chick-heart-invasion-assay-for-testing-invasiveness-cancer-cells

Chick Heart Invasion Assay for Testing the Invasiveness of Cancer Cells and the Activity of Potentially Anti-invasive Compounds

The goal of the chick heart assay is to offer a relevant organ culture method to study tumor invasion in three dimensions. The assay can distinguish ...

University of Ghent

To identify in human retinoblastoma and normal retinal tissue the type of cadherin, its relationship with cytoplasmic catenins, and its participation in invasion.

Structure and function of the N-cadherin/catenin complex in retinoblastoma.

Two effects of the citrus methoxyflavone tangeretin on cell-cell interactions are biologically relevant. Firstly, tangeretin upregulates the function of the E-cadherin/catenin complex in human MCF-7/6 breast carcinoma cells. This leads to firm cell-cell adhesion and inhibition of invasion in vitro. Secondly, tangeretin downregulates the interleukin-2 receptor on T-lymphocytes and natural killer cells. This leads to a decrease in the cytotoxic competence of these immunocytes against cancer cells. The second effect can become important when high doses of tangeretin are combined with adjuvant tamoxifen treatment for breast cancer. Experiments with nude mice bearing MCF-7/6 tumors showed that tangeretin given orally at high doses, abrogated the therapeutic suppression of tumor growth exerted by tamoxifen. No evidence for a tumor promoting effect of tangeretin by itself was found in these experiments. Tangeretin may be an interesting molecule for application in cases where immunosuppression could be clinically beneficial.

The citrus methoxyflavone tangeretin affects human cell-cell interactions.

A new and reliable HPLC method for the quantitation of daidzein, equol, and genistein in human breast tissue has been developed. The method was applied to biopsies from women undergoing breast reductions, who, prior to surgery, had ingested either a soy isoflavone preparation or a placebo tablet. The results were compared with data collected for urine and serum of the same subjects using standard methods. The limits of detection in the breast tissue homogenate were 24.7 nmol/l for daidzein, 148.0 nmol/l for equol, and 28.4 nmol/l for genistein (S/N of 3). The chromatographic limits of quantitation were 62.5 nmol/l for daidzein and genistein, and 125.0 nmol/l for equol, for which the accuracies were 86.0%, 83.6%, and 81.8%, respectively. The coefficients of variation of these measurements were all below 20% (11.1% for daidzein, 16.4% for genistein, and 13.2% for equol). The sample preparation comprised a concentration step and the absolute limits of quantitation were, therefore, 4.7 nmol/l, 18.8 nmol/l, and 0.94 nmol/l for daidzein and genistein, and 9.4 nmol/l, 37.5 nmol/l, and 1.9 nmol/l for equol in urine, serum, and breast tissue homogenate, respectively. Recoveries were between 70% (+/-5.6%) in breast tissue homogenate and 100% (+/-14.1%) in urine and serum for all three compounds. Equol (less than 1 micromol/l homogenate) was found to be the predominant phytoestrogen in breast tissue and its concentrations exceeded those in serum. The concentrations of phytoestrogens were at least 100-fold higher in urine than in serum and breast tissue.

Quantitation of soy-derived phytoestrogens in human breast tissue and biological fluids by high-performance liquid chromatography.

Une nouvelle m&eacute;thode HPLC fiable pour la quantification de la daidz&eacute;ine, g&eacute;nist&eacute;ine dans le tissu mammaire humain et &eacute;quol a &eacute;t&eacute; d&eacute;velopp&eacute;e. La m&eacute;thode a &eacute;t&eacute; appliqu&eacute;e aux biopsies de femmes subissant des r&eacute;ductions mammaires, qui, avant la chirurgie, avaient ing&eacute;r&eacute; une pr&eacute;paration d&#39;isoflavone de soja ou un comprim&eacute; placebo. Les r&eacute;sultats ont &eacute;t&eacute; compar&eacute;s avec les donn&eacute;es recueillies pour l&#39;urine et le s&eacute;rum des m&ecirc;mes sujets &agrave; l&#39;aide de m&eacute;thodes standards. Les limites de d&eacute;tection dans l&#39;homog&eacute;nat de tissus mammaires &eacute;taient 24,7 nmol/l pour la daidz&eacute;ine et 148,0 nmol/l pour l&#39;&eacute;quol 28,4 nmol/l pour la g&eacute;nist&eacute;ine (S/N 3). Limites de quantification par chromatographie en phase &eacute;taient 62,5 nmol/l pour la daidz&eacute;ine et la g&eacute;nist&eacute;ine et 125.0 nmol/l pour l&#39;&eacute;quol, pour lequel les pr&eacute;cisions &eacute;taient 86,0 % et 83,6 % 81,8 %, respectivement. Les coefficients de variation de ces mesures &eacute;taient toutes inf&eacute;rieures &agrave; 20 % (11,1 % de daidz&eacute;ine, 16,4 % pour la g&eacute;nist&eacute;ine et 13,2 % pour l&#39;&eacute;quol). La pr&eacute;paration de l&#39;&eacute;chantillon constitu&eacute; une &eacute;tape de concentration et les limites de quantification &eacute;taient, par cons&eacute;quent, 4,7 nmol/l, 18,8 nmol/l et 0,94 nmol/l daidz&eacute;ine et g&eacute;nist&eacute;ine, 9,4 nmol/l, 37,5 nmol/l et 1,9 nmol/l pour l&#39;&eacute;quol dans l&#39;urine, s&eacute;rum et homog&eacute;nat de tissu mammaire, respectivement. R&eacute;cup&eacute;ration &eacute;tait de 70 % (+/-5,6 %) dans l&#39;homog&eacute;nat de tissu mammaire &agrave; 100 % (+/-14,1 %) dans l&#39;urine et le s&eacute;rum pour les trois compos&eacute;s. &Eacute;quol (moins de 1 micromol/l homog&eacute;nat) se trouve &ecirc;tre le phytoestrog&egrave;ne pr&eacute;dominant dans le tissu mammaire et sa concentration d&eacute;passe le ceux dans le s&eacute;rum. Les concentrations de phytoestrog&egrave;nes &eacute;taient au moins 100 fois plus &eacute;lev&eacute;es dans l&#39;urine que dans le tissu de s&eacute;rum et de la poitrine.

Dosage des d&eacute;riv&eacute;s de soja phytoestrogens dans les liquides biologiques et les tissus du sein humain par chromatographie liquide &agrave; haute performance.

&Egrave; stato sviluppato un nuovo e affidabile metodo HPLC per la quantificazione di equol daidzeina e genisteina nel tessuto mammario umano. Il metodo &egrave; stato applicato a biopsie da donne in fase di riduzione del seno, che, prima dell&#39;intervento chirurgico, avevano ingerito una preparazione di isoflavoni di soia o una compressa di placebo. I risultati sono stati confrontati con i dati raccolti per urina e siero dei soggetti stessi utilizzando metodi standard. I limiti di rilevamento in omogeneizzato di tessuto del seno erano 24,7 nmol/l per la daidzeina, 148.0 nmol/l per equol e 28,4 nmol/l per la genisteina (S/N. 3). Cromatografici limiti di quantificazione sono stati 62,5 nmol/l per la daidzeina e genisteina e 125.0 nmol/l per equol, per cui le esattezze erano 86.0% 83,6% e 81,8%, rispettivamente. I coefficienti di variazione di queste misurazioni erano tutti sotto il 20% (11,1% per la daidzeina, 16,4% per genisteina e 13,2% per equol). La preparazione del campione comprendeva una fase di concentrazione e i limiti assoluti di quantizzazione erano, quindi, 4,7 nmol/l, 18,8 nmol/l e 0.94 nmol/l per la daidzeina e genisteina e 9,4 nmol/l, 37,5 nmol/l e 1,9 nmol/l per equol nell&#39;urina, siero e omogeneizzato di tessuto del seno, rispettivamente. Recuperi sono stati tra il 70% (+ 5,6%) in omogeneato di tessuto del seno e il 100% (+ 14,1%) nelle urine e siero per tutti e tre i composti. Equol (meno di 1 micromol/l omogeneizzato) &egrave; stato trovato per essere il Fitoestrogeno predominante nel tessuto del seno e le sue concentrazioni superato quelli nel siero. Le concentrazioni di fitoestrogeni sono stati almeno 100 volte pi&ugrave; elevate nelle urine che nel tessuto del siero e del seno.

Quantificazione dei fitoestrogeni derivati da soia in seno umano tessuto e biologici liquidi mediante cromatografia liquida ad alte prestazioni.

To investigate the role of N-cadherin and hepatocyte growth factor (HGF) in the invasion of collagen type I by human retinal pigment epithelial (RPE) cells.

Invasion of retinal pigment epithelial cells: N-cadherin, hepatocyte growth factor, and focal adhesion kinase.

Afin d&#39;&eacute;tudier le r&ocirc;le de facteur de croissance N-cadh&eacute;rine et des h&eacute;patocytes (HGF) lors de l&#39;invasion de collag&egrave;ne de type I par la r&eacute;tine humaine pigment des cellules &eacute;pith&eacute;liales (RPE).

Invasion des cellules &eacute;pith&eacute;liales pigmentaire r&eacute;tinien: N-cadh&eacute;rine, facteur de croissance des h&eacute;patocytes et kinase d&#39;adh&eacute;rence focale.

Per indagare il ruolo di N-caderina e del hepatocyte growth factor (HGF) nell&#39;invasione del collagene di tipo I da retina umana pigmento nelle cellule epiteliali (RPE).

Invasione delle cellule epiteliali del pigmento retinico: N-caderina, fattore di crescita degli epatociti e chinasi focale di adesione.

Invasion is the hallmark of malignant tumors, and is responsible for the bad prognosis of the untreated cancer patients. The search for anti-invasive treatments led us to screen compounds of different classes for their effect in an assay for invasion. Thirty-nine new compounds synthesized in the present study along with 56 already reported compounds belonging mainly to the classes of lactones, pyrazoles, isoxazoles, coumarins, desoxybenzoins, aromatic ketones, chalcones, chromans, isoflavanones have been tested against organotypic confronting cultures of invasive human MCF-7/6 mammary carcinoma cells with embryonic chick heart fragments in vitro. Three of them (a pyrazole derivative, an isoxazolylcoumarin and a prenylated desoxybenzoin) inhibited invasion at concentrations as low as 1 microM; instead of occupying and replacing the heart tissue within 8 days, the MCF-7/6 cells grew around the heart fragments and left it intact, when treated with these compounds. At the anti-invasive concentration of 1 microM, the three compounds did not affect the growth of the MCF-7/6 cells, as shown in the sulforhodamine B assay. Aggregate formation on agar was not stimulated by any of the three anti-invasive compounds, making an effect on the E-cadherin/catenin complex improbable. This is an invasion suppressor that can be activated in MCF-7/6 cells by a number of other molecules. Our data indicate that some polyphenolic and heterocyclic compounds are anti-invasive without being cytotoxic for the cancer cells.

Synthesis, characterization and in vitro anti-invasive activity screening of polyphenolic and heterocyclic compounds.

Invasion est la marque distinctive des tumeurs malignes et est responsable du mauvais pronostic des patients non trait&eacute;s du cancer. La recherche de traitements invahissant nous a conduit &agrave; des compos&eacute;s d&#39;&eacute;cran des diff&eacute;rentes classes pour leur effet lors d&#39;un test pour l&#39;invasion. Trente-neuf nouveaux compos&eacute;s synth&eacute;tis&eacute;s dans la pr&eacute;sente &eacute;tude ainsi que 56 d&eacute;j&agrave; rapport&eacute; compos&eacute;s appartenant principalement aux classes de lactones, pyrazoles, isoxazoles, coumarines, desoxybenzoins, c&eacute;tones aromatiques, chalcones, DIHYDROQUINOLINES, isoflavanones ont &eacute;t&eacute; test&eacute;s contre organotypique face aux cultures de cellules de carcinome mammaire MCF-7/6 humaines envahissantes avec fragments de coeur d&#39;embryon de poulet in vitro. Trois d&#39;entre eux (un d&eacute;riv&eacute; pyrazole, un isoxazolylcoumarin et un desoxybenzoin pr&eacute;nyl&eacute;s) inhibent invasion &agrave; des concentrations aussi faibles que 1 microM ; au lieu d&#39;occupation et remplacer le tissu de coeur dans les 8 jours, les cellules MCF-7/6 s&#39;est d&eacute;velopp&eacute;e autour des fragments de coeur et il a laiss&eacute; intact, avec ces compos&eacute;s. &Agrave; la concentration invahissant de 1 microM, les trois compos&eacute;s n&#39;a pas affect&eacute; la croissance des cellules MCF-7/6, comme indiqu&eacute; dans le test de sulforhodamine B. Formation agr&eacute;gation sur agar n&#39;a pas &eacute;t&eacute; stimul&eacute;e par l&#39;un des trois compos&eacute;s invahissant, ce qui rend un effet sur l&#39;ensemble de la E-cadh&eacute;rine/cat&eacute;nine improbable. Il s&#39;agit d&#39;un suppresseur d&#39;invasion qui peut &ecirc;tre activ&eacute; dans les cellules MCF-7/6, par un certain nombre d&#39;autres mol&eacute;cules. Nos donn&eacute;es indiquent que certains compos&eacute;s polyph&eacute;noliques et h&eacute;t&eacute;rocycliques sont invahissant sans &ecirc;tre cytotoxique pour les cellules canc&eacute;reuses.

Synth&egrave;se, caract&eacute;risation et d&eacute;pistage d&#39;activit&eacute; in vitro d&#39;invahissant de polyph&eacute;noliques et compos&eacute;s h&eacute;t&eacute;rocycliques.

Invasione &egrave; il segno distintivo dei tumori maligni ed &egrave; responsabile per la cattiva prognosi dei pazienti non trattati il cancro. La ricerca di trattamenti anti-invasive ci ha portato a composti di schermo di classi diverse per il loro effetto in un dosaggio per invasione. Trenta-nove nuovi composti sintetizzati nel presente studio insieme a 56 gi&agrave; segnalato composti appartenendo principalmente per le classi di lattoni, pyrazoles, isoxazoles, cumarine, desoxybenzoins, chetoni aromatici, calconi, chromans, isoflavanones sono stati testati contro organotypic affrontare culture di invasive cellule umane di carcinoma mammario MCF-7/6 con frammenti di cuore di pulcino embrionale in vitro. Tre di loro (un derivato di pyrazole, un isoxazolylcoumarin e un desoxybenzoin di prenylated) inibito invasione a concentrazioni bassi quanto 1 microM; invece di occupare e sostituendo il tessuto cardiaco entro 8 giorni, le cellule MCF-7/6 &egrave; cresciuta intorno i frammenti di cuore e lasciarono intatti, quando trattati con questi composti. La concentrazione del anti-invasive di 1 microM, i tre composti non hanno influito la crescita delle cellule MCF-7/6, come mostrato nel test cutaneo B. Formazione di aggregati su agar non era stimolato da uno qualsiasi dei tre composti anti-invasive, rendendo improbabile un effetto sul complesso E-caderina/catenina. Questo &egrave; un soppressore di invasione che pu&ograve; essere attivato in cellule MCF-7/6 da un certo numero di altre molecole. I nostri dati indicano che alcuni composti polifenolici ed eterociclici sono anti-invasive senza essere citotossiche per le cellule tumorali.

Sintesi, caratterizzazione e proiezione di attivit&agrave; anti-invasive in vitro di polifenoli e composti eterociclici.

Alkylphospholipids, such as the antitumor ether lipid 1-O-octadecyl-2-O-methylglycero-3-phosphocholine, modulate cancer cell invasion through changes in the adherens junction E-cadherin complex, a major organizer of epithelia. We wanted to know whether alkylphospholipids would also change tight junctions, molecular complexes that seal cell-to-cell contacts in polarised epithelia. Therefore, human colorectal cancer cell layers T84 were established in two-compartment culture chambers and the functional integrity of tight junctions was evaluated through their transepithelial electrical resistance. Incorporation of alkylphospholipids causes a rapid and reversible decrease of transepithelial electrical resistance. This decrease is due to an increased paracellular permeability and is temperature-independent. Unlike methyl-beta-cyclodextrin, alkyl-phospholipids do not specifically displace lipids from raft-like membrane domains. Nevertheless, alkylphospholipids change the detergent-solubility of zonula occludens-protein and occludin. Our data, together with the literature, indicate that non-toxic doses of alkylphospholipids affect more than one cell-cell adhesion complex, probably through their incorporation into the plasma membrane lipid bilayer.

Alkylphospholipids reversibly open epithelial tight junctions.

E-cadherin germline missense mutations have been shown to be responsible for significant loss of protein activity. A new cytoplasmic E-cadherin germline missense mutation (V832 M) was recently identified in a hereditary diffuse gastric cancer (HDGC) Japanese family. This E-cadherin mutant was cloned in a Chinese hamster ovary cell model system and functionally characterized, in terms of aggregation and invasion. Cells expressing the germline V832M mutant fail to aggregate and invade into collagen, supporting the pathogenic role of this germline missense mutation in gastric cancer. We also tested the ability of this mutation to activate the TCF-LEF trascriptional activity, in comparison with three other E-cadherin missense mutations (T340A, A634V and A617T), associated to loss of E-cadherin function. All the E-cadherin mutants reduced TCF-LEF activation to a similar extent as the wild-type protein, suggesting that the oncogenic effect of the E-cadherin mutants is unlikely to be transmitted through a beta-catenin-dependent activation of the WNT pathway.

The intracellular E-cadherin germline mutation V832 M lacks the ability to mediate cell-cell adhesion and to suppress invasion.

E-cadh&eacute;rine dans les cellules germinales des mutations faux-sens ont d&eacute;montr&eacute; d&#39;&ecirc;tre responsable de la perte significative de l&#39;activit&eacute; de la prot&eacute;ine. Une nouveau cytoplasmique E-cadh&eacute;rine dans les cellules germinales mutation faux-sens (V832 M) a &eacute;t&eacute; r&eacute;cemment identifi&eacute;e dans une famille japonaise de cancer gastrique diffus h&eacute;r&eacute;ditaire (HDGC). Ce mutant de E-cadh&eacute;rine a &eacute;t&eacute; clon&eacute; dans un syst&egrave;me de mod&egrave;le de cellules ovariennes hamster chinois et caract&eacute;ris&eacute; sur le plan fonctionnel, en termes d&#39;agr&eacute;gation et d&#39;invasion. Les cellules exprimant la lign&eacute;e germinale V832M mutant ne parviennent pas &agrave; agr&eacute;ger et envahir en collag&egrave;ne, appuyant le r&ocirc;le pathog&egrave;ne de cette mutation faux-sens de lign&eacute;e germinale dans le cancer gastrique. Nous avons aussi test&eacute; la capacit&eacute; de cette mutation pour activer la TCF-LEF trascriptional activit&eacute;, par rapport aux trois autres mutations de faux-sens E-cadh&eacute;rine (T340A, A634V et A617T), associ&eacute;e &agrave; la perte de fonction de E-cadh&eacute;rine. Tous les mutants de la E-cadh&eacute;rine r&eacute;duit activation TCF-LEF de mani&egrave;re similaire comme la prot&eacute;ine de type sauvage, ce qui sugg&egrave;re que l&#39;effet oncog&egrave;ne des E-cadh&eacute;rine mutants est peu susceptible d&#39;&ecirc;tre transmis par une activation de la b&ecirc;ta-cat&eacute;nine-d&eacute;pendante de la signalisation Wnt.

La mutation de cellules germinales E-cadh&eacute;rine intracellulaire V832 M n&#39;a pas la capacit&eacute; de m&eacute;diation adh&eacute;sion cellule-cellule et pour supprimer l&#39;invasion.

E-caderina germline mutazioni missenso hanno dimostrate di essere responsabile per la perdita significativa dell&#39;attivit&agrave; della proteina. Una nuova citoplasmatica E-caderina germline mutazione missenso (V832 M) &egrave; stata recentemente identificata in una famiglia giapponese cancro gastrico ereditario diffuso (HDGC). Questa mutante di E-caderina &egrave; stata clonata in un sistema modello di criceto cinese ovaia cella e funzionalmente caratterizzata, in termini di aggregazione e di invasione. Le cellule che esprimono il mutante V832M germinali non di aggregazione e invadono in collagene, sostenendo il ruolo patogeno di questa mutazione missenso germinale nel cancro gastrico. Abbiamo testato anche la capacit&agrave; di questa mutazione di attivare il TCF-LEF trascriptional attivit&agrave;, rispetto alle tre altre E-caderina mutazioni missenso (T340A, A634V e A617T), associata alla perdita della funzione di E-caderina. Tutti i mutanti di E-caderina ridotta attivazione TCF-LEF in misura analoga come la proteina wild-type, suggerendo che l&#39;effetto oncogenico dei mutanti E-caderina &egrave; improbabile da essere trasmessi attraverso un&#39;attivazione beta-catenina-dipendente del pathway WNT.

La mutazione di germline E-caderina intracellulare V832 M manca la capacit&agrave; di mediare l&#39;adesione cellula-cellula e di reprimere l&#39;invasione.

In subclones of the human colon cancer LoVo cell line, there is a reproducible spontaneous transition from an epithelioid (E) to a round (R) morphotype. The E to R transition is associated with increased cell growth, absence of E-cadherin-dependent compaction in a slow aggregation assay, loss of contact inhibition of motility and directional migration in a wound filling motility assay. Furthermore, none of the E subclones from LoVo was invasive into chick heart fragments. This is in contrast to the R subclones that were either nonadherent or adherent and invasive. Macroarray analysis demonstrated transcriptional downregulation of plakoglobin in R type LoVo cells and this was confirmed at the level of the mRNA by quantitative RT-PCR. Western blotting showed lower expression of all components of the E-cadherin/catenin complex in R subclones. Interestingly, treatment of R subclones with the demethylating agent 5-aza-2'-deoxycytidine resulted in restoration of the E morphotype, higher expression of E-cadherin, but not plakoglobin mRNA, and higher expression of E-cadherin and plakoglobin at the protein level.

Functional and molecular characterization of the epithelioid to round transition in human colorectal cancer LoVo cells.

Observations ant&eacute;rieures a sugg&eacute;r&eacute; que les opio&iuml;des modifient adh&eacute;rence cellule-substrat sur agar. Dans cette &eacute;tude, nous avons voulu &eacute;tudier si opio&iuml;des aussi nuire &agrave; l&#39;adh&eacute;rence cellulaire &agrave; des substrats biologiquement pertinentes, y compris les &eacute;l&eacute;ments de matrice et de la membrane basale interstitielles. Rein embryonnaire humain 293 cellules stablement transfect&eacute;s avec indicateur-&eacute;tiquet&eacute;es opiac&eacute;s-micro r&eacute;cepteur ont &eacute;t&eacute; utilis&eacute;es comme mod&egrave;le exp&eacute;rimental. Les cellules ont &eacute;t&eacute; cultiv&eacute;s sur culture de tissus plastique, collag&egrave;ne type I et IV, fibronectine, laminine et fragments de l&#39;amnios humain en absence ou en pr&eacute;sence de morphine. Cultures ont &eacute;t&eacute; immunomarqu&eacute;es avec antivinculin pour visualiser les contacts focaux. Cultures impr&eacute;gn&eacute;es de la morphine sur plastique de culture de tissus, les types de collag&egrave;ne I et IV et la fibronectine, mais pas sur laminine, couvert moins substrat que les cultures non trait&eacute;es ; les cellules individuelles sont difficiles &agrave; distinguer et leurs noyaux entassent de cultures par opposition &agrave; non trait&eacute;es. Les m&ecirc;mes effets ont &eacute;t&eacute; observ&eacute;s sur des fragments de l&#39;amnios humain : la morphine a chang&eacute; le morphotype des cultures sur le c&ocirc;t&eacute; stromal, mais pas sur le c&ocirc;t&eacute; de la membrane basale. Cultures qui ont &eacute;t&eacute; trait&eacute;s avec la morphine affichent moins de contacts focaux que les cultures non trait&eacute;es sur le collag&egrave;ne de type I, tandis que non trait&eacute;s et trait&eacute;s &agrave; la morphine les cultures &eacute;taient impossibles &agrave; distinguer sur la laminine.

Opio&iuml;des nuire &agrave; l&#39;adh&eacute;rence focale induite par le contact cellule-substrat.

Earlier observations suggested that opioids modify cell-substrate adhesion on agar. In this study, we wanted to investigate whether opioids also interfere with cell adhesion to biologically relevant substrates, including interstitial matrix and basement membrane components. Human embryonic kidney 293 cells stably transfected with FLAG-tagged micro-opioid receptor were used as an experimental model. The cells were cultured on tissue culture plastic, collagen types I and IV, fibronectin, laminin and human amnion fragments in the absence or presence of morphine. Cultures were immunolabelled with antivinculin to visualize focal contacts. Morphine-treated cultures on tissue culture plastic, collagen types I and IV and fibronectin, but not on laminin, covered less substrate than untreated cultures; individual cells were difficult to distinguish and their nuclei piled up in contrast to untreated cultures. The same effects were observed on human amnion fragments: morphine changed the morphotype of cultures on the stromal side, but not on the basement membrane side. Cultures that were treated with morphine displayed fewer focal contacts than untreated cultures on collagen type I, whereas untreated and morphine-treated cultures were indistinguishable on laminin.

Opioids affect focal contact-mediated cell-substrate adhesion.

Le osservazioni precedenti ha suggerito che gli oppioidi modificare adesione cellula-substrato su agar. In questo studio, abbiamo voluto indagare se gli oppioidi anche interferiscano con l&#39;adesione cellulare a substrati biologicamente rilevanti, compresi i componenti di matrice e membrana basale interstiziale. 293 Cellule Rene embrionale umano stabile transfected con bandiera-tagged recettore micro-oppioidi sono state utilizzate come modello sperimentale. Le cellule sono state coltivate su coltura di tessuto di plastica, collagene tipo I e IV, fibronectina, laminina e amnios umani frammenti nell&#39;assenza o la presenza di morfina. Le culture sono state immunolabelled con antivinculin per visualizzare i contatti focali. Culture morfina-trattato sulla coltura del tessuto di plastica, collagene tipo I e IV e fibronectina, ma non su laminina, coperto meno substrato di culture non trattate; singole celle erano difficili da distinguere e loro nuclei ammucchiati culture in contrasto a non trattate. Gli stessi effetti sono stati osservati su frammenti di amnios umani: morfina cambiato il morfotipo di culture sul lato stroma, ma non dal lato della membrana basale. Culture che sono state trattate con morfina visualizzato meno contatti focali di culture non trattate sul collagene di tipo I, mentre non trattati e trattati con morfina culture erano indistinguibili sulla laminina.

Gli oppiacei possono influenzare adesione focale mediato contatto cellula-substrato.

Tumors are microecosystems in which a continuous cross talk between cancer cells and host cells decides on the invasive behavior of the tumor cell population as a whole (Mareel et al., Encyclopedia of cancer, San Diego, CA, Academic Press, 1997). Both compartments secrete activating and inhibitory factors that modulate activities such as cell-extracellular matrix (ECM) interaction, cell-cell adhesion, remodeling of the ECM, and cell motility. For this reason, confrontations of cancer cells with a living normal host tissue in organ culture have been introduced by several groups: Wolff and Schneider in France (Wolff and Schneider, C R S Soc Biol (Paris) 151:1291-1292, 1957), Easty and Easty in the United Kingdom (Easty and Easty, Nature 199:1104-1105, 1963), and Schleich in Germany (Schleich et al., J Natl Cancer Inst 56:221-237, 1976). Embryonic chick heart fragments in organ culture maintain many histological features of their tissue of origin: They are composed of myocytes, fibroblasts, and endothelial cells, and their ECM contains fibronectin, laminin, and several collagen types. Moreover, the fragments remain contractile, and this activity allows the monitoring of their functional integrity during organ culture.

Marc M. Mareel

Department of Radiation Oncology and Experimental Cancer Research

University of Ghent

Chick Heart Invasion Assay for Testing the Invasiveness of Cancer Cells and the Activity of Potentially Anti-invasive Compounds

Marc M. Mareel

.css-o250i3{font-size:18px;font-family:Open Sans,sans-serif;line-height:21.6px;}Department of Radiation Oncology and Experimental Cancer Research

University of Ghent

Chick Heart Invasion Assay for Testing the Invasiveness of Cancer Cells and the Activity of Potentially Anti-invasive Compounds

Department of Radiation Oncology and Experimental Cancer Research