Name: an orthotopic bladder tumor model and the evaluation of intravesical sarna treatment
Uploaded: 2012-07-28T14:00:00
Description: Watch this Scientific Journal Video about An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment at JoVE.com

Denne forskning er finansieret af AACR Henry Shepard Blære Cancer Research Grant (09-60-30-LI).

Procedurer, der involverer dyr emner er blevet godkendt af Institutional Animal Care og Use Committee (IACUC) ved University of California, San Francisco. 1. Cellepræparat <ol><li> Den humane blærecancer cellelinie KU-7-luc2-GFP (Caliper Life Sciences, Hopkinton, MA) dyrkes i RPMI-1640 medium suppleret med 10% føtalt bovint serum og 50 pg / ml gentamicin. </li><li> Kulturerne holdes ved 37 ° C, 5% CO2 med mediet ændrer hver to til tre dage. </li><li> Cellerne blev høstet v...

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	<li>Semple, S. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Rational+design+of+cationic+lipids+for+siRNA+delivery.">Rational design of cationic lipids for siRNA delivery.</a> Nature. 28, 172-176 (2010).</li><li>Li, L. C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Small+dsRNAs+induce+transcriptional+activation+in+human+cells.">Small dsRNAs induce transcriptional activation in human cells.</a> Proceedings of the National Academy of Sciences of the United States of America. 103, 17337-17342 (2006).</li><li>Chen, Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Antitumor+effect+of+dsRNA-induced+p21(WAF1/CIP1)+gene+activation+in+human+bladder+cancer+cells.">Antitumor effect of dsRNA-induced p21(WAF1/CIP1) gene activation in human bladder cancer cells.</a> Molecular cancer therapeutics. 7, 698-703 (2008).</li><li>Chancellor, M. B. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Preliminary+results+of+myoblast+injection+into+the+urethra+and+bladder+wall:+a+possible+method+for+the+treatment+of+stress+urinary+incontinence+and+impaired+detrusor+contractility.">Preliminary results of myoblast injection into the urethra and bladder wall: a possible method for the treatment of stress urinary incontinence and impaired detrusor contractility.</a> Neurourology and urodynamics. 19, 279-287 (2000).</li><li>Hadaschik, B. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+validated+mouse+model+for+orthotopic+bladder+cancer+using+transurethral+tumour+inoculation+and+bioluminescence+imaging.">A validated mouse model for orthotopic bladder cancer using transurethral tumour inoculation and bioluminescence imaging.</a> BJU international. 100, 1377-1384 (2007).</li><li>Fu, C., Apelo, C. A., Torres, B., Thai, K. H., Hsieh, M. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Mouse+Bladder+Wall+Injection.">Mouse Bladder Wall Injection.</a> J. Vis. Exp. (53), e2523 (2011).</li><li>Dobek, G. L., Godbey, W. T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=An+Orthotopic+Model+of+Murine+Bladder.">An Orthotopic Model of Murine Bladder.</a> Cancer. J. Vis. Exp. (48), 2535 (2011).</li><li>Dinney, C. P. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Isolation+and+characterization+of+metastatic+variants+from+human+transitional+cell+carcinoma+passaged+by+orthotopic+implantation+in+athymic+nude+mice.">Isolation and characterization of metastatic variants from human transitional cell carcinoma passaged by orthotopic implantation in athymic nude mice.</a> The Journal of urology. 154, 1532-1538 (1995).</li></ol>

Vi præsenterer en protokol til etablering af en mus ortotopisk xenograft blæretumor model efterfulgt af intravesikal behandling af tumorer med LNP formuleret Särna der retter sig mod promotor af p21 CIP1/WAF1 (p21) for transkriptionsaktivering. 2, 3 De fremkomne tumorer kan være bekræftet og overvåges med bioluminiscerende billedbehandling og blære ultralyd. Orthotopisk model kan være overlegne i forhold til intraperitoneal, subkutan eller intravenøs modeller ved at tilvejebringe et miljø...

<table border="1"><tbody><tr><td> Navn på reagenset </td><td> Firma </td><td> Katalog nummer </td><td> Kommentarer (valgfri) </td></tr><tr><td> KU-7-luc-GFP-celler </td><td> Caliper </td><td> 128091 </td><td></td></tr><tr><td> thymiske nøgne mus (nu / nu) </td><td> Simonsen Laboratories </td><td> 6-7 uger gamle </td><td> Sim: (NCR) nu / nu fisol </td></tr><tr><td> Ultralydsenheden </td><td> Visualsonics </td><td> Vevo 770 </td><td> RMV-704-probe </td></tr><tr><td> Bioluminescence enhed </td><td> Caliper </td><td> IVIS Spectrum </td><td></td></tr><tr><td> Exel safelet kateter </td><td> Fisher </td><td> 14-841-21 </td><td> 24G x ¾ &quot; </td></tr><tr><td> Sølvnitrat </td><td> Sigma </td><td> S8157 </td><td></td></tr><tr><td> Hamilton sprøjte </td><td> Hamilton company </td><td> 80301 </td><td></td></tr><tr><td> LNP-Sarna </td><td> Alnylam Pharmaceuticals </td><td></td><td></td></tr></tbody></table> Tabel 1. Specifikke reagenser og udstyr.

an orthotopic bladder tumor model and the evaluation of intravesical sarna treatment

Vi præsenterer en ny fremgangsmåde til behandling af blærecancer med intravesikalt leveret lille aktiverende RNA (Särna) i en orthotopisk xenotransplantat muse blæren tumormodel. Den musemodel er etableret af urethral kateterisation under inhaleret narkose. Kemisk forbrænding indføres derefter til blæren ved hjælp intravesical sølvnitratopløsning at forstyrre blæren glycosaminoglycan lag og tillader celler at vedhæfte. Efter adskillige vaske med sterilt vand, human blærekræft KU-7-luc2-GFP-celler instilleret gennem kateteret ind i blæren for at hvile i 2 timer. Efterfølgende vækst af blæretumorer bekræftes og overvåges af in vivo blæren ultralyd og bioluminiscerende billedbehandling. De tumorer behandles derefter intravesikalt med Särna formuleret i lipidnanopartikler (LNPs). Tumor vækst overvåges med ultralyd og bioluminescens. Alle trin i denne procedure er vist i den ledsagende video.

Watch this Scientific Journal Video about An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment at JoVE.com

An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment

Etablering af en orthotopisk blæretumor model til at vurdere antitumor virkninger af intravesikalt leveret Särna og overvågning tumorvækst ved hjælp af ultralyd og bioluminiscerende billedbehandling.

En Orthotopisk blæretumor Model og evaluering af Intravesikal Sarna behandling

We present a novel method for treating bladder cancer with intravesically delivered small activating RNA (saRNA) in an orthotopic xenograft mouse bladder ...

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Medicine

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