specific clinical parameters measured after the initiation of targeted therapy, including early tumor shrinkage, the deepness of the response, or hypomagnesemia. The accuracy of predictive dia­gnostic tools could be also increased by examining a combination of predictive markers using next generation sequencing methods. However, unjustified investigation of many molecular markers should be resisted as this may complicate interpretation of the results, particularly in terms of their specific clinical relevance. Aim: The aim of this review is to describe current possibilities with respect to predicting responses to EGFR blockade in the context of the EGFR pathway, and the utilization of such results in routine clinical practice. Key words: colorectal cancer – cetuximab – panitumumab – EGFR – KRAS – BRAF The authors declare they have no potential conflicts of interest concerning drugs, products, or services used in the study. The Editorial Board declares that the manuscript met the ICMJE recommendation for biomedical papers. Submitted: 20. 3. 2016 Accepted: 19. 4. 2016

• MeSH
• analýza přežití MeSH
• erbB receptory * antagonisté a inhibitory   MeSH
• exantém MeSH
• exprese genu MeSH
• fosfatidylinositol-3-kinasy genetika   MeSH
• fosfohydroláza PTEN genetika   MeSH
• genetické testování * MeSH
• kolorektální nádory * genetika   terapie   MeSH
• kombinovaná terapie MeSH
• lidé MeSH
• metastázy nádorů genetika   MeSH
• nádorové biomarkery MeSH
• nedostatek hořčíku MeSH
• prediktivní hodnota testů MeSH
• prognóza MeSH
• protokoly protinádorové kombinované chemoterapie MeSH
• protoonkogenní proteiny B-raf genetika   MeSH
• ras proteiny * genetika   MeSH
• retrospektivní studie MeSH
• výsledek terapie MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Cellular senescence leads to decreased tissue regeneration and inflammation and is associated with diabetes, neurodegenerative diseases, and tumorigenesis. However, the mechanisms of cellular senescence are not fully understood. Emerging evidence has indicated that c-Jun N-terminal kinase (JNK) signaling is involved in the regulation of cellular senescence. JNK can downregulate hypoxia inducible factor-1α to accelerate hypoxia-induced neuronal cell senescence. The activation of JNK inhibits mTOR activity and triggers autophagy, which promotes cellular senescence. JNK can upregulate the expression of p53 and Bcl-2 and accelerates cancer cell senescence; however, this signaling also mediates the expression of amphiregulin and PD-LI to achieve cancer cell immune evasion and prevents their senescence. The activation of JNK further triggers forkhead box O expression and its target gene Jafrac1 to extend the lifespan of Drosophila. JNK can also upregulate the expression of DNA repair protein poly ADP-ribose polymerase 1 and heat shock protein to delay cellular senescence. This review discusses recent advances in understanding the function of JNK signaling in cellular senescence and includes a comprehensive analysis of the molecular mechanisms underlying JNK-mediated senescence evasion and oncogene-induced cellular senescence. We also summarize the research progress in anti-aging agents that target JNK signaling. This study will contribute to a better understanding of the molecular targets of cellular senescence and provides insights into anti-aging, which may be used to develop drugs for the treatment of aging-related diseases.

• MeSH
• hypoxie MeSH
• JNK mitogenem aktivované proteinkinasy * metabolismus   MeSH
• lidé MeSH
• MAP kinasový signální systém MeSH
• signální transdukce * MeSH
• stárnutí buněk MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• přehledy MeSH