原位膀胱肿瘤模型和评价治疗膀胱萨纳
Summary
建立原位膀胱肿瘤模型来评估膀胱内交付萨纳和超声监测肿瘤的生长和发光成像的抗肿瘤作用。
Abstract
我们提出了一个新颖的方法治疗膀胱癌与膀胱内交付的小活化RNA（萨尔纳）的原位移植瘤小鼠膀胱肿瘤模型。小鼠模型的建立吸入全身麻醉下导尿。化学烧伤，然后介绍了使用膀胱内的硝酸银溶液膀胱粘膜破坏膀胱聚糖层，使细胞附着。用无菌水，人膀胱癌KU-7-luc2-GFP细胞经过几个清洗灌输通过导管进入膀胱停留2个小时。被确认和监测膀胱肿瘤的后续增长在体内的膀胱超声和生物发光成像。萨纳脂质纳米粒（岭澳核电站）制定的肿瘤治疗膀胱内。肿瘤的生长监测与超声和发光。此过程的所有步骤都体现在影随行。
Protocol
已批准制度的动物护理和使用委员会（IACUC），旧金山加州大学动物学科，涉及的程序。 1。细胞的制备是生长在人类膀胱癌细胞株KU-7-luc2-GFP（卡尺生命科学，霍普金顿，马）的RPMI-1640含10％胎牛血清和50微克/毫升的庆大霉素补充媒体。 文化是保持在37℃，5％，与媒体的CO 2的变化，每两至三天。 胰蛋白酶消化收获细胞，并计算使用血球。一个单?…
Discussion
随后制定的LNP萨纳转录激活目标为P21 CIP1/WAF1（P21）启动肿瘤灌注治疗鼠标原位移植膀胱肿瘤模型的建立，我们提出了一个协议。2，由此产生的肿瘤可以确认和监测生物发光成像和膀胱超声。原位模型可能优于通过腹腔，皮下或静脉注射模型提供了一个环境，更接近内源性膀胱癌的尿液和尿路上皮。各种原位小鼠膀胱肿瘤已建立了直接的膀胱壁注射以及灌注4-8肿瘤灌注，?…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项研究是由亨利·谢泼德的AACR膀胱癌的研究资助（09-60-30-李）。
Materials
| Name of the reagent | Company | Catalogue number | Comments (optional) |
| KU-7-luc-GFP cells | Caliper | 128091 | |
| thymic nude mice ( nu/nu) | Simonsen Laboratories | 6-7 weeks old | Sim:(NCr) nu/nu fisol |
| Ultrasound unit | Visualsonics | Vevo 770 | RMV-704 probe |
| Bioluminescence unit | Caliper | IVIS Spectrum | |
| Exel safelet catheter | Fisher | 14-841-21 | 24G X ¾” |
| Silver nitrate | Sigma | S8157 | |
| Hamilton syringe | Hamilton company | 80301 | |
| LNP-saRNA | Alnylam Pharmaceuticals |
Table 1. Specific reagents and equipment.
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Tags
Orthotopic Bladder Tumor ModelIntravesical SaRNA TreatmentBladder Cancer TreatmentSmall Activating RNAOrthotopic Xenograft Mouse ModelUrethral CatheterizationInhaled General AnestheticChemical BurnIntravesical Silver Nitrate SolutionBladder Glycosaminoglycan LayerHuman Bladder Cancer CellsKU-7-luc2-GFP CellsCatheterIn Vivo Bladder UltrasoundBioluminescent ImagingLipid Nanoparticles (LNPs)Tumor Growth Monitoring
Cite This Article
Kang, M. R., Yang, G., Charisse, K., Epstein-Barash, H., Manoharan, M., Li, L. An Orthotopic Bladder Tumor Model and the Evaluation of Intravesical saRNA Treatment. J. Vis. Exp. (65), e4207, doi:10.3791/4207 (2012).