Východiska: Metastazování, relaps a rezistence k chemoterapii jsou celosvětově hlavními důvody většiny úmrtí na malignitu. Proces metastazování je velmi precizně organizovaný a orgánově specifický. Mechanizmy metastazování nejsou stále přesně známy, ale některé z nich jsou zprostředkovány programem epitelo-mezenchymální tranzice (EMT). Poprvé byl tento proces popsán v průběhu embryogeneze. Tento buněčný program vede ke změně epiteliálního fenotypu buňky v mezenchymální typ. V průběhu této tranzice dochází ke změně tvaru epiteliální buňky, která ztrácí svoji polaritu, závislost na buněčných spojích a celou řadu epiteliálních markerů. Naopak získává vřetenovitý tvar a stává se mechanicky odolnější a je schopnější vycestovat z buněčné kolonie. Tato změna fenotypu je provázena i změnou exprese proteinů a genů, jako jsou transkripční faktory, cadheriny, cateniny, matrix metaloproteinázy nebo receptory pro růstové faktory. Známky aktivace programu EMT byly již popsány v různých typech nádorů, např. u karcinomu prsu, vaječníků, karcinomu tlustého střeva a jícnu, a jejich přítomnost byla asociována s horší prognózou onemocnění a častějším metastatickým šířením. Také rezistence k cytostatické léčbě je spojována s reaktivací těchto embryonálních procesů. Porozumění tomuto programu je nezbytné k lepšímu chápání procesu progrese maligního onemocnění, stejně jako k hledání nových léčebných cílů a prognostických faktorů. Závěr: Tento článek shrnuje dosud známé molekulární kaskády zapojené do procesu EMT u nádorových onemocnění.

Background: Metastasis, recurrence, and resistance to chemotherapy are leading causes of the majority of cancer-related mortality worldwide. The process of metastasis can be artificially divided into a series of sequential, highly organized, and organ-specific steps. The underlying mechanisms are still poorly understood, but are believed to be mediated by epithelial-mesenchymal transition (EMT). First described in embryogenesis, EMT is a cellular reprogramming process in which epithelial cells acquire a mesenchymal phenotype. During this transformation, epithelial cells lose their shape, epithelial markers, and ability to grow in colonies. They acquire a spindle-shaped morphology and exhibit more motile and invasive behavior. These phenotypic changes are associated with modifications in different interconnected protein and gene families, such as transcription factors, cadherins, catenins, matrix metalloproteases, and growth receptors. EMT has been observed in many cancers, such as breast, ovarian, colon, and esophageal cancers, and is associated with poor prognosis and metastasis. Also, resistance to cytotoxic treatments is associated with reactivation of embryonic programs. Understanding this process is necessary to provide a better understanding of cancer progression and could lead to the development of new therapeutic or prognostic strategies for the treatment of cancer. Conclusion: This article summarizes the known molecular pathways involved in EMT in cancer.

• MeSH
• epitelo-mezenchymální tranzice * fyziologie   účinky léků   MeSH
• fenotyp MeSH
• hojení ran MeSH
• kadheriny fyziologie   MeSH
• karcinom patologie   MeSH
• lékové interakce MeSH
• lidé MeSH
• metastázy nádorů * patofyziologie   MeSH
• mikro RNA fyziologie   MeSH
• nádorová transformace buněk MeSH
• neurální trubice fyziologie   MeSH
• proliferace buněk fyziologie   MeSH
• receptory Notch MeSH
• signální dráha Wnt fyziologie   MeSH
• transkripční faktory fyziologie   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• přehledy MeSH

Cíl studie: V reprodukčním období ženy dochází k opakovaným a častým strukturálním a funkčním proměnám endometria. Schopnost regenerace, remodelace a diferenciace je předpokladem receptivity endometria, implantace a vývoje embrya. Důležitým faktorem těchto procesů je vzájemná přeměna mezenchymálního a epiteliálního fenotypu endometriálních buněk – epiteliálně- -mezenchymální transice (EMT = epitelial-mesenchymal transition) a mezenchymálně-epiteliální transice (MET = mesenchymal-epithelial transition). Cílem práce je prezentovat současné poznatky o vzájemné přeměně epiteliálních a mezenchymální buněk děložní sliznice a jejich možném vlivu na poruchy plodnosti. Typ studie: Přehledová práce. Název a sídlo pracoviště: Gynekologicko-porodnická klinika Lékařské fakulty Masarykovy univerzity a FN Brno; Porodnicko-gynekologická klinika FN a LF UP Olomouc. Metodika: Literární rešerše databáze PubMed publikované do února 2019 s termíny zaměřenými na „endometrial receptivity“, „embryo implantation“, „endometrial regeneration“, „mesenchymal–epithelial transition/transformation“. Výsledky: Bylo prokázáno, že stromální buňky se podílejí na regeneraci nejen stromatu, ale také epitelu endometria. V průběhu decidualizace působením ovariálních steroidů a dalších faktorů probíhá MET, fibroblasty stromatu získávají postupně vlastnosti epiteliálních buněk – morfologicky i funkčně (sekreční endoplazmatické retikulum, pevné intercelulární spoje). V průběhu implantace embrya vlivem interakce trofoblastu s decidualizovaným endometriem dochází k přeměně epiteliálních buněk na mezenchymální (EMT), které jsou schopny migrace a regulace pronikajícího trofoblastu. Závěr: Vzájemná přeměna stromálních a epiteliálních buněk endometria je nezbytná pro fyziologickou funkci děložní sliznice včetně implantace a vývoje embrya.

Objective: During reproductive age of a woman, endometrium undergoes frequent stuctural and functional changes. Abilities of regeneration, remodelation and differentiation are precondition of endometrial receptivity and implantation and development of an embryo. These processes are conditioned by mutual transformation between mesenchymal and epithelial fenotype of endometrial cells: epithelial-mesenchymal transition (EMT) and mesenchymal-epithelial transition (MET). The aim of this study is to present contemporary knowledge of transformation between epithelial and mesenchymal endometrial cells and its influence on human fertility. Design: Review article. Setting: Department of Obstetrics and Gynecology, Faculty of Medicine, Masaryk university and University Hospital Brno; Department of Obstetrics and Gynecology, University Hospital Faculty of Medicine, Palacky University, Olomouc. Methods: PubMed was searched for articles in English indexed until February 2019 with terms of „endometrial receptivity“, „embryo implantation“, „endometrial regeneration“, „mesenchymal-epithelial transition/ transformation“. Results: It has been proved, that mesenchymal stromal cells participate on regeneration of not only the endometrial stroma, but also of the epithelium. During endometrial decidualisation under influence of ovarian steroids, the MET is under way. Stromal fibroblasts gain the morfological and functional properties of epithelial cells. During implantaion of an embryo, the trofoblast interacts with decidualised endometrium. Epithelial cells transform into mesenchymal (EMT), which mediate the growth of trofoblast. Conclusion: Mutual transformation between stromal and epithelial cells in essential for normal function of endometrium and implantation and development of an embryo.

• Klíčová slova
• mezenchymálněepiteliální transformace buněk, receptivita endometria, regenerace endometria,
• MeSH
• endometrium * fyziologie   MeSH
• implantace embrya MeSH
• lidé MeSH
• Check Tag
• lidé MeSH
• ženské pohlaví MeSH
• Publikační typ
• práce podpořená grantem MeSH
• přehledy MeSH

Cystic fibrosis (CF) is a monogenetic disease resulting from mutations in the Cystic Fibrosis Transmembrane conductance Regulator (CFTR) gene encoding an anion channel. Recent evidence indicates that CFTR plays a role in other cellular processes, namely in development, cellular differentiation and wound healing. Accordingly, CFTR has been proposed to function as a tumour suppressor in a wide range of cancers. Along these lines, CF was recently suggested to be associated with epithelial-mesenchymal transition (EMT), a latent developmental process, which can be re-activated in fibrosis and cancer. However, it is unknown whether EMT is indeed active in CF and if EMT is triggered by dysfunctional CFTR itself or a consequence of secondary complications of CF. In this study, we investigated the occurrence of EMT in airways native tissue, primary cells and cell lines expressing mutant CFTR through the expression of epithelial and mesenchymal markers as well as EMT-associated transcription factors. Transepithelial electrical resistance, proliferation and regeneration rates, and cell resistance to TGF-β1induced EMT were also measured. CF tissues/cells expressing mutant CFTR displayed several signs of active EMT, namely: destructured epithelial proteins, defective cell junctions, increased levels of mesenchymal markers and EMT-associated transcription factors, hyper-proliferation and impaired wound healing. Importantly, we found evidence that the mutant CFTR triggered EMT was mediated by EMT-associated transcription factor TWIST1. Further, our data show that CF cells are over-sensitive to EMT but the CF EMT phenotype can be reversed by CFTR modulator drugs. Altogether, these results identify for the first time that EMT is intrinsically triggered by the absence of functional CFTR through a TWIST1 dependent mechanism and indicate that CFTR plays a direct role in EMT protection. This mechanistic link is a plausible explanation for the high incidence of fibrosis and cancer in CF, as well as for the role of CFTR as tumour suppressor protein.

• MeSH
• cystická fibróza metabolismus   patologie   MeSH
• epitelo-mezenchymální tranzice MeSH
• HEK293 buňky MeSH
• jaderné proteiny metabolismus   MeSH
• kultivované buňky MeSH
• lidé MeSH
• onkogeny genetika   MeSH
• protein CFTR metabolismus   MeSH
• transkripční faktor Twist metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

Preterm, prelabor rupture of the human fetal membranes (pPROM) is involved in 40% of spontaneous preterm births worldwide. Cellular-level disturbances and inflammation are effectors of membrane degradation, weakening, and rupture. Maternal risk factors induce oxidative stress (OS), senescence, and senescence-associated inflammation of the fetal membranes as reported mechanisms related to pPROM. Inflammation can also arise in fetal membrane cells (amnion/chorion) due to OS-induced autophagy and epithelial-mesenchymal transition (EMT). Autophagy, EMT, and their correlation in pPROM, along with OS-induced autophagy-related changes in amnion and chorion cells in vitro, were investigated. Immunocytochemistry staining of cytokeratin-18 (epithelial marker)/vimentin (mesenchymal marker) and proautophagy-inducing factor LC3B were performed in fetal membranes from pPROM, term not in labor, and term labor. Ultrastructural changes associated with autophagy were verified by transmission electron microscopy of the fetal membranes and in cells exposed to cigarette smoke extract (an OS inducer). EMT and LC3B staining was compared in the chorion from pPROM versus term not in labor. Transmission electron microscopy confirmed autophagosome formation in pPROM amnion and chorion. In cell culture, autophagosomes were formed in the amnion with OS treatment, while autophagosomes were accumulated in both cell types with autophagy inhibition. This study documents the association between pPROMs and amniochorion autophagy and EMT, and supports a role for OS in inducing dysfunctional cells that increase inflammation, predisposing membranes to rupture.

• MeSH
• autofagie MeSH
• epitelo-mezenchymální tranzice MeSH
• extraembryonální obaly * metabolismus   MeSH
• lidé MeSH
• novorozenec MeSH
• předčasný odtok plodové vody * metabolismus   MeSH
• zánět patologie   MeSH
• Check Tag
• lidé MeSH
• novorozenec MeSH
• ženské pohlaví MeSH
• Publikační typ
• časopisecké články MeSH

The cell surface glycoprotein Trop-2 is commonly overexpressed in carcinomas and represents an exceptional antigen for targeted therapy. Here, we provide evidence that surface Trop-2 expression is functionally connected with an epithelial phenotype in breast and prostate cell lines and in patient tumor samples. We further show that Trop-2 expression is suppressed epigenetically or through the action of epithelial-to-mesenchymal transition transcription factors and that deregulation of Trop-2 expression is linked with cancer progression and poor patient prognosis. Moreover, our data suggest that the cancer plasticity-driven intratumoral heterogeneity in Trop-2 expression may significantly contribute to response and resistance to therapies targeting Trop-2-expressing cells.

• MeSH
• antigeny nádorové genetika   metabolismus   MeSH
• CD antigeny biosyntéza   MeSH
• epitelo-mezenchymální tranzice fyziologie   MeSH
• epitelové buňky metabolismus   MeSH
• kadheriny biosyntéza   MeSH
• karcinom patologie   MeSH
• lidé MeSH
• metylace DNA genetika   MeSH
• molekuly buněčné adheze genetika   metabolismus   MeSH
• myši inbrední BALB C MeSH
• myši MeSH
• nádorové buněčné linie MeSH
• nádory prostaty mortalita   patologie   MeSH
• nádory prsu mortalita   patologie   MeSH
• progrese nemoci MeSH
• xenogenní modely - testy protinádorové aktivity MeSH
• zvířata MeSH
• Check Tag
• lidé MeSH
• mužské pohlaví MeSH
• myši MeSH
• ženské pohlaví MeSH
• zvířata MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

SIGNIFICANCE: Machine learning is increasingly being applied to the classification of microscopic data. In order to detect some complex and dynamic cellular processes, time-resolved live-cell imaging might be necessary. Incorporating the temporal information into the classification process may allow for a better and more specific classification. AIM: We propose a methodology for cell classification based on the time-lapse quantitative phase images (QPIs) gained by digital holographic microscopy (DHM) with the goal of increasing performance of classification of dynamic cellular processes. APPROACH: The methodology was demonstrated by studying epithelial-mesenchymal transition (EMT) which entails major and distinct time-dependent morphological changes. The time-lapse QPIs of EMT were obtained over a 48-h period and specific novel features representing the dynamic cell behavior were extracted. The two distinct end-state phenotypes were classified by several supervised machine learning algorithms and the results were compared with the classification performed on single-time-point images. RESULTS: In comparison to the single-time-point approach, our data suggest the incorporation of temporal information into the classification of cell phenotypes during EMT improves performance by nearly 9% in terms of accuracy, and further indicate the potential of DHM to monitor cellular morphological changes. CONCLUSIONS: Proposed approach based on the time-lapse images gained by DHM could improve the monitoring of live cell behavior in an automated fashion and could be further developed into a tool for high-throughput automated analysis of unique cell behavior.

• MeSH
• algoritmy MeSH
• časosběrné zobrazování MeSH
• epitelo-mezenchymální tranzice * MeSH
• holografie * MeSH
• strojové učení MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

TGFβ has roles in inflammation, wound healing, epithelial to mesenchymal transition (EMT), and cancer stem cell states, and acts as a tumor suppressor gene for squamous cell carcinoma (SCC). SCCs are also characterized by high levels of ΔNp63, which induces epithelial cell phenotypes and maintains squamous stem cells. Previous studies indicate a complex interplay between ΔNp63 and TGFβ signaling, with contradictory effects reported. We investigated the effects of TGFβ on p63 isoform proteins and mRNAs in non-malignant squamous and SCC cells, and the role of either canonical or non-canonical TGFβ signaling pathways. TGFβ selectively increased ΔNp63 protein levels in non-malignant keratinocytes in association with SMAD3 activation and was prevented by TGFβ receptor inhibition, indicating activation of canonical TGFβ pathway signaling. TP63 isoform mRNAs showed discordance from protein levels, with an initial increase in both TAP63 and ΔNP63 mRNAs followed by a decrease at later times. These data demonstrate complex and heterogeneous effects of TGFβ in squamous cells that depend on the extent of canonical TGFβ pathway aberrations. The interplay between TGFβ and p63 is likely to influence the magnitude of EMT states in SCC, with clinical implications for tumor progression and response to therapy.

• MeSH
• epitelo-mezenchymální tranzice * genetika   MeSH
• epitelové buňky metabolismus   MeSH
• lidé MeSH
• protein - isoformy genetika   metabolismus   MeSH
• spinocelulární karcinom * genetika   metabolismus   MeSH
• transformující růstový faktor beta MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Lower respiratory tract infection due to Pseudomonas aeruginosa has become increasingly challenging, resulting in a worse morbidity and mortality. Airway remodeling is a common phenomenon in this process, to which epithelial-mesenchymal transition (EMT) may contribute as an important promoter. Previous studies showed that epithelium-specific integrin αvβ6-mediated EMT was involved in pulmonary fibrosis via transforming growth factor-β1 (TGF-β1) signaling, but whether integrin αvβ6 plays a role in the P. aeruginosa-associated airway remodeling remains unknown. BEAS-2B cells were incubated with lipopolysaccharide (LPS) from P. aeruginosa in the presence or the absence of integrin αvβ6-blocking antibodies. Morphologic changes were observed by an inverted microscopy. The EMT markers were detected using Western blotting and immunofluorescence. The activation of TGF-β1-Smad2/3 signaling pathway was assessed. Furthermore, matrix metalloproteinase (MMP)-2 and -9 in the medium were measured using ELISA. P. aeruginosa's LPS decreased the expression of the epithelial marker E-cadherin and promoted the mesenchymal markers, vimentin and α-smooth muscle actin in BEAS-2B cells. The expression of integrin αvβ6 was significantly increased during EMT process. Blocking integrin αvβ6 could attenuate P. aeruginosa's LPS-induced EMT markers' expression via TGF-β1-Smad2/3 signaling pathway. Furthermore, blocking integrin αvβ6 could prevent morphologic changes and oversecretion of MMP-2 and -9. Integrin αvβ6 mediates epithelial-mesenchymal transition in human bronchial epithelial cells induced by lipopolysaccharides of P. aeruginosa via TGF-β1-Smad2/3 signaling pathway and might be a promising therapeutic target for P. aeruginosa-associated airway remodeling.

• MeSH
• antigeny nádorové genetika   metabolismus   MeSH
• epitelo-mezenchymální tranzice * MeSH
• epitelové buňky cytologie   účinky léků   metabolismus   MeSH
• integriny genetika   metabolismus   MeSH
• lidé MeSH
• lipopolysacharidy metabolismus   MeSH
• matrixové metaloproteinasy genetika   metabolismus   MeSH
• protein Smad2 genetika   metabolismus   MeSH
• protein Smad3 genetika   metabolismus   MeSH
• pseudomonádové infekce genetika   metabolismus   mikrobiologie   patofyziologie   MeSH
• Pseudomonas aeruginosa metabolismus   MeSH
• signální transdukce MeSH
• transformující růstový faktor beta1 genetika   metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH

Hematopoietic stem cells derived from pluripotent stem cells could be used as an alternative to bone marrow transplants. Deriving these has been a long-term goal for researchers. However, the success of these efforts has been limited with the cells produced able to engraft in the bone marrow of recipient animals only in very low numbers. There is evidence that defects in the migratory and homing capacity of the cells are due to mis-regulation of miRNA expression and are responsible for their failure to engraft. We compared the miRNA expression profile of hematopoietic progenitors derived from pluripotent stem cells to those derived from bone marrow and found that numerous miRNAs are too highly expressed in hematopoietic progenitors derived from pluripotent stem cells, and that most of these are inhibitors of epithelial-mesenchymal transition or metastasis (including miR-200b, miR-200c, miR-205, miR-148a, and miR-424). We hypothesize that the high expression of these factors, which promote an adherent phenotype, may be causing the defect in hematopoietic differentiation. However, inhibiting these miRNAs, individually or in multiplex, was insufficient to improve hematopoietic differentiation in vitro, suggesting that other miRNAs and/or genes may be involved in this process. Stem Cells 2018;36:55-64.

• MeSH
• buněčná diferenciace MeSH
• down regulace MeSH
• epitelo-mezenchymální tranzice genetika   MeSH
• hematopoetické kmenové buňky metabolismus   MeSH
• lidé MeSH
• mikro RNA genetika   MeSH
• pluripotentní kmenové buňky metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH
• práce podpořená grantem MeSH

BACKGROUND: During cancer progression, epithelial cancer cells can be reprogrammed into mesenchymal-like cells with increased migratory potential through the process of epithelial-mesenchymal transition (EMT), representing an essential step of tumor progression towards metastatic state. AGR2 protein was shown to regulate several cancer-associated processes including cellular proliferation, survival and drug resistance. METHODS: The expression of AGR2 was analyzed in cancer cell lines exposed to TGF-β alone or to combined treatment with TGF-β and the Erk1/2 inhibitor PD98059 or the TGF-β receptor specific inhibitor SB431542. The impact of AGR2 silencing by specific siRNAs or CRISPR/Cas9 technology on EMT was investigated by western blot analysis, quantitative PCR, immunofluorescence analysis, real-time invasion assay and adhesion assay. RESULTS: Induction of EMT was associated with decreased AGR2 along with changes in cellular morphology, actin reorganization, inhibition of E-cadherin and induction of the mesenchymal markers vimentin and N-cadherin in various cancer cell lines. Conversely, induction of AGR2 caused reversion of the mesenchymal phenotype back to the epithelial phenotype and re-acquisition of epithelial markers. Activated Smad and Erk signaling cascades were identified as mutually complementary pathways responsible for TGF-β-mediated inhibition of AGR2. CONCLUSION: Taken together our results highlight a crucial role for AGR2 in maintaining the epithelial phenotype by preventing the activation of key factors involved in the process of EMT.

• MeSH
• buněčná adheze genetika   MeSH
• epitelo-mezenchymální tranzice účinky léků   genetika   MeSH
• genový knockdown MeSH
• kadheriny metabolismus   MeSH
• lidé MeSH
• nádorové buněčné linie MeSH
• pohyb buněk genetika   MeSH
• proteiny Smad metabolismus   MeSH
• proteiny genetika   MeSH
• regulace genové exprese u nádorů účinky léků   MeSH
• signální transdukce účinky léků   MeSH
• transformující růstový faktor beta farmakologie   MeSH
• vimentin metabolismus   MeSH
• Check Tag
• lidé MeSH
• Publikační typ
• časopisecké články MeSH