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 arbejde blev finansieret delvist af DK089016 og L30 RR030244 (MAC), CA153036 (AS), og P30-DK52574 (til Washington University Digestive Diseases Research Core). AIT var en Howard Hughes Medical Institute Medical Research Training Fellow.

1. Colitis-associeret cancerinduktion <ol><li> Braklægning bure af køn og alder-matchede 6-8 uger gamle mus, der skal anvendes til forsøg og kontrol grupper. Mus kan individuelt mærket med hale markeringer eller øreclips. </li><li> På dag 0, optage baseline vægt og injicere hver mus intraperitonealt (IP) med 10 mg / kg af AOM arbejdsopløsning (1 mg / ml i isotonisk saltvand, fortyndet fra 10 mg / ml stamopløsning i dH2O opbevares ved -20 grader C) . Baseret på erfaring, kan denne dosis justeres mellem 7-14 m...

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Behandling af mus med AOM og DSS hurtigt og effektivt modeller human colitis-associeret cancer. Hypoteser vedrørende arvelige faktorer, colitis-associerede cancere er let studeret med genetisk modificerede mus. 13, 16 Alternativt kan virkningen af farmakologiske mål i colitis-associeret cancer blive undersøgt ved anvendelse af vild-type mus. Mens denne model er stærkt værdsat af dem interesseret i studiet af colontumor udvikling i fastsættelsen af ​​inflammation, findes ...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalognummer </td><td> Kommentarer </td></tr><tr><td> C57BL/6J-mus </td><td> Jackson Laboratory </td><td> 000.664 </td><td></td></tr><tr><td> Azoxymethane (AOM) </td><td> Sigma Aldrich </td><td> A5486-100mg </td><td> Stamopløsning: fortynd til 10 mg / ml i destilleret vand holdes ved -20 ° C i 0,5 til 1 ml alikvoter. Arbejde opløsning: lager til 1 mg / ml i isotonisk (0,9%) saltvand </td></tr><tr><td> Dextransulfat Natrium (DSS) </td><td> TDB Consultancy </td><td> DB001 </td><td> MW 40 kDa (36-50 kDa hidrørende fra andre kilder er acceptable, det samme parti bør anvendes til et enkelt eksperiment) 6 </td></tr><tr><td> Coloviewminiendoscopic-system </td><td> Karl Storz </td><td> Multiple </td><td> Se Becker et al. For detaljeret forklaring af udstyr og opsætning. 11 </td></tr><tr><td> TPP Rapid FilterMax 500 ml Flaske-Filter, 0,22 um PES </td><td> Midwest Scientific </td><td> TP99500 </td><td> Enhver standard vævskultur filter er acceptabel </td></tr><tr><td> Ethylalkohol 200 Proof ASC / USP </td><td> Farmako-AAPER (eller andre) </td><td> 11ACS200 </td><td> Fortyndes til 70% i destilleret vand </td></tr><tr><td> Isofluran, USP </td><td> Butler Animal Health Supply </td><td> 4029405 </td><td> Placer mus i glas krukke med gaze eller en lille klud vædet i narkose </td></tr><tr><td> 18G Straight gavage Needle </td><td> Braintree Scientific </td><td> N-008 </td><td></td></tr><tr><td> Phosphatbufret saltvand (PBS) </td><td> SigmaAldrich </td><td> P5493 </td><td> Fortyndes til 1X (0,01 M) i destilleret vand </td></tr><tr><td> Kold Bakke (Tissue Tek II Cold Plate) </td><td> Fisher Scientific </td><td> NC9491941 </td><td> Opbevar ved -20 ° C </td></tr><tr><td> ImageJ Software </td><td> NIH (gratis download) </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> Fortyndes til 10% i PBS </td></tr><tr><td> BD Bacto Agar </td><td> Fisher Scientific </td><td> DF0140-01-0 </td><td> Brug varmeplade at oprette 2% opløsning i destilleret vand </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; Buede punkter forebygge skader på tyktarmen under åbning. </td></tr><tr><td> Alcian Blue 8GX (pulver) </td><td> Sigma Aldrich </td><td> A5268 </td><td> Der tilsættes 1 g pulver til 100 ml 3% eddikesyre (3 ml iseddike + 97 ml destilleret vand) </td></tr><tr><td> 1 ml tuberkulin sprøjte med påmonteret 26 G x 3/8 i intradermal bevel nål </td><td> BD </td><td> 305.946 </td><td> Til injektion af AOM </td></tr></tbody></table>

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

Personer med inflammatorisk tarmsygdom (IBD), såsom Crohns sygdom (CD) eller colitis ulcerosa (UC) er i øget risiko for at udvikle kolorektal cancer (CRC) over raske personer. Denne risiko er proportional med varigheden og omfanget af sygdom, med en kumulativ incidens så høj som 30% hos individer med langvarige UC med udbredt colon engagement. 1 Colonic dysplasi i IBD og colitis associeret cancer (CAC) menes at udvikle sig som et resultat af gentagne cykler af epitelial-celle skade og reparation, mens disse celler bades i en kronisk inflammatorisk cytokin miljø. 2 Mens spontane og colitis-associerede cancere deler kvalitet er adenocarcinomer er sekvensen af de underliggende molekylære begivenheder menes at være anderledes. 3. Denne skelnen argumenterer behovet for specifikke dyremodeller for CAC. Adskillige musemodeller øjeblikket findes for studiet af CAC. Dextransulfat Sodium (DSS), et middel med direkte toksiske virkninger på tyktarmsepitel, kan indgives i drikkevand til mus i flere cyklusser for at skabe en kronisk inflammatorisk tilstand. Med tilstrækkelig varighed, vil nogle af disse mus udvikler tumorer. Fire tumorudviklingen er fremskyndes i denne model hvis den gives i en pro-kræftfremkaldende indstilling. Disse omfatter mus med genetiske mutationer i tumorigenese pathways (APC, p53, MSH2), såvel som mus forbehandlet med genotoksiske midler (azoxymethane [AOM], 1,2-dimethylhydrazin [DMH]). Fem Kombinationen af ​​DSS med AOM som en model for colitis associeret cancer har vundet popularitet for sin reproducerbarhed, styrke, lav pris, og brugervenlighed. Selv om de har en fælles mekanisme, der AOM vist sig at være mere potente og stabile i opløsning end DMH. Mens tumorudvikling hos andre modeller kræver almindeligvis flere måneder, mus injiceret med AOM og efterfølgende behandlet med DSS udvikle passende tumorer i ens lidt som 7-10 uger. 6, 7 Endelig AOM og DSS kan administreres til mus af alle genetiske baggrund (knock out, transgene osv.) uden krydsning til en specifik tumorigen stamme. Her viser vi en protokol for inflammation drevet colon tumorigenese i mus under anvendelse af en enkelt injektion af AOM efterfulgt af tre syv-dages cyklusser af DSS over en 10 ugers periode. Denne model fremkalder tumorer med histologiske og molekylære forandringer ligger tæt op ad dem, der forekommer i human CAC og giver en meget værdifuld model til undersøgelse af onkogenese og chemoprevention i denne sygdom. 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 protokol, hvor administration af genotoksisk stof azoxymethane (AOM) efterfulgt af tre cyklusser af det pro-inflammatoriske middel dextransulfat-natrium (DSS) hurtigt og konsekvent frembringer colontumorer hos mus med morfologiske og molekylære lighed med dem, der ses i human colitis -associeret cancer.

Modeling Colitis-Associated Cancer med Azoxymethane (AOM) og dextransulfat 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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