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

Questa ricerca è finanziata dalla vescica AACR Henry Shepard Cancer Research Grant (09-60-30-LI).

Procedure che coinvolgono soggetti animali sono stati approvati dal Animal Care istituzionale e Comitato uso (IACUC) presso l&#39;University of California, San Francisco. 1. Preparazione cellulare <ol><li> La linea di carcinoma della vescica umana cellule KU-7-luc2-GFP (Caliper Life Sciences, Hopkinton, MA) viene coltivato in RPMI-1640 integrato con 10% siero fetale bovino e 50 ug / ml di gentamicina. </li><li> Le colture vengono mantenute a 37 ° C, 5% CO 2 con mezzi cambia ogn...

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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 presentiamo un protocollo per l&#39;istituzione di un modello murino ortotopica tumore della vescica xenotrapianto seguito da un trattamento intravescicale dei tumori con sarna LNP formulato che bersaglia il promotore di CIP1/WAF1 p21 (p21) per l&#39;attivazione della trascrizione. 2, 3 I tumori risultanti possono essere confermata e monitorata con immagini e bioluminescenti ultrasuoni vescica. Modelli ortotopica può essere superiore a intraperitoneale, sottocutanea, endovenosa o modelli fornen...

<table border="1"><tbody><tr><td> Nome del reattivo </td><td> Azienda </td><td> Numero di catalogo </td><td> Commenti (opzionale) </td></tr><tr><td> KU-7-luc-GFP le cellule </td><td> Pinza </td><td> 128091 </td><td></td></tr><tr><td> timici topi nudi (nu / nu) </td><td> Simonsen Laboratories </td><td> 6-7 settimane </td><td> Sim: (NCR) nu / nu fisol </td></tr><tr><td> Ultrasuoni unità </td><td> Visualsonics </td><td> Vevo 770 </td><td> RMV-704 sonda </td></tr><tr><td> Bioluminescenza unità </td><td> Pinza </td><td> IVIS Spectrum </td><td></td></tr><tr><td> Exel safelet catetere </td><td> Pescatore </td><td> 14-841-21 </td><td> 24G X ¾ &quot; </td></tr><tr><td> Di nitrato d&#39;argento </td><td> Sigma </td><td> S8157 </td><td></td></tr><tr><td> Hamilton siringa </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> Tabella 1. Reagenti e attrezzature specifiche.

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

Presentiamo un nuovo metodo per il trattamento del cancro della vescica con attivante RNA via endovescicale consegnato piccolo (sarna) in un modello murino ortotopico eterotrapianto tumore della vescica. Il modello murino è stabilito dal cateterismo uretrale sotto anestesia generale per via inalatoria. Combustioni chimiche viene quindi introdotto alla mucosa della vescica utilizzando intravescicale soluzione di nitrato d&#39;argento di distruggere lo strato di glicosaminoglicani vescica e permette alle cellule di allegare. Dopo alcuni lavaggi con acqua sterile, cancro della vescica umana KU-7-luc2-GFP cellule sono instillati attraverso il catetere in vescica di sosta per 2 ore. Successiva crescita dei tumori vescicali è confermata e monitorata da ecografia vescicale in vivo e di imaging bioluminescente. I tumori vengono poi trattati per via endovescicale con sarna formulata in nanoparticelle lipidiche (LNPs). La crescita del tumore è monitorata con gli ultrasuoni e la bioluminescenza. Tutte le fasi di questa procedura sono illustrati nel video allegato.

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

Stabilire un modello di tumore della vescica ortotopica per valutare gli effetti antitumorali di sarna via endovescicale consegnate e monitoraggio della crescita tumorale con l&#39;ecografia e imaging bioluminescente.

Un modello ortotopico tumore della vescica e la valutazione del trattamento intravescicale sarna

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

Research

JoVE Journal

Medicine

An Orthotopic Model of Murine Bladder Cancer

Un modello ortotopico di cancro alla vescica Murine

In this straightforward procedure, bladder tumors are established in female C57 mice through the use of catheterization, local cauterization, and ...

Ricerca

Medicina

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Multi-fotone Imaging di invasione delle cellule tumorali in un modello di mouse ortotopico di carcinoma orale a cellule squamose

Loco-regional invasion of head and neck cancer is linked to metastatic risk and presents a difficult challenge in designing and implementing patient ...

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Glioblastoma multiforme (GBM) are the most common and aggressive adult primary brain tumors1. In recent years there has been substantial progress in the ...

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Un modello di mouse ortotopico di anaplastico della tiroide Carcinoma

Several types of animal models of human thyroid carcinomas have been established, including subcutaneous xenograft and orthotopic implantation of cancer ...

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Un ortotopico cancro della vescica Modello di Gene Studi di consegna

Bladder cancer is the second most common cancer of the urogenital tract and novel therapeutic approaches that can reduce recurrence and progression are ...

An Orthotopic Murine Model of Human Prostate Cancer Metastasis

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Minimally Invasive Establishment of Murine Orthotopic Bladder Xenografts

Mini-Invasiva Istituzione di murini ortotopico vescica Xenotrapianti

Orthotopic bladder cancer xenografts are the gold standard to study molecular cellular manipulations and new therapeutic agents&#160;in vivo. Suitable ...

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Specialized immune cells that infiltrate the tumor microenvironment regulate the growth and survival of neoplasia.&#160; Malignant cells must elude or ...

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Evaluation of the Efficacy of the H. pylori Protein HP-NAP as a Therapeutic Tool for Treatment of Bladder Cancer in an Orthotopic Murine Model

Valutazione dell&#39;efficacia della<em&gt; H. pylori</em&gt; Proteine ​​HP-NAP come strumento terapeutico per il trattamento del cancro della vescica in un ortotopico Modello murino

Bladder cancer is one of the most common malignancies of the urogenital tract. Intravesical injection of Bacillus Calmette-Gu&#233;rin (BCG) is the gold ...

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The majority of cancer-related deaths are caused by the metastasis of the cancer rather than the primary tumor itself. Yet, the underlying mechanisms of ...