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PubMed2006

Relevance of the stroma and epithelial-mesenchymal transition (EMT) for the rheumatic diseases.

Epithelial-mesenchymal transition (EMT) is a term applied to the process whereby cells undergo a switch from an epithelial phenotype with tight junctions, lateral, apical, and basal membranes, and lack of mobility into mesenchymal cells that have loose interactions with other cells, are non-polarized, motile and produce an extracellular matrix. The importance of this process was initially recognized from a very early step in embryology, but more recently as a potential mechanism for the progression and spread of epithelial cancers. As the sequence of morphological changes has become understood in molecular terms, diseases characterized by alterations in stromal elements and fibrosis are being considered as examples of EMT. This review will focus on the pathogenetic features of immune-mediated renal disease, systemic sclerosis and rheumatoid arthritis that could be explained by EMT.

Arthritis, Rheumatoid↗
PubMed2026

ENTPD3 as a novel regulator of endometrial receptivity: suppressing EMT via the ATP-P2Y2 axis in patients with recurrent implantation failure.

BACKGROUND: Recurrent implantation failure (RIF) remains a major challenge in assisted reproductive technology and is primarily attributed to impaired endometrial receptivity. Despite its clinical significance, the precise mechanisms underlying RIF remain inadequately understood. METHODS: Single-cell RNA sequencing (scRNA-seq) was performed on endometrial samples from patients with RIF and healthy controls during the secretory phase using the 10X Genomics Chromium platform. The expression and localization of ectonucleoside triphosphate diphosphohydrolase 3 (ENTPD3) in the window of implantation (WOI) in the endometrium were examined using real-time quantitative polymerase chain reaction (RT-qPCR), western blotting, and immunohistochemistry (IHC). A mouse model with ENTPD3 overexpression was utilized to assess embryo implantation in vivo, and an in vitro blastocyst adhesion assay was performed to evaluate endometrial receptivity. Additionally, Ishikawa cells were transduced with an ENTPD3 recombinant adenovirus to explore the underlying molecular mechanisms. RESULTS: ENTPD3 expression was significantly upregulated in the endometria of patients with RIF during the WOI, and its apical surface localization in endometrial epithelial cells was confirmed by single-cell data and IHC. Functional studies demonstrated that ENTPD3 overexpression impaired endometrial receptivity by suppressing epithelial-mesenchymal transition (EMT). In vivo, ENTPD3 overexpression markedly reduced endometrial receptivity and inhibited embryo implantation in mice. Consistently, in vitro assays revealed that ENTPD3 overexpression diminished blastocyst adhesion to endometrial epithelial cells. Mechanistically, ENTPD3 hydrolyzes ATP, thereby suppressing EMT via the P2Y2 signaling pathway and ultimately disrupting endometrial receptivity. CONCLUSIONS: Dysregulated ENTPD3 expression contributes to RIF pathogenesis by impairing endometrial receptivity through ATP hydrolysis-mediated suppression of EMT via P2Y2 signaling. These findings highlight ENTPD3 as a potential therapeutic target for improving implantation success in affected patients.

Female↗
PubMed1983

Examining EMT education: a shift in emphasis.

This article examines several aspects of EMT education in the United States. It deemphasizes the instructor's total dependence on objectives and examination results and encourages EMT educators to begin looking at the actual instructional process itself. This concept is encouraged through attainment by individual training programs of AMA accreditation (EMT-Paramedic level only), development of a spirit of collegiality and of inquiry about the practice of teaching, and utilization of formalized primary training and continuing education courses on improving instruction technique.

Allied Health Personnel↗
PubMed2003

New approach to generalized two-dimensional correlation spectroscopy. II: Eigenvalue manipulation transformation (EMT) for noise suppression.

This paper introduces the concept of eigenvalue manipulating transformation (EMT) of a data matrix for noise suppression in two-dimensional (2D) correlation spectroscopy. The FT-IR spectra of a polystyrene/methyl ethyl ketone/toluene solution mixture during the solvent evaporation process, to which were added a substantial amount of artificial noise, have been analyzed. By uniformly raising the power of a set of eigenvalues, the major eigenvalues become more prominent. As a consequence, minor eigenvectors representing the noise component are no longer strongly represented in the reconstructed data. This EMT operation is similar to the simple truncation of noise-dominated minor factors practiced in standard principal component analysis (PCA), as demonstrated in our preceding paper on PCA-2D correlation spectroscopy. The effect of this new EMT scheme is more gradual, with attractive flexibility to continuously fine-tune the balance between the desired noise reduction effect and the retention of pertinent spectral information.

Algorithms↗
PubMed1992

Killing of EMT-6 cells by decays from isotopes incorporated on sensitizer adducts.

EMT-6 tumor cell killing by decays from 3H and 125I incorporated by adduct formation of radiolabeled sensitizers was studied in vitro. Hypoxic radiosensitizers become covalently bound to cellular molecules after metabolic reduction, and EMT-6 tumor cells can tolerate over 10(9) adducts/cell of misonidazole without loss of colony-forming ability. Cells were incubated under hypoxic conditions in the presence of [3H]misonidazole or [125I]iodoazomycinriboside for various times and the amounts of bound 3H and 125I were determined. Cells were stored as monolayers at 22 degrees C, in suspension culture at 4 degrees C, and frozen in complete medium plus 8% DMSO at -196 degrees C for various times to facilitate the accumulation of radioactive decays before plating in vitro for colony-forming assays at 37 degrees C. At 22 degrees C in monolayer culture, EMT-6 tumor cells tolerated 950 and 1720 decays/cell of 3H and 125I, respectively, without evidence of radiotoxicity. This number of decays/cell over the exposure times used represents 1.54 x 10(6) 3H/cell and 8.4 x 10(4) 125I/cell, respectively. Significant cell killing was detected after similar amounts of isotope decay when cells were held at 4 degrees C. When cells were frozen in the presence of 8% DMSO, they were more resistant to inactivation by isotope decays or by gamma rays than cells in liquid phase at 4 degrees C. These data suggest that selective hypoxic tumor cell suicide by 3H or 125I decays from bound sensitizer at 37 degrees C will be an inefficient process, at least for drugs with specific activities as tested. These data are consistent with data on cell inactivation by isotopes incorporated into cells by other procedures.

Animals↗
PubMed1979

A transferrin-mediated uptake of gallium-67 by EMT-6 sarcoma. I. Studies in tissue culture.

We have studied the in vitro uptake of gallium-67 by exponentially growing EMT-6 sarcoma cells in long-term tissue culture. In this system, the addition of transferrin to the medium was required before an appreciable cellular uptake of Ga-67 occurred. The transferrin effect was complex, with an initial stimulation to a peak cell-to-medium ratio of 8--10:1 at low concentrations of transferrin (0.2 mg/ml), followed by a gradual decline in uptake as transferrin in the medium was increased further. EMT-6 tumor-cell uptake of Ga-67 was probably mediated by a specific cellular receptor for transferrin. Scatchard analysis of the EMT-6 cellular binding of human transferrin labeled with iodine-125 indicated a cellular receptor with affinity for transferrin of 5 X 10(6) l/mole and abundance of 500,000 receptors per cell. Over the experimental range of transferrin concentration in the medium, the observed uptake of Ga-67 was closely correlated with the degree of formation of Ga-67-labeled transferrin and the fraction of transferrin bound to the cellular receptor (N = 69, r = 0.86, p less than 0.0001).

Animals↗
PubMed2025

miR-9-5p/HMMR regulates the tumorigenesis and progression of clear cell renal cell carcinoma through EMT and JAK1/STAT1 signaling pathway.

BACKGROUND: The most common malignant type of kidney cancer is clear cell renal cell carcinoma (ccRCC). The expression levels of hyaluronan-mediated motility receptor (HMMR) in many tumor types are significantly elevated. HMMR is closely associated with tumor-related progression, treatment resistance, and poor prognosis, and has yet to be fully investigated in terms of its expression patterns and molecular mechanisms of action in ccRCC. Further research is imperative to elucidate these aspects. METHODS: We used The Cancer Genome Atlas (TCGA) database to preliminarily investigate HMMR expression and function in ccRCC and the data for 19 samples from the NCBI GEO database (GSE207493) for single-cell analysis. We assessed the differential expression level of HMMR between ccRCC cancerous tissues and their matched non-tumor tissues. Subsequently, a series of in vivo and in vitro experiments were designed to elucidate the biological function of HMMR in ccRCC, including Transwell assays, CCK-8 assays, clone formation assays and subcutaneous xenograft experiments in nude mice. Through bioinformatics analysis, we identified potential microRNAs (miRNAs) that may regulate HMMR, as well as the possible signaling pathways involved. Finally, we conducted a series of cellular functional experiments to validate our hypotheses regarding the HMMR axis. RESULTS: HMMR expression was significantly up-regulated in tumor tissues of ccRCC patients, and elevated HMMR expression level showed a strong correlation with ccRCC progression and adverse prognoses of patients. Knocking down HMMR inhibited the proliferative and migratory abilities of ccRCC cells, while its overexpression amplified these oncogenic properties. In nude mice model, reduced HMMR expression inhibited ccRCC tumor proliferation in vivo. Furthermore, overexpression of an upstream transcriptional regulator, miR-9-5p, effectively downregulated HMMR expression and thus impeded ccRCC cells proliferation and migration. HMMR might influence ccRCC growth via the Epithelial-Mesenchymal Transition (EMT) pathway and the Janus Kinase 1/Signal Transducer and Activator of Transcription 1 (JAK1/STAT1) pathway. CONCLUSIONS: HMMR is overexpressed in ccRCC, and there is a significant link between high HMMR expression and tumor progression, as well as poor patient prognosis. Specifically, HMMR could be targeted and inhibited by miR-9-5p and might modulate the tumorigenesis and progression of ccRCC through both EMT and JAK1/STAT1 signaling pathway.

Carcinoma, Renal Cell↗
PubMed2006

Tamoxifen resistance in MCF7 cells promotes EMT-like behaviour and involves modulation of beta-catenin phosphorylation.

We have previously demonstrated that, following acquisition of endocrine resistance, breast cancer cells display an altered growth rate together with increased aggressive behaviour in vitro. Since dysfunctional cell-cell adhesive interactions can promote an aggressive phenotype, we investigated the integrity of this protein complex in our breast cancer model of tamoxifen resistance. In culture, tamoxifen-resistant MCF7 (TamR) cells grew as loosely packed colonies with loss of cell-cell junctions and demonstrated altered morphology characteristic of cells undergoing epithelial-to-mesenchymal transition (EMT). Neutralising E-cadherin function promoted the invasion and inhibited the aggregation of endocrine-sensitive MCF7 cells, whilst having little effect on the behaviour of TamR cells. Additionally, TamR cells had increased levels of tyrosine-phosphorylated beta-catenin, whilst serine/threonine-phosphorylated beta-catenin was decreased. These cells also displayed loss of association between beta-catenin and E-cadherin, increased cytoplasmic and nuclear beta-catenin and elevated transcription of beta-catenin target genes known to be involved in tumour progression and EMT. Inhibition of EGFR kinase activity in TamR cells reduced beta-catenin tyrosine phosphorylation, increased beta-catenin-E-cadherin association and promoted cell-cell adhesion. In such treated cells, the association of beta-catenin with Lef-1 and the transcription of c-myc, cyclin-D1, CD44 and COX-2 were also reduced. These results suggest that homotypic adhesion in tamoxifen-resistant breast cancer cells is dysfunctional due to EGFR-driven modulation of the phosphorylation status of beta-catenin and may contribute to an enhanced aggressive phenotype and transition towards a mesenchymal phenotype in vitro.

Antineoplastic Agents, Hormonal↗
PubMed1980

Linkage of genetic markers emt and chr in Chinese hamster cells.

Segregation analysis in somatic cell hybrids has been used to demonstrate linkage in the CHO cell line between two autosomal loci, emt (defined by mutants resistant to the protein synthesis inhibitor emetine) and chr (defined by mutants resistant to chromate). In both cases drug resistance behaves recessively in hybrids and linkage has been determined by examining the recovery of the mutant phenotypes from hybrids of the type CHO emtr chrr x CHO emt+ chr+. Of 24 independent segregants selected for resistance to emetine, all 24 were also resistant to chromate while only 18 out of 24 segregants selected for chromate resistance were also resistant to emetine. The biological significane of this "polarity effect" is currently unknown but may be related to the molecular mechanism or mechanisms by which segregation occurs in hybrid cells. In this and the accompanying paper (Worton, Duff, and Campbell) we discuss a number of possible mechanisms of segregation, of which we believe mitotic recombination to be most consistent with the available data.

Animals↗
PubMed1993

Establishment and results of an EMT-D program in a two-tiered physician-escorted rescue system. The experience in Berlin, Germany.

Early defibrillation by emergency medical technicians or even less qualified personnel has been shown to improve survival rates for out-of-hospital cardiac arrest caused by ventricular fibrillation. It has been questioned whether these favourable results can be applied within the context of physician-attended emergency medical systems. Taking into consideration the results of a pilot study and after a careful analysis of the logistic and epidemiological background, the first German EMT-D program was introduced in the former West Berlin in December 1988. The first 2 years of experience with 499 technician-initiated resuscitation attempts in which the mobile intensive care unit of Klinikum Steglitz was involved, confirmed the results of the pilot study with an improved long-term survival rate (18%) for patients with ventricular fibrillation. We conclude that EMT defibrillation should be introduced in emergency physician-attended two-tiered emergency medical systems, whenever a thorough analysis of the existing rescue systems exhibits a 'relevant frequency' of resuscitation and response interval of 15 min or less.

Ambulances↗
PubMed1994

Observation of spinning sidebands in the 27Al magic-angle spinning nuclear magnetic resonance spectra of FAU and EMT structure-type zeolites.

Wide-band (1 MHz) 27Al magic-angle spinning nuclear magnetic resonance (MAS NMR) spectra were recorded for zeolite Y (Si/Al approximately 2.7) and high-silica (Si/Al approximately 3.8) FAU, EMT and mixed FAU-EMT structure-type zeolites synthesized using 15-crown-5 and/or 18-crown-6 ethers as templating agents. Spinning sidebands (related to first-order quadrupolar effects affecting satellite transitions) are observed in the spectra of the four calcined and fully rehydrated samples, reflecting a high symmetry of the framework aluminium atom surrounded by four oxygens. It is shown that the intensity of the spinning sidebands progressively increases after calcination and rehydration of the samples, indicating that the restoration of the high local order around the 27Al nuclei is rather slow. On the other hand, second-order quadrupolar interactions rapidly decrease upon rehydration of the calcined samples which is achieved within one day, as indicated by thermogravimetry. Some hypotheses are proposed to explain such a difference, and the role of water is also discussed.

Aluminum↗
PubMed2006

Total syntheses of three copper (II) tetracarboranylphenylporphyrins containing 40 or 80 boron atoms and their biological properties in EMT-6 tumor-bearing mice.

Three carboranyltetraphenylporphyrins containing 40 or 80 boron atoms were synthesized and evaluated for their biodistribution and toxicity in EMT-6 tumor-bearing mice. Copper (II) meso-5,10,15,20-tetrakis[3-methoxy-4-(o-carboranylmethoxy)phenyl]porphyrin, 6, and copper (II) meso-5,10,15,20-tetrakis[3-hydroxy-4-(o-carboranylmethoxy)phenyl]porphyrin, 8, are B40 congeners with different lipophilicities, each less than their B80 congener, copper (II) meso-5,10,15,20-tetrakis[m-(3,5-di-o-carboranylmethoxybenzyloxy)phenyl]porphyrin, 18. Two days after the last of a series of i.p. injections in BALB/c mice bearing EMT-6 mammary tumors, a dose of 185 mg/kg 6 (54 mg/kg B) delivered over 3.5 times the concentration of boron to tumor (169 microg/g B) than did 118 mg/kg 8 (36 mg/kg B), which delivered 35 microg/g B, or 87 mg/kg 18 (30 mg/kg B), which delivered 46 microg/g B. The tumor-to-blood and tumor-to-brain boron concentration ratios at that time for all three porphyrins exceeded 80:1. Two days after the last injection, there resulted moderate thrombocytopenia that essentially disappeared two days later from 6 and 18, and mild leukocytosis from 6, 8, and 18, all of which were clinically inconsequential. Thus, 6 may rank among the most clinically promising carboranyl porphyrins ever made to deliver 10B to tumors for boron neutron-capture therapy (BNCT) that has also been tested for its toxicity in vivo.

Animals↗
PubMed2006

Role of connective tissue growth factor (CTGF) module 4 in regulating epithelial mesenchymal transition (EMT) in HK-2 cells.

BACKGROUND: Recent studies have suggested that connective tissue growth factor (CTGF) plays a key role in tissue fibrosis including renal scarring. While studies showed several forms of CTGF with 10-38 kDa in the body fluids, little is known about these small molecule species. We investigated the effect of a 10 kDa CTGF molecule consisting of module 4, on the epithelial mesenchymal transition (EMT) in human proximal tubular cell line (HK-2). METHODS: HK2 cells were cultured in DMEM medium. The response of cytokeratin (CK) and vimentin (VIM) mRNA and protein expression to the stimulation of rhCTGF(C) were observed by real-time PCR and immunocytochemistry. At the same time, the morphologic changes were observed by microscopy, and expression of alpha-smooth muscle actin (alpha-SMA) and fibronectin (FN) was detected by laser confocal microscope. These effects were compared with CTGF N-terminal [rhCTGF(N)], consisting of module 1-3, and observed in a condition with the addition of anti-CTGF antibody. RESULTS: RhCTGF(C) induced striking changes in epithelial cells, including changes in cellular morphology, loss of CK, gain VIM and alpha-SMA, and increased levels of fibronectin. Cocultured with anti-CTGF antibody could abrogate most of these effects, while cells treated with rhCTGF(N) showed no significant phenotypic changes comparing to control group. CONCLUSIONS: Our results suggest that module 4 could induce HK-2 cells EMT, whereas the residual fragment has no similar effect in spite of consisting of 3 modules of CTGF molecule.

Actins↗
PubMed1981

Electrophoretic mobility test (EMT): studies on lymphocyte response and mechanism of the test using a rat tumor model.

After incubation with an encephalitogenic factor from human (HEF) or rat (REF) brain, lymphocytes of Fischer 344 rats bearing a spontaneous mammary adenocarcinoma produced a soluble substance which reduced the mobility of tanned sheep red blood cells in the electrophoretic mobility test (EMT). For studying the kinetics of this lymphocyte response, 6 X 10(5) tumor cells were injected into the hind footpad. In correlation with time and tumor size, one was able to influence the appearance of metastases by amputation of the leg. As early as 16 hours after inoculation of tumor cells, sensitivity of lymphocytes against HEF and a KCl-extract of the tumor could be shown in the EMT. It decreased on days 2 and 5, but was still seen until the day of amputation. Rats without metastases showed sensitivity up to four weeks after amputation and then returned to normal levels. Rats with metastases showed sensitivity until death at about seven weeks later. With the use of Amicon membranes, Sephadex G-50, and ion-exchange chromatography, a protein could be isolated from human basic myelin extract with a molecular weight of about 16,000-20,000 daltons. It had no direct influence on the EIC by itself, but after incubation with lymphocytes from tumor-bearing rats it evoked the production of a slowing substance. Using Sephadex G-100, the slowing substance appeared in the region in front of BSA indicating a molecular weight of greater than 80,000 daltons. It was heat-stable for 30 min at 56 degrees C and was sensitive to trypsin.

Adenocarcinoma↗
PubMed1981

Congruency in physician-EMT assessment.

The transport of 488 patients by Department of Health and Hospital (DHH) ambulances during a three-week period was studied in an effort to determine the operational effectiveness of the Point of Entry (POE) Plan in Boston. The Plan is based on an assumption that EMTs can correctly assess patients' medical problems. Comparisons of the physicians' diagnoses and the EMTs' assessments were made to establish the level of congruency for each of the seven care categories delineated. Several secondary univariate analyses were performed for their association with accuracy in patient assessment. Appropriate ambulance utilization also was reviewed based on the diagnosis and the circumstances of the incident. Findings supported the effectiveness of the assessment and transportation of emergency patients in the Boston emergency medical services system.

Acute Disease↗
PubMed1976

Can the well trained EMT-paramedic maintain skills and knowledge?

Reliable criteria exist to assess emergency medical technician (EMT)-paramedic team and individual competence. In addition to written and oral examiniation, reliable individual manipulative skill data, and on-scene observation of emergency care by responsible physicians, EMT-paramedic team competence may be assessed by using tracer conditions that reflect patient outcome converted to a standard base population per unit time. Using standard definitions of "resuscitation" and "save," and the pulseless, nonbreathing patient as tracer, 75 (31%) of 241 patients were resuscitated and 34 (14%) were saved. Team manipulative skill competence may also be determined by an appropriate tracer.

Allied Health Personnel↗
PubMed1981

Cell electrophoresis for diagnostic purposes. I. Diagnostic value of the electrophoretic mobility test (EMT) for the detection of gynaecological malignancies.

Lymphocytes from 278 gynaecological patients (100 controls and 178 patients with a malignant condition) have been investigated for their response to encephalitogenic factor, cancer basic protein, and KCl extract of adenocarcinoma of the body of the uterus as "antigens", using tanned sheep erythrocytes ETS as indicator particles in the electrophoretic mobility test (EMT). Electrophoretic mobility was measured with a Zeiss cytopherometer. The study was split into three test series producing in the cancer group 66% correct positive test results (34% false negatives) and in the control group 83% correct negative results (17% false positives). Consequently, with the instrumentation used, EMT is, at least in our hands, not sufficiently reliable for the diagnosis of cancer.

Adenocarcinoma↗
PubMed2005

Rac1b and reactive oxygen species mediate MMP-3-induced EMT and genomic instability.

The tumour microenvironment can be a potent carcinogen, not only by facilitating cancer progression and activating dormant cancer cells, but also by stimulating tumour formation. We have previously investigated stromelysin-1/matrix metalloproteinase-3 (MMP-3), a stromal enzyme upregulated in many breast tumours, and found that MMP-3 can cause epithelial-mesenchymal transition (EMT) and malignant transformation in cultured cells, and genomically unstable mammary carcinomas in transgenic mice. Here we explain the molecular pathways by which MMP-3 exerts these effects: exposure of mouse mammary epithelial cells to MMP-3 induces the expression of an alternatively spliced form of Rac1, which causes an increase in cellular reactive oxygen species (ROS). The ROS stimulate the expression of the transcription factor Snail and EMT, and cause oxidative damage to DNA and genomic instability. These findings identify a previously undescribed pathway in which a component of the breast tumour microenvironment alters cellular structure in culture and tissue structure in vivo, leading to malignant transformation.

Alternative Splicing↗