Name: modeling colitis-associated cancer with azoxymethane (aom) and dextran sulfate sodium (dss)
Uploaded: 2012-09-11T13:00:00
Description: Watch this Scientific Journal Video about Modeling Colitis-Associated Cancer with Azoxymethane AOM and Dextran Sulfate Sodium DSS at JoVE.com

Dette arbeidet ble finansiert delvis av DK089016 og L30 RR030244 (MAC), CA153036 (AS), og P30-DK52574 (til Washington University Digestive Diseases Forskning Core). AIT var en Howard Hughes Medical Institute Medical Research Training Fellow.

1. Kolitt-assosiert induksjon av kreft <ol><li> Sett til side bur av kjønn og alder-matchet 6-8 uker gamle mus som skal brukes til eksperimentelle og kontroll grupper. Mus kan være individuelt merket med hale markeringer eller øre klipp. </li><li> På dag 0, posten baseline vekter og injisere hver mus intraperitonealt (IP) med 10 mg / kg av AOM arbeidsoppløsning (1 mg / ml i isotonisk saltvann, fortynnet fra 10 mg / ml stamoppløsning i dH2O holdt ved -20 ° C) . Basert på erfaring, kan denne dosen justeres mello...

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Behandling av mus med AOM og DSS raskt og effektivt modeller humant kolitt-assosiert kreft. Hypoteser om arvelige faktorer som bidrar til kolitt-assosierte kreft kan lett studert med genmanipulerte mus. 13, 16 Alternativt kan effekten av farmakologisk mål i kolitt-assosiert kreft bli studert ved anvendelse villtype mus. Selv om denne modellen er høyt verdsatt av de som er interessert i studiet av tykktarmen tumorutviklingen i innstillingen av betennelse, begrensninger eksisterer...

<table border="1"><tbody><tr><td> Navnet på reagensen </td><td> Selskapet </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> C57BL/6J Mus </td><td> Jackson Laboratory </td><td> 000664 </td><td></td></tr><tr><td> Azoxymethane (AOM) </td><td> Sigma Aldrich </td><td> A5486-100mg </td><td> Stamoppløsning: fortynn til 10 mg / ml i destillert vann for å bli holdt ved -20 ° C som 0,5 til 1 ml alikvoter. Arbeide løsning: fortynne lager til 1 mg / ml i isotonisk (0,9%) saltvann </td></tr><tr><td> Dekstransulfat Natrium (DSS) </td><td> TdB Consultancy </td><td> DB001 </td><td> MW 40 kDa (36-50 kDa preparater fra andre kilder er akseptable; Samme mye bør brukes for ett enkelt eksperiment) 6 </td></tr><tr><td> Coloviewminiendoscopic system </td><td> Karl Storz </td><td> Multiple </td><td> Se Becker et al. For detaljert forklaring av utstyr og oppsett. 11 </td></tr><tr><td> TPP Rapid FilterMax 500 ml flaske-filter, 0,22 mikrometer PES </td><td> Midtvesten Scientific </td><td> TP99500 </td><td> Noen standard vevskultur filter er akseptabelt </td></tr><tr><td> Etylalkohol 200 Proof ASC / USP </td><td> Farmako-AAPER (eller andre) </td><td> 11ACS200 </td><td> Fortynn til 70% i destillert vann </td></tr><tr><td> Isofluran, USP </td><td> Butler Animal Health forsyning </td><td> 4029405 </td><td> Sted mus i glasskrukke med gasbind eller en liten klut dyppet i narkose </td></tr><tr><td> 18G Rett gavage Needle </td><td> Braintree Scientific </td><td> N-008 </td><td></td></tr><tr><td> Fosfatbufret saltoppløsning (PBS) </td><td> SigmaAldrich </td><td> P5493 </td><td> Fortynn til 1X (0,01 M) i destillert vann </td></tr><tr><td> Cold skuff (Tissue Tek II Cold Plate) </td><td> Fisher Scientific </td><td> NC9491941 </td><td> Oppbevares ved -20 ° C </td></tr><tr><td> ImageJ Software </td><td> NIH (gratis nedlasting) </td><td></td><td> <a href="http://rsbweb.nih.gov/ij/" target="_blank">http://rsbweb.nih.gov/ij/</a> </td></tr><tr><td> Formaldehyd (37%) </td><td> Fisher Scientific </td><td> F79-500 </td><td> Fortynn til 10% i PBS </td></tr><tr><td> BD Bacto Agar </td><td> Fisher Scientific </td><td> DF0140-01-0 </td><td> Bruk kokeplate å opprette 2% løsning i destillert vann </td></tr><tr><td> Miltex Eye Dressing tang </td><td> MedPlus Inc. </td><td> 18-780 </td><td></td></tr><tr><td> Miltex Eye Saks </td><td> MedPlus Inc. </td><td> 18-1430 </td> &lt;td&gt; kurvepunkter forhindre skade på tykktarmen ved åpning. </td></tr><tr><td> Alcian Blå 8GX (pulver) </td><td> Sigma Aldrich </td><td> A5268 </td><td> Tilsett 1 g pulver til 100 ml 3% eddiksyre (3 ml iseddik + 97 ml destillert vann) </td></tr><tr><td> 1 ml tuberculin sprøyte med vedlagte 26 G x 3/8 i intradermal skråkant nål </td><td> BD </td><td> 305946 </td><td> For injeksjon av AOM </td></tr></tbody></table>

modeling colitis-associated cancer with azoxymethane (aom) and dextran sulfate sodium (dss)

Personer med inflammatorisk tarmsykdom (IBD), slik som Crohns sykdom (CD) eller ulcerøs kolitt (UC) har økt risiko for å utvikle kolorektal kreft (CRC) over friske individer. Denne risikoen er proporsjonal med varigheten og omfanget av sykdommen, med en kumulativ insidens så høyt som 30% i individer med langvarig UC med utbredt colonic engasjement. 1 Colonic dysplasi i IBD og kolitt assosiert kreft (CAC) antas å utvikle seg som et resultat av gjentatte sykluser av epitelcelleopprinnelse skade og reparasjon mens disse cellene er badet i en kronisk inflammatorisk cytokin miljø. 2 Mens spontane og kolitt-assosiert kreft dele kvaliteten av å være adenocarcinomas, blir sekvensen av underliggende molekylære hendelser antas å være annerledes. 3 Denne skillet argumenterer behovet for spesifikke dyremodeller av CAC. Flere musemodeller dag eksisterer for studiet av CAC. Dekstransulfat Sodium (DSS), en agent med direkte toksiske effekter på tykktarmsområdet epitel, kan administreres i drikkevann til mus i flere sykluser for å skape en kronisk inflammatorisk tilstand. Med tilstrekkelig varighet, vil noen av disse musene utvikler svulster. 4 Tumor utvikling er fremskyndet i denne modellen hvis administreres i en pro-kreftfremkallende setting. Disse inkluderer mus med genetiske mutasjoner i tumorigenesis trasé (APC, p53, Msh2) samt mus forbehandlet med genotoksiske midler (azoxymethane [AOM], 1,2-dimethylhydrazine [DMH]). 5 Kombinasjonen av DSS med AOM som modell for kolitt assosiert kreft har vunnet popularitet for sin reproduserbarhet, styrke, lav pris, og brukervennlighet. Selv om de har en felles mekanisme, har AOM blitt funnet å være mer potent og stabil i løsning enn DMH. Mens tumor utvikling i andre modeller generelt krever flere måneder, mus injisert med AOM og deretter behandlet med DSS utvikle adekvate svulster i ens lite som 7-10 uker. 6, 7 Endelig AOM og DSS kan administreres til mus av noen genetisk bakgrunn (knock out, transgene, etc.) uten kryss-avl til en bestemt svulstfremkallende belastning. Her viser vi en protokoll for betennelse-drevet colonic tumorigenesis i mus som benytter en enkelt injeksjon av AOM etterfulgt av tre syv-dagers sykluser av DSS over en 10 ukers periode. Denne modellen induserer svulster med histologiske og molekylærbiologiske forandringer tett likner de som forekommer i menneskelig CERT og gir en svært verdifull modell for studiet av oncogenesis og Kjemoprevensjon i denne sykdommen. 8

Watch this Scientific Journal Video about Modeling Colitis-Associated Cancer with Azoxymethane AOM and Dextran Sulfate Sodium DSS at JoVE.com

Modeling Colitis-Associated Cancer with Azoxymethane AOM and Dextran Sulfate Sodium DSS

Vi viser en protokoll der administrasjon av gentoksisk stoff azoxymethane (AOM) etterfulgt av tre sykluser av pro-inflammatorisk agent dekstransulfat natrium (DSS) raskt og konsekvent genererer tykktarmskreft hos mus med morfologiske og molekylære likheter til de som er sett i menneskelig kolitt -assosiert kreft.

Modellering Kolitt-Associated Cancer med Azoxymethane (AOM) og dekstransulfat Natrium (DSS)

Individuals with inflammatory bowel disease (IBD), such as Crohn's disease (CD) or ulcerative colitis (UC) are at increased risk of developing colorectal ...

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Modeling Colitis-Associated Cancer with Azoxymethane (AOM) and Dextran Sulfate Sodium (DSS)

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