{"title":"Preclinical Models for Studying the Impact of Macrophages on Cancer Cachexia","authors":"Spas Dimitrov Markov, Daisy Gonzalez, Kamiya Mehla","doi":"10.1002/cpph.80","DOIUrl":null,"url":null,"abstract":"<p>Cancer-associated cachexia is defined by loss of weight and muscle mass, and by the potential loss of adipose tissue accompanied by insulin resistance and increased resting energy expenditure. Cachexia is most prevalent in pancreatic cancer, the third leading cause of cancer-related deaths. While various factors interact to induce cachexia, the precise mechanisms underlying this clinical condition are not fully understood. Clinically relevant animal models of cachexia are needed given the lack of standard diagnostic methods or treatments for this condition. Described in this article are in vitro and in vivo models used to study the role of macrophages in the induction of cachexia in pancreatic cancer. Included are procedures for isolating and culturing bone marrow−derived macrophages, harvesting tumor- and macrophage-derived conditioned medium, and studying the effect of conditioned medium on C2C12 myotubes. Also described are procedures involving the use of an orthotopic model of pancreatic cancer, including a method for examining skeletal muscle atrophy in this model. © 2020 Wiley Periodicals LLC.</p><p><b>Basic Protocol 1</b>: In vitro model of pancreatic tumor-induced cachexia using C2C12 cell lines (myotube model)</p><p><b>Support Protocol 1</b>: Molecular evaluation of cachectic markers in C2C12 myotubes using real-time PCR and immunoblotting</p><p><b>Basic Protocol 2</b>: In vivo model to study cachectic phenotype in pancreatic tumor-bearing mice</p><p><b>Support Protocol 2</b>: Evaluation of cachectic markers in the skeletal muscle of tumor-bearing mice</p>","PeriodicalId":10871,"journal":{"name":"Current Protocols in Pharmacology","volume":"91 1","pages":""},"PeriodicalIF":0.0000,"publicationDate":"2020-12-02","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"https://sci-hub-pdf.com/10.1002/cpph.80","citationCount":"3","resultStr":null,"platform":"Semanticscholar","paperid":null,"PeriodicalName":"Current Protocols in Pharmacology","FirstCategoryId":"1085","ListUrlMain":"https://onlinelibrary.wiley.com/doi/10.1002/cpph.80","RegionNum":0,"RegionCategory":null,"ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":null,"EPubDate":"","PubModel":"","JCR":"Q2","JCRName":"Pharmacology, Toxicology and Pharmaceutics","Score":null,"Total":0}
引用次数: 3
Abstract
Cancer-associated cachexia is defined by loss of weight and muscle mass, and by the potential loss of adipose tissue accompanied by insulin resistance and increased resting energy expenditure. Cachexia is most prevalent in pancreatic cancer, the third leading cause of cancer-related deaths. While various factors interact to induce cachexia, the precise mechanisms underlying this clinical condition are not fully understood. Clinically relevant animal models of cachexia are needed given the lack of standard diagnostic methods or treatments for this condition. Described in this article are in vitro and in vivo models used to study the role of macrophages in the induction of cachexia in pancreatic cancer. Included are procedures for isolating and culturing bone marrow−derived macrophages, harvesting tumor- and macrophage-derived conditioned medium, and studying the effect of conditioned medium on C2C12 myotubes. Also described are procedures involving the use of an orthotopic model of pancreatic cancer, including a method for examining skeletal muscle atrophy in this model. © 2020 Wiley Periodicals LLC.
Basic Protocol 1: In vitro model of pancreatic tumor-induced cachexia using C2C12 cell lines (myotube model)
Support Protocol 1: Molecular evaluation of cachectic markers in C2C12 myotubes using real-time PCR and immunoblotting
Basic Protocol 2: In vivo model to study cachectic phenotype in pancreatic tumor-bearing mice
Support Protocol 2: Evaluation of cachectic markers in the skeletal muscle of tumor-bearing mice
巨噬细胞对癌症恶病质影响的临床前模型研究
癌症相关恶病质的定义是体重和肌肉量的减少,以及伴随胰岛素抵抗和静息能量消耗增加的脂肪组织的潜在损失。恶病质在胰腺癌中最为普遍,胰腺癌是癌症相关死亡的第三大原因。虽然各种因素相互作用诱导恶病质,但这种临床状况的确切机制尚不完全清楚。由于缺乏标准的诊断方法或治疗方法,因此需要临床相关的恶病质动物模型。本文描述了用于研究巨噬细胞在胰腺癌恶病质诱导中的作用的体外和体内模型。包括分离和培养骨髓来源的巨噬细胞,收获肿瘤和巨噬细胞来源的条件培养基,以及研究条件培养基对C2C12肌管的影响。还描述了涉及使用胰腺癌原位模型的程序,包括在该模型中检查骨骼肌萎缩的方法。©2020 Wiley期刊有限公司。基本方案1:利用C2C12细胞系(肌管模型)建立胰腺肿瘤诱导的恶病质体外模型。支持方案1:利用实时PCR和免疫印迹技术对C2C12肌管中恶病质标志物的分子评价。基本方案2:在体内模型研究胰腺肿瘤小鼠的恶病质表型。支持方案2:评估肿瘤小鼠骨骼肌中的恶病质标志物
本文章由计算机程序翻译,如有差异,请以英文原文为准。