Name: the colon-26 carcinoma tumor-bearing mouse as a model for the study of cancer cachexia
Uploaded: 2016-11-30T14:00:00
Description: Watch this Scientific Journal Video about The Colon-26 Carcinoma Tumor-bearing Mouse as a Model for the Study of Cancer Cachexia at JoVE.com

We thank Richard Lieber and Shannon Bremner for their ImageJ macro and instructions. While at Thomas Jefferson University, this work was supported by the Pennsylvania Department of Health CURE Grant TJU No. 080-37038-AI0801. Subsequently, this study was supported by a grant to AB from the National Institutes of Health (R21CA190028), and by grants to TAZ from the National Institutes of Health (R01CA122596, R01CA194593), the IU Simon Cancer Center, the Lustgarten Foundation, the Lilly Foundation, Inc., and the IUPUI Pancreas Signature Center.

倫理文：説明すべての研究は、医学のトーマス・ジェファーソン大学とインディアナ大学の施設内動物管理使用委員会によって承認されました。 1. C26細胞成長と準備<ol><li> 、1％、10％ウシ胎児血清（FBS）、1mMピルビン酸ナトリウムを含有する（ すなわち、高グルコースのダルベッコ改変イーグル培地（DMEM）C26大腸癌細胞（オハイオ州立大学医療センター（OSUMC?...

<ol>
	<li>Tan, B., Fearon, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cachexia:+prevalence+and+impact+in+medicine.">Cachexia: prevalence and impact in medicine.</a> Curr Opin Clin Nutr Metab Care. 11, 400-407 (2008).</li><li>Aulino, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Molecular+cellular+and+physiological+characterization+of+the+cancer+cachexia-inducing+C26+colon+carcinoma+in+mouse.">Molecular cellular and physiological characterization of the cancer cachexia-inducing C26 colon carcinoma in mouse.</a> BMC cancer. 10, 363 (2010).</li><li>Bonetto, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=STAT3+activation+in+skeletal+muscle+links+muscle+wasting+and+the+acute+phase+response+in+cancer+cachexia.">STAT3 activation in skeletal muscle links muscle wasting and the acute phase response in cancer cachexia.</a> PloS One. 6, e22538 (2011).</li><li>Bonetto, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=JAK/STAT3+pathway+inhibition+blocks+skeletal+muscle+wasting+downstream+of+IL-6+and+in+experimental+cancer+cachexia.">JAK/STAT3 pathway inhibition blocks skeletal muscle wasting downstream of IL-6 and in experimental cancer cachexia.</a> Am J Physiol Endocrinol Metab. 303, E410-E421 (2012).</li><li>Bonetto, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Deacetylase+inhibitors+modulate+the+myostatin/follistatin+axis+without+improving+cachexia+in+tumor-bearing+mice.">Deacetylase inhibitors modulate the myostatin/follistatin axis without improving cachexia in tumor-bearing mice.</a> Current Cancer Drug Targets. 9, 608-616 (2009).</li><li>Acharyya, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+cachexia+is+regulated+by+selective+targeting+of+skeletal+muscle+gene+products.">Cancer cachexia is regulated by selective targeting of skeletal muscle gene products.</a> The Journal of Clinical Investigation. 114, 370-378 (2004).</li><li>Fearon, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Definition+and+classification+of+cancer+cachexia:+an+international+consensus.">Definition and classification of cancer cachexia: an international consensus.</a> The Lancet Oncology. 12, 489-495 (2011).</li><li>Holecek, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+wasting+in+animal+models+of+severe+illness.">Muscle wasting in animal models of severe illness.</a> Int J Exp Pathol. 93, 157-171 (2012).</li><li>Acharyya, S., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Dystrophin+glycoprotein+complex+dysfunction:+a+regulatory+link+between+muscular+dystrophy+and+cancer+cachexia.">Dystrophin glycoprotein complex dysfunction: a regulatory link between muscular dystrophy and cancer cachexia.</a> Cancer Cell. 8, 421-432 (2005).</li><li>Benny Klimek, ., E, M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Acute+inhibition+of+myostatin-family+proteins+preserves+skeletal+muscle+in+mouse+models+of+cancer+cachexia.">Acute inhibition of myostatin-family proteins preserves skeletal muscle in mouse models of cancer cachexia.</a> Biochemical and Biophysical Research Communications. 391, 1548-1554 (2010).</li><li>Pedroso, F. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Inflammation,+organomegaly,+and+muscle+wasting+despite+hyperphagia+in+a+mouse+model+of+burn+cachexia.">Inflammation, organomegaly, and muscle wasting despite hyperphagia in a mouse model of burn cachexia.</a> Journal of Cachexia, Sarcopenia and Muscle. 3, 199-211 (2012).</li><li>Soda, K., Kawakami, M., Kashii, A., Miyata, M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Manifestations+of+cancer+cachexia+induced+by+colon+26+adenocarcinoma+are+not+fully+ascribable+to+interleukin-6.">Manifestations of cancer cachexia induced by colon 26 adenocarcinoma are not fully ascribable to interleukin-6.</a> International journal of cancer. 62, 332-336 (1995).</li><li>Costelli, P., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=IGF-1+is+downregulated+in+experimental+cancer+cachexia.+American+journal+of+physiology.">IGF-1 is downregulated in experimental cancer cachexia. American journal of physiology.</a> Regulatory, Integrative and Comparative Physiology. 291, R674-R683 (2006).</li><li>Palus, S., Akashi, Y., von Haehling, S., Anker, S. D., Springer, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+influence+of+age+and+sex+on+disease+development+in+a+novel+animal+model+of+cardiac+cachexia.">The influence of age and sex on disease development in a novel animal model of cardiac cachexia.</a> International Journal of Cardiology. 133, 388-393 (2009).</li><li>Norman, K., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Effect+of+sexual+dimorphism+on+muscle+strength+in+cachexia.">Effect of sexual dimorphism on muscle strength in cachexia.</a> Journal of Cachexia, Sarcopenia and Muscle. 3, 111-116 (2012).</li><li>Stephens, N. A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Sexual+dimorphism+modulates+the+impact+of+cancer+cachexia+on+lower+limb+muscle+mass+and+function.">Sexual dimorphism modulates the impact of cancer cachexia on lower limb muscle mass and function.</a> Clinical Nutrition. 31, 499-505 (2012).</li><li>Cosper, P. F., Leinwand, L. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Cancer+causes+cardiac+atrophy+and+autophagy+in+a+sexually+dimorphic+manner.">Cancer causes cardiac atrophy and autophagy in a sexually dimorphic manner.</a> Cancer Research. 71, 1710-1720 (2011).</li><li>Ullman-Cullere, M. H., Foltz, C. J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Body+condition+scoring:+a+rapid+and+accurate+method+for+assessing+health+status+in+mice.">Body condition scoring: a rapid and accurate method for assessing health status in mice.</a> Laboratory Animal Science. 49, 319-323 (1999).</li><li>Bonetto, A., Andersson, D. C., Waning, D. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Assessment+of+muscle+mass+and+strength+in+mice.">Assessment of muscle mass and strength in mice.</a> Bonekey Rep. 4, 732 (2015).</li><li>Minamoto, V. B., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Increased+efficacy+and+decreased+systemic-effects+of+botulinum+toxin+A+injection+after+active+or+passive+muscle+manipulation.">Increased efficacy and decreased systemic-effects of botulinum toxin A injection after active or passive muscle manipulation.</a> Dev Med Child Neurol. 49, 907-914 (2007).</li><li>Murphy, K., Lynch, G. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Update+on+emerging+drugs+for+cancer+cachexia.">Update on emerging drugs for cancer cachexia.</a> Expert Opin Emerg Drugs. 14, 619-632 (2009).</li><li>Seto, D. N., Kandarian, S. C., Jackman, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+Key+Role+for+Leukemia+Inhibitory+Factor+in+C26+Cancer+Cachexia.">A Key Role for Leukemia Inhibitory Factor in C26 Cancer Cachexia.</a> The Journal of Biological Chemistry. 290, 19976-19986 (2015).</li><li>Judge, S. M., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Genome-wide+identification+of+FoxO-dependent+gene+networks+in+skeletal+muscle+during+C26+cancer+cachexia.">Genome-wide identification of FoxO-dependent gene networks in skeletal muscle during C26 cancer cachexia.</a> BMC Cancer. 14, 997 (2014).</li><li>Kliewer, K. L., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Adipose+tissue+lipolysis+and+energy+metabolism+in+early+cancer+cachexia+in+mice.">Adipose tissue lipolysis and energy metabolism in early cancer cachexia in mice.</a> Cancer Biol Ther. 16, 886-897 (2015).</li><li>Aversa, Z., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Changes+in+myostatin+signaling+in+non-weight-losing+cancer+patients.">Changes in myostatin signaling in non-weight-losing cancer patients.</a> Ann Surg Oncol. 19, 1350-1356 (2012).</li><li>Bonetto, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Early+changes+of+muscle+insulin-like+growth+factor-1+and+myostatin+gene+expression+in+gastric+cancer+patients.">Early changes of muscle insulin-like growth factor-1 and myostatin gene expression in gastric cancer patients.</a> Muscle Nerve. 48, 387-392 (2013).</li><li>Lazarus, D. D., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=A+new+model+of+cancer+cachexia:+contribution+of+the+ubiquitin-proteasome+pathway.">A new model of cancer cachexia: contribution of the ubiquitin-proteasome pathway.</a> The American Journal of Physiology. 277, E332-E341 (1999).</li><li>al-Majid, S., McCarthy, D. O. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Resistance+exercise+training+attenuates+wasting+of+the+extensor+digitorum+longus+muscle+in+mice+bearing+the+colon-26+adenocarcinoma.">Resistance exercise training attenuates wasting of the extensor digitorum longus muscle in mice bearing the colon-26 adenocarcinoma.</a> Biol Res Nurs. 2, 155-166 (2001).</li><li>Bonetto, A., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=JAK/STAT3+pathway+inhibition+blocks+skeletal+muscle+wasting+downstream+of+IL-6+and+in+experimental+cancer+cachexia.">JAK/STAT3 pathway inhibition blocks skeletal muscle wasting downstream of IL-6 and in experimental cancer cachexia.</a> American journal of physiology. Endocrinology and metabolism 303. 303, E410-E421 (2012).</li><li>Samuels, S. E., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Liver+protein+synthesis+stays+elevated+after+chemotherapy+in+tumour-bearing+mice.">Liver protein synthesis stays elevated after chemotherapy in tumour-bearing mice.</a> Cancer Lett. 239, 78-83 (2006).</li><li>Cornwell, E. W., Mirbod, A., Wu, C. L., Kandarian, S. C., Jackman, R. W. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=C26+cancer-induced+muscle+wasting+is+IKKbeta-dependent+and+NF-kappaB-independent.">C26 cancer-induced muscle wasting is IKKbeta-dependent and NF-kappaB-independent.</a> PloS One. 9, e87776 (2014).</li><li>Penna, F., et al. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Muscle+wasting+and+impaired+myogenesis+in+tumor+bearing+mice+are+prevented+by+ERK+inhibition.">Muscle wasting and impaired myogenesis in tumor bearing mice are prevented by ERK inhibition.</a> PloS One. 5, e13604 (2010).</li></ol>

特にその最新の段階で、結腸直腸癌は、貧しい転帰と患者の生活の質の低下の原因である悪液質の開発と関連しています。多くの研究は、癌の二次的症状の治療に焦点を当てています。しかし、この方向に多くの努力にもかかわらず、癌性悪液質21には承認された治療法はまだありません。従って、動物モデルは、所見の変換を最大にするためにできるだけ近いヒト病理に似ているこ...

筋萎縮は、癌、敗血症、肝臓、肝硬変、心臓や腎不全、慢性閉塞性肺疾患、およびAIDSなどの様々な臨床症状の重篤な合併症です。特に、筋消耗、癌1患者の少なくとも50％において明らかです。原因および/またはからの骨格筋タンパク質分解システムの過剰活性化に増加したタンパク質分解からの癌の結果で骨格筋の損失は、タンパク質合成6を減少させました 。脂肪分解は、脂肪組織の枯渇をもたらすも明らかです。臨床的には、悪液質を低減し、品質及び寿命の長さと関連し、20における死亡の原因であると推定されている-癌患者7の30％。できるだけヒトの疾患に似ている実験モデルの使用は有益であろう。最適な動物モデルは、異なる治療法との予測不可能な要因から高い再現性により、同様に制限された干渉によって特徴づけられます通常、臨床状態8に関連付けられている食事、性別、遺伝的背景。新しい方法は、癌の発生に影響を受けやすい遺伝的に改変されたマウスを使用することであるが、これまで、癌性悪液質は、癌細胞または発癌物質の注入の移植によって特徴付けられる動物モデルにおいて主に研究されてきました。 C26癌を有するマウスは、癌性悪液質2,5のよく特徴付けられたと広範囲に使用されるモデルを表す（結腸-26および腺癌と呼ばれます）。主に強化脂肪及びタンパク質異化9を介して身体および筋肉重量損失C26腫瘍結果の成長。一般に、全体重に対して10％の腫瘍重量は、骨格筋重量の20％〜25％の減少および脂肪3,10の高い枯渇と関連しています。肝腫および脾腫も、急性期反応の活性化及びプロinflaの上昇に伴って、腫瘍の成長で観察されていますmmatoryサイトカインレベル3,11。これらの中でも、同様にこのサイトカインはおそらく、悪液質12の唯一の誘導因子ではないにもかかわらず、IL-6は、C26モデルにおいて筋肉消耗を媒介する重要な役割を果たしていることが知られています。上昇したIL-6は、JAK / STAT3経路の活性化を介して筋萎縮を引き起こし、この転写因子を阻害すること3,4筋肉消耗を防ぐことができます。 C26誘発性筋萎縮時には、筋萎縮の多くの条件のように、筋肉量がない細胞死または繊維13の損失を介して、主に筋線維全体の筋肉のタンパク質含有量の減少によって失われます。 C26の悪液質では、より小さな断面積へのシフトは、解糖および酸化繊維2の両方で観察されました。これはまた、減少筋力5と一致しています。多くのグループが、世界中のがんCACのための筋萎縮の新しいメディエーターまたは臨床的に関連する薬物を発見するために、C26モデルの利点をとっていますhexia。しかし、このモデルの使用のために多くの異なる手順は、得られたデータの整合性についての懸念を高め、異なる実験条件で再現性の障壁を装った、報告されています。ここでは、標準化と再現性のあるデータが得られ、癌性悪液質の研究のためのこのモデルの典型的な使用を報告しています。

<table border="0" cellpadding="0" cellspacing="0" width="661">
	<tbody>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Cell culture Flasks</td>
			<td style="width:205px;">Falcon - Becton Dickinson</td>
			<td style="width:104px;">35-5001</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">DMEM</td>
			<td style="width:205px;">Cellgro</td>
			<td style="width:104px;">10-017-CV</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">FBS</td>
			<td style="width:205px;">Gibco</td>
			<td style="width:104px;">26140</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Streptomycin-Penicillin&#160;</td>
			<td style="width:205px;">Cellgro</td>
			<td style="width:104px;">30-002-CI</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">CD2F1 mice</td>
			<td>Harlan</td>
			<td>060</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Anesthesia apparatus</td>
			<td style="width:205px;">EZ-Anesthesia</td>
			<td style="width:104px;">EZ-7000</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">2-Methyl Butane</td>
			<td style="width:205px;">Sigma-Aldrich</td>
			<td style="width:104px;">M32631</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">OCT</td>
			<td style="width:205px;">Tissue-Tek</td>
			<td style="width:104px;">4583</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Cryostat</td>
			<td style="width:205px;">Leica</td>
			<td style="width:104px;">CM1850</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Cork disks</td>
			<td>Electron Microscopy Sciences</td>
			<td>63305</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:167px;">Superfrost plus glass slides</td>
			<td>VWR</td>
			<td>48311-703</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:167px;">Anti-Laminin Rabbit polyclonal antibody</td>
			<td>Sigma-Aldrich</td>
			<td>L9393</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:167px;">Anti-Dystrophin Mouse Monoclonal antibody</td>
			<td>Vector Laboratories</td>
			<td>VP-D508</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:167px;">Alexa Flour 594 anti-mouse IgG</td>
			<td>Life Technologies</td>
			<td>A11062</td>
			<td></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:167px;">Alexa Flour 594 anti-rabbit IgG</td>
			<td>Life Technologies</td>
			<td>A21211</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Hematoxylin</td>
			<td>Sigma-Aldrich</td>
			<td>GHS216</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Eosin</td>
			<td>Sigma-Aldrich</td>
			<td>HT110332</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;">Xylene</td>
			<td>Acros Organics</td>
			<td>422680025</td>
			<td></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Cytoseal-XYL</td>
			<td style="width:205px;">Thermo</td>
			<td style="width:104px;">8312-4</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Microscope</td>
			<td style="width:205px;">Zeiss</td>
			<td style="width:104px;">Observer.Z1&#160;</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Bamboo Tablet</td>
			<td style="width:205px;">Wacom</td>
			<td style="width:104px;">CTH-661</td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Prism 7.0 for Mac OS X</td>
			<td style="width:205px;">GraphPad Software, Inc.</td>
			<td style="width:104px;"></td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="21">
			<td height="21" style="height:21px;width:167px;">Excel for Mac 2011</td>
			<td style="width:205px;">Microsoft Corp.</td>
			<td style="width:104px;"></td>
			<td style="width:185px;"></td>
		</tr>
		<tr height="42">
			<td height="42" style="height:42px;width:167px;">Image J</td>
			<td style="width:205px;">US National Institutes of Health</td>
			<td style="width:104px;">IJ1.46</td>
			<td style="width:185px;">http://rsbweb.nih.gov/ij/download.html</td>
		</tr>
		<tr height="22">
			<td height="22" style="height:22px;width:167px;">Microtainer</td>
			<td style="width:205px;">BD</td>
			<td style="width:104px;">365873</td>
			<td style="width:185px;"></td>
		</tr>
	</tbody>
</table>

the colon-26 carcinoma tumor-bearing mouse as a model for the study of cancer cachexia

Cancer cachexia is the progressive loss of skeletal muscle mass and adipose tissue, negative nitrogen balance, anorexia, fatigue, inflammation, and activation of lipolysis and proteolysis systems. Cancer patients with cachexia benefit less from anti-neoplastic therapies and show increased mortality1. Several animal models have been established in order to investigate the molecular causes responsible for body and muscle wasting as a result of tumor growth. Here, we describe methodologies pertaining to a well-characterized model of cancer cachexia: mice bearing the C26 carcinoma2-4. Although this model is heavily used in cachexia research, different approaches make reproducibility a potential issue. The growth of the C26 tumor causes a marked and progressive loss of body and skeletal muscle mass, accompanied by reduced muscle cross-sectional area and muscle strength3-5. Adipose tissue is also lost. Wasting is coincident with elevated circulating levels of pro-inflammatory cytokines, particularly Interleukin-6 (IL-6)3, which is directly, although not entirely, responsible for C26 cachexia. It is well-accepted that a primary mechanism by which the C26 tumor induces muscle tissue depletion is the activation of skeletal muscle proteolytic systems. Thus, expression of muscle-specific ubiquitin ligases, such as atrogin-1/MAFbx and MuRF-1, represent an accepted method for the evaluation of the ongoing muscle catabolism2. Here, we present how to execute this model in a reproducible manner and how to excise several tissues and organs (the liver, spleen, and heart), as well as fat and skeletal muscles (the gastrocnemius, tibialis anterior, and quadriceps). We also provide useful protocols that describe how to perform muscle freezing, sectioning, and fiber size quantification.

（ - 5 D 4）指数関数的な細胞増殖が続く注射後8 D、 - C26腫瘍増殖動態は、最初の7のための誘導期を示しました。腫瘍塊は、最終的に（; 図 1A-B約2g）〜体重の10％に達します。最初のフェーズでは、腫瘍は触診により配置することができ、皮膚の小さな突起として表示されます。第二段階では、腫瘍は、皮膚の下の塊として観察されます。ま?...

Watch this Scientific Journal Video about The Colon-26 Carcinoma Tumor-bearing Mouse as a Model for the Study of Cancer Cachexia at JoVE.com

The Colon-26 Carcinoma Tumor-bearing Mouse as a Model for the Study of Cancer Cachexia

Mice bearing the Colon-26 (C26) carcinoma represent a classical model of cancer cachexia. Progressive muscle wasting occurs in association with tumor growth, over-expression of muscle-specific ubiquitin ligases, and reductions in muscle cross-sectional area. Fat loss is also observed. Cachexia is studied in a time-dependent manner with increasing severity of wasting.

大腸-26癌腫瘍を有するがん悪液質の研究のためのモデルとしてマウスを

Cancer cachexia is the progressive loss of skeletal muscle mass and adipose tissue, negative nitrogen balance, anorexia, fatigue, inflammation, and ...

The overall goals of this protocol are to execute the Colon 26 In Vivo model in a reproducible manor, and to properly investigate the progressive muscle wasting and weakness that occur in association with tumor growth. This model can help answer key questions in a cancer cachexia field about tumor induced muscle loss, muscle weakness, increased muscle catabolism, and hyper inflammatory cytokine levels. The main advantage of this experimental model is that change is consistent with a cachectic phenotype can be easily started in a time dependent manor as the wasting severity increases.Demonstrating these procedures will be Rafael Barreto, a technician from my lab, and Joseph Rupert, a PhD student from Teresa Zimmer's group. To implant the C 26 tumors use a sterile, one millimeter insulin syringe, equipped with a 26 gauge needle to quickly inject one times 10 to the sixth cells subcutaneously and interscapularly into the fat pads of individual, adult, CD two F one mice. Return the animals to their cage after tumor injection, with monitoring.Checking the mice, and recording their body weights daily. When the body weight loss in the tumor host, versus the control animals, reaches five, 10, or 15%compared to their initial body weights, draw one to one point five milliliters of blood by cardiac puncture, and transfer the individual blood samples into empty 18 milligram potassium EDTA tubes. Place the tubes on ice until all of the blood has been collected.Then, centrifuge the samples, collecting the plasma and serum at the end of the centrifugation. To avoid further contamination, spray 70%ethanol over the entire body. Then place the animal on a dissection bed in the supine position, and attach one hind limb to an elevated burette clamp to extend the limb vertically.Using Dumont forceps, grasp the superficial skin of the hind limb and use curved fine scissors to gently remove the skin and fascia, exposing the underlying muscle bellies of the lower limb. Identify the gastrocnemius muscle on the posterior lower limb by its origin at the lateral and medial condyles on the posterior femur, and its insertion at the calcaneus, vial the calcaneal tendon. Use the forceps to grasp the distal end of the gastrocnemius and pull the muscle belly toward it's origin.Then, use scissors to cut the muscle at its origin, as close as possible to the femur and transfer the muscle to a pre-weighed dish. Weigh the dish immediately, and transfer the muscle to a pre-labeled cryo tube. Next, identify the tibialis anterior muscle from its origin at the anterolateral surface of the tibia and its insertion at the medial cuneiform via its distal tendon.Then, grasp the foot by the digits with the index finger and thumb, and insert the tip of the Dumont forceps immediately under the superficial distal tendon of the tibialis. Move the forceps such that the blunt side can be used to detach the tibialis muscle belly from the underlying connective tissue, and use the fine curved scissors to cut the distal tendon. Then, cut the muscle at the origin, as close as possible to the tibia.When all of the muscles have been collected, immerse the bottom half of a plastic, 50 milliliter beaker containing isopentane into liquid nitrogen. When the isopentane becomes slightly viscous, with a solid white laminate lining visible inside the beaker, pour a few drops of embedding medium on a chuck. Then, pick up a muscle by the tendon, and carefully position the end of the fresh muscle on top of the embedding medium.Holding it vertically, relative to the cork. Then, keeping the muscle vertical, dip the chuck and attached muscle into the isopentane bath for seven to 15 seconds until the tissue is completely frozen. A well frozen specimen will appear chalky white.Then, place the muscle in a specimen tube for immediate storage at negative 80 degrees Celsius. To preserve the RNA and enzymatic structure and tibialis samples, it is required to freeze the muscle tissues as soon as possible after their extraction. To section the frozen muscle tissue, first set the working temperature of the Cryostat inner chamber to negative 23 degrees Celsius.Allow the negative 80 degrees Celsius specimen to acclimate to the working temperature for a few hours. Then, obtain multiple eight micron thick sections of the specimen at the mid belly region of the muscle, perpendicular to the mounting axis. Collecting the sections on room temperature glass slides as they are cut.Keep the samples inside the Cryostat chamber until all of the sections have been collected. Then, store the samples at negative 80 degrees Celsius until further analysis. C 26 tumor growth kinetics demonstrate a lag phase for the first seven to eight days after injection, followed by exponential cell growth for four to five days.With the tumor mass eventually reaching approximately 10%of the animal's body weight. Tumor bearing mice appear wasted and exhibit disheveled fur at the end of the experimental period. With 20 to 30%reduction in skeletal muscle weight.The cardiac muscle is also significantly reduced in weight. Although to a lesser extent compared to the other muscles. Interestingly, hepatomegaly and splenomagly are also generally detected in tumor hosts.While the fat mass, similar to the skeletal muscle mass, is severely depleted. Skeletal muscle weight loss is also consistent with, and proportional to, the reduction in muscle fiber size. As observed by immunofluorescent morphometric evaluation of muscle fiber cross-sectional areas.In particular, the frequency distribution analysis demonstrates a shift towards smaller sized fibers in C 26 bearing mice. Suggesting that the whole muscle undergoes atrophy in the presence of a C 26 tumor. Similar results can also be observed by HNE analysis.Although the magnitude of change in the muscle cross-sectional area associated with the cancer growth is slightly different. Proper use of a C 26 model allows for the investigation of tumor derived cachexia. The model closely resembles a human disease showing an overall reduction in mass as well as muscle fiber atrophy, inflammation, and hyper catabolism.When performing this experiment, the strain of six of the mice, the source, and number of tumor cells, and the site of implantation can be modified. Such modifications can lead to differences in the expected outcomes. Despite a few known limitations, such as the non-physiologic growth environment, the dependency on IL six action, and the necessary use of CD two F one or Valve C mice, the C 26 model has greatly contributed to understanding of the molecular mechanisms of cancer induced cachexia.After watching this video, you should have a good understanding of how to replicate the C 26 cancer cachexia model to study the progressive muscle wasting and weakness that typically occur, concurrent with tumor development. Be aware that working with the C 26 model requires familiarity with proper animal handling techniques. Best practices should always be used to execute this protocol as humanely as possible.

Research

JoVE Journal

Cancer Research

Fluorescent Orthotopic Mouse Model of Pancreatic Cancer

膵臓がんの蛍光同所性マウスモデル

Pancreatic cancer remains one of the cancers for which survival has not improved substantially in the last few decades. Only 7% of diagnosed patients will ...

がん研究

Study of Viral Vectors in a Three-dimensional Liver Model Repopulated with the Human Hepatocellular Carcinoma Cell Line HepG2

ヒト肝細胞癌細胞株HepG2細胞と三次元肝モデルに再増殖におけるウイルスベクターの研究

This protocol describes the generation of a three-dimensional (3D) ex vivo liver model and its application to the study and development of viral vector ...

A Model for Perineural Invasion in Head and Neck Squamous Cell Carcinoma

頭頸部扁平上皮癌における神経周囲の浸潤のためのモデル

Perineural invasion (PNI) is found in approximately 40% of head and neck squamous cell carcinomas (HNSCC). Despite multimodal treatment with surgery, ...

A Syngeneic Mouse Model of Metastatic Renal Cell Carcinoma for Quantitative and Longitudinal Assessment of Preclinical Therapies

定量的および前臨床治療の縦評価のための転移性腎細胞癌の同系マウスモデル

Renal cell carcinoma (RCC) affects &#62; 60,000 people in the United States annually, and ~ 30% of RCC patients have multiple metastases at the time of ...

A Syngeneic Pancreatic Cancer Mouse Model to Study the Effects of Irreversible Electroporation

不可逆エレクトロポレーションの効果を研究する同系膵癌マウス モデル

Pancreatic cancer (PC), a disease which kills approximately 40,000 patients each year in the US, has successfully evaded several therapeutic approaches ...

Yeast As a Chassis for Developing Functional Assays to Study Human P53

ヒトP53を研究する機能アッセイを開発するシャーシとしての酵母

The finding that the well-known mammalian P53 protein can act as a transcription factor (TF) in the yeast S. cerevisiae has allowed for the development of ...

Transfer of Manipulated Tumor-associated Neutrophils into Tumor-Bearing Mice to Study their Angiogenic Potential In Vivo

操作された腫瘍関連好中球を腫瘍性マウスに移植し、生体内の血管新生電位を研究する

The contribution of neutrophils to the regulation of tumorigenesis is getting increased attention. These cells are heterogeneous, and depending on the ...

Isolation of Primary Cancer-Associated Fibroblasts from a Syngeneic Murine Model of Breast Cancer for the Study of Targeted Nanoparticles

標的ナノ粒子の研究のための乳癌の同質性マウスモデルからの原発癌関連線維芽細胞の分離

Cancer-associated fibroblasts (CAFs) are key actors in the context of the tumor microenvironment. Despite being reduced in number as compared to tumor ...

A Three-Dimensional Spheroid Model to Investigate the Tumor-Stromal Interaction in Hepatocellular Carcinoma

肝細胞癌における腫瘍間質相互作用を調べる三次元スフェロイドモデル

The aggressiveness and lack of well-tolerated and widely effective treatments for advanced hepatocellular carcinoma (HCC), the predominant form of liver ...

Molecular and Immunologic Techniques in a Genetically Engineered Mouse Model of Gastrointestinal Stromal Tumor

消化管間質腫瘍の遺伝子操作マウスモデルにおける分子および免疫学的技術

Gastrointestinal stromal tumor (GIST) is the most common human sarcoma and is typically driven by a single mutation in the KIT receptor. Across tumor ...