Name: zebrafish model of neuroblastoma metastasis
Uploaded: 2021-03-14T15:00:00
Description: Watch this Scientific Journal Video about Zebrafish Model of Neuroblastoma Metastasis at JoVE.com

这项工作得到了国家癌症研究所的R01 CA240323（S.Z.）资助;美国国防部（DoD）的拨款W81XWH-17-1-0498（S.Z.）;V癌症研究基金会（S.Z.）的V学者奖和梅奥生物医学发现中心（S.Z.）的平台资助;以及梅奥诊所癌症中心和个性化医学中心（S.Z.）的支持。

所有使用斑马鱼和动物护理/维护的研究方法均按照梅奥诊所的机构指南进行。
1. PSNS中过表达的转基因构建体开发 转基因 构建体的制备和显微注射
<ol><li>为了开发 LMO1-pDONR221 进入克隆，使用PCR扩增从人细胞系获得的cDNA中人类 LMO1 的编码区域。<ol>
<li>制作25μL反应，详见：2.5μL1标准Taq反应缓冲液，0.125μLTaq DNA聚合酶，0.5μL10mM dNTPs，2μLcDNA模板，0.5?...

<ol>
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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=Microenvironment-derived+factors+driving+metastatic+plasticity+in+melanoma.">Microenvironment-derived factors driving metastatic plasticity in melanoma.</a> Nature Communications. 8, 14343 (2017).</li><li>Zhu, 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=Activated+ALK+collaborates+with+MYCN+in+neuroblastoma+pathogenesis.">Activated ALK collaborates with MYCN in neuroblastoma pathogenesis.</a> Cancer Cell. 21 (3), 362-373 (2012).</li><li>Maris, J. M., Hogarty, M. D., Bagatell, R., Cohn, S. L. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Neuroblastoma.">Neuroblastoma.</a> Lancet. 369 (9579), 2106-2120 (2007).</li><li>Park, J. 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C., 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+developmental+model+of+neuroblastoma:+differentiating+stroma-poor+tumors'+progress+along+an+extra-adrenal+chromaffin+lineage.">A developmental model of neuroblastoma: differentiating stroma-poor tumors' progress along an extra-adrenal chromaffin lineage.</a> Laboratory Investigation: A Journal of Technical Methods and Pathology. 75 (5), 659-675 (1996).</li><li>Tsubota, S., Kadomatsu, K. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Origin+and+initiation+mechanisms+of+neuroblastoma.">Origin and initiation mechanisms of neuroblastoma.</a> Cell and Tissue Research. 372 (2), 211-221 (2018).</li><li>Tolbert, V. P., Matthay, K. 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M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Recent+advances+in+neuroblastoma.">Recent advances in neuroblastoma.</a> New England Journal of Medicine. 362 (23), 2202-2211 (2010).</li><li>Zhu, 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=LMO1+Synergizes+with+MYCN+to+promote+neuroblastoma+initiation+and+metastasis.">LMO1 Synergizes with MYCN to promote neuroblastoma initiation and metastasis.</a> Cancer Cell. 32, 310-323 (2017).</li><li>Patton, E. E., Zon, L. I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+art+and+design+of+genetic+screens:+zebrafish.">The art and design of genetic screens: zebrafish.</a> Nature Reviews Genetics. 2 (12), 956-966 (2001).</li><li>Lieschke, G. J., Currie, P. 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W., 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=MYC+drives+a+subset+of+high-risk+pediatric+neuroblastomas+and+is+activated+through+mechanisms+including+enhancer+hijacking+and+focal+enhancer+amplification.">MYC drives a subset of high-risk pediatric neuroblastomas and is activated through mechanisms including enhancer hijacking and focal enhancer amplification.</a> Cancer Discovery. 8 (3), 320-335 (2018).</li><li>Koach, J., 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=Drugging+MYCN+oncogenic+signaling+through+the+MYCN-PA2G4+binding+interface.">Drugging MYCN oncogenic signaling through the MYCN-PA2G4 binding interface.</a> Cancer Research. 79 (21), 5652-5667 (2019).</li><li>Kimmel, C. B., Ballard, W. W., Kimmel, S. R., Ullmann, B., Schilling, T. 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I. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Stem+cell+safe+harbor:+The+hematopoietic+stem+cell+niche+in+zebrafish.">Stem cell safe harbor: The hematopoietic stem cell niche in zebrafish.</a> Blood Advances. 2 (21), 3063-3069 (2018).</li><li>Menke, A. L., Spitsbergen, J. M., Wolterbeek, A. P., Woutersen, R. A. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Normal+anatomy+and+histology+of+the+adult+zebrafish.">Normal anatomy and histology of the adult zebrafish.</a> Toxicologic Pathology. 39 (5), 759-775 (2011).</li><li>Renshaw, S. A., Trede, N. 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在过去的几十年里，斑马鱼一直被普遍用于研究，特别是在癌症研究中，原因显而易见，例如其易于维护，强大的繁殖以及 体内成像的 明显优势1，28。由于斑马鱼模型的外部受精和发育，它们可以很容易地在胚胎上纵，这与哺乳动物模式生物（如大鼠和小鼠）相辅相成，用于大规模的遗传研究 1 ， 2<s...

斑马鱼已被广泛使用并应用于多个研究领域，特别是在癌症方面。该模型提供了许多优点 - 例如其强大的繁殖，经济高效的维护以及肿瘤生长和转移的多功能可视化 - 所有这些都使斑马鱼成为研究和研究肿瘤发生和转移的细胞和分子基础的强大工具。用于大规模基因组图谱、转基因、基因过表达或敲除、细胞移植和化学筛选的新技术极大地增强了斑马鱼模型的力量1。在过去的几年中，已经开发了许多斑马鱼系来研究各种人类癌症的肿瘤发生和转移，包括但不限于白血病，黑色素瘤，横纹肌肉瘤和肝细胞癌2，3，4，5。此外，第一个斑马鱼神经母细胞瘤（NB）模型是通过在多巴胺-β-羟化酶（dβh）启动子的控制下在外周交感神经系统（PSNS）中过度表达MYCN（一种癌基因）而产生的。通过该模型，进一步证明激活的ALK可以与MYCN协同作用，以加速肿瘤发作并增加体内肿瘤外显率6。
NB源自神经嵴细胞的交感肾上腺谱系，是儿童中高度转移的癌症7。它占儿科癌症相关死亡的10%8。NB在诊断时广泛转移，临床上可表现为主要起源于交感神经节链和PSNS9，10的肾上腺髓链的肿瘤。 MYCN 扩增通常与 NB 患者的不良结局相关11，12。此外， LMO1 已被确定为高风险病例中的关键NB易感基因13，14。研究发现，在斑马鱼模型的PSNS中 ，MYCN 和 LMO1 的转基因共表达不仅促进了NB的早期发作，而且还诱导了与高危NB13患者常见的相似部位的组织和器官的广泛转移。最近，在较新的NB斑马鱼模型中也观察到了NB的另一种转移表型，其中编码RNA结合蛋白的 MYCN 和 Lin28B 在 dβh 启动子的控制下都过度表达16。
斑马鱼的稳定转基因方法通常用于研究目的基因的过表达是否有助于正常发育和疾病发病机制14，15。该技术已成功用于证明多个基因和途径对NB肿瘤发生的重要性6，16，17，18，19，20。本文将介绍如何在PSNS中产生过表达MYCN和LMO1的转基因鱼系，以及如何证明这两种癌基因的合作加速了NB肿瘤发生和转移的发生13。首先，通过将dβh-EGFP-MYCN构建体注射到野生型（WT）AB胚胎的单细胞阶段，开发了在dβh-EGFP-MYCN（指定为MYCN系）控制下过表达EGFP-MYCN的转基因系，如前所述6，17。通过将两个DNA构建体dβh-LMO1和dβh-mCherry在单细胞阶段13中共注射到WT胚胎中，开发了一种在PSNS（指定的LMO1系）中过表达LMO1的单独转基因系。先前已经证明，共注射的双DNA构建体可以共同整合到鱼类基因组中;因此，LMO1和mCherry在转基因动物的PSNS细胞中共表达。一旦注射的F0胚胎达到性成熟，它们就会与WT鱼杂交，以鉴定具有转基因整合的阳性鱼。简而言之，F1后代首先通过荧光显微镜筛选PSNS细胞中的mCherry表达。通过基因组PCR和测序进一步证实了MCherry阳性鱼中LMO1的种系整合。在成功鉴定各转基因品系后，将杂合MYCN和LMO1转基因鱼的后代杂交，生成表达MYCN和LMO1的复合鱼系（指定MYCN;改性活生物体1系）。含肿瘤的MYCN;通过荧光显微镜每两周监测一次LMO1鱼，以寻找远离原发部位的区域，即腺间区域（IRG，相当于人类肾上腺的斑马鱼）转移性肿瘤的证据13。确认MYCN中肿瘤的转移;应用改性活生物体1鱼组织学和免疫组化分析。

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		<tr>
			<td>3,3&#8217;-Diaminobenzidine (DAB) Vector Kit</td>
			<td>Vector</td>
			<td>SK-4100</td>
			<td></td>
		</tr>
		<tr>
			<td>Acetic Acid</td>
			<td>Fisher Scientific / Acros Organic</td>
			<td>64-19-7</td>
			<td></td>
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		<tr>
			<td>Agarose GP2</td>
			<td>Midwest Scientific</td>
			<td>009012-36-6</td>
			<td></td>
		</tr>
		<tr>
			<td>Anti-Tyrosine Hydroxylase (TH) Antibody</td>
			<td>Pel-Freez</td>
			<td>P40101</td>
			<td></td>
		</tr>
		<tr>
			<td>Avidin/Biotin Blocking Kit</td>
			<td>Vector</td>
			<td>SP-2001</td>
			<td></td>
		</tr>
		<tr>
			<td>BOND Intense R Detection</td>
			<td>Leica Biosystems</td>
			<td>DS9263</td>
			<td></td>
		</tr>
		<tr>
			<td>BOND primary antibody diluent</td>
			<td>Leica Biosystems Newcastle, Ltd.</td>
			<td>AR9352</td>
			<td></td>
		</tr>
		<tr>
			<td>BOND-MAX IHC instrument</td>
			<td>Leica Biosystems Newcastle, Ltd.</td>
			<td>N/A</td>
			<td>fully automated IHC staining system</td>
		</tr>
		<tr>
			<td>CH211-270H11 BAC clone</td>
			<td>BACPAC resources center (BRFC)</td>
			<td>N/A</td>
			<td></td>
		</tr>
		<tr>
			<td>Compound microscope equipped with DP71 camera</td>
			<td>Olympus</td>
			<td>AX70</td>
			<td></td>
		</tr>
		<tr>
			<td>Cytoseal XYL (xylene based mounting medium)</td>
			<td>Richard-Allan Scientific</td>
			<td>8312-4</td>
			<td></td>
		</tr>
		<tr>
			<td>Eosin</td>
			<td>Leica</td>
			<td>3801601</td>
			<td>ready-to-use (no preparation needed)</td>
		</tr>
		<tr>
			<td>Ethanol</td>
			<td>Carolina</td>
			<td>86-1263</td>
			<td></td>
		</tr>
		<tr>
			<td>Expand Long Template PCR System</td>
			<td>Roche Applied Science, IN</td>
			<td>11681834001</td>
			<td></td>
		</tr>
		<tr>
			<td>Gateway BP Clonase II enzyme mix</td>
			<td>Invitrogen, CA</td>
			<td>11789-020</td>
			<td></td>
		</tr>
		<tr>
			<td>Gateway LR Clonase II enzyme mix</td>
			<td>Invitrogen, CA</td>
			<td>11791-100</td>
			<td></td>
		</tr>
		<tr>
			<td>Goat anti-Rb secondary antibody (Biotinylated)</td>
			<td>Dako</td>
			<td>E0432</td>
			<td></td>
		</tr>
		<tr>
			<td>Hematoxylin Solution, Harris Modified</td>
			<td>Sigma Aldrich Chemical Company Inc. / SAFC</td>
			<td>HHS-32-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>HRP Avidin D</td>
			<td>Vector</td>
			<td>A-2004</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrochloric Acid</td>
			<td>Aqua Solutions</td>
			<td>4360-1L</td>
			<td></td>
		</tr>
		<tr>
			<td>Hydrogen Peroxide, 3%</td>
			<td>Fisher Scientific</td>
			<td>H324-500</td>
			<td></td>
		</tr>
		<tr>
			<td>I-SceI enzyme</td>
			<td>New England Biolabs, MA</td>
			<td>R0694L</td>
			<td></td>
		</tr>
		<tr>
			<td>Kanamycin sulfate</td>
			<td>Teknova, Inc.</td>
			<td>K2150</td>
			<td></td>
		</tr>
		<tr>
			<td>Kimberly-Clark Professional Kimtech Science Kimwipes</td>
			<td>Fisher Scientific</td>
			<td>34133</td>
			<td></td>
		</tr>
		<tr>
			<td>Lithium Carbonate</td>
			<td>Sigma Aldrich Chemical Company Inc. / SAFC</td>
			<td>554-13-2</td>
			<td></td>
		</tr>
		<tr>
			<td>Microtome for sectioning</td>
			<td>Leica Biosystems</td>
			<td>RM2255</td>
			<td></td>
		</tr>
		<tr>
			<td>One Shot TOP10 Chemically Competent&#160;E. coli</td>
			<td>Invitrogen</td>
			<td>C404006</td>
			<td></td>
		</tr>
		<tr>
			<td>p3E-polyA</td>
			<td>&#160;Dr. Chi-Bin Chien, Univ. of Utah</td>
			<td>N/A</td>
			<td>a generous gift 
			(Please refer to webpage http://tol2kit.genetics.utah.edu/index.php/Main_Page to obtain material, which is freely distrubted as described.)</td>
		</tr>
		<tr>
			<td>Parafin wax</td>
			<td>Surgipath Paraplast</td>
			<td>39603002</td>
			<td>Parrafin to parafin</td>
		</tr>
		<tr>
			<td>Paraformaldehyde</td>
			<td>Alfa Aesar</td>
			<td>A11313</td>
			<td></td>
		</tr>
		<tr>
			<td>pDEST vector (modified destination vector containing I-SceI recognition sites)</td>
			<td>Dr. C. Grabher, Karlsruhe Institute of Technology, Karlsruhe, Germany</td>
			<td>N/A</td>
			<td>a generous gift</td>
		</tr>
		<tr>
			<td>pDONR 221 gateway donor vector</td>
			<td>Thermo Fisher Scientific</td>
			<td>12536-017</td>
			<td></td>
		</tr>
		<tr>
			<td>pDONRP4-P1R donor vector</td>
			<td>&#160;Dr. Chi-Bin Chien, Univ. of Utah</td>
			<td>N/A</td>
			<td>a generous gift</td>
		</tr>
		<tr>
			<td>Phenol red, 0.5%</td>
			<td>Sigma Aldrich</td>
			<td>&#160;P0290</td>
			<td></td>
		</tr>
		<tr>
			<td>Phosphate Buffered Saline (PBS), 10X</td>
			<td>BioRad</td>
			<td>1610780</td>
			<td></td>
		</tr>
		<tr>
			<td>Picrosirrius red stain kit</td>
			<td>Polysciences</td>
			<td>24901-250</td>
			<td></td>
		</tr>
		<tr>
			<td>pME-mCherry</td>
			<td>Addgene</td>
			<td>26028</td>
			<td>(DBH construct)</td>
		</tr>
		<tr>
			<td>Proteinase K, recombinant, PCR Grade</td>
			<td>Roche</td>
			<td>21712520</td>
			<td></td>
		</tr>
		<tr>
			<td>QIAprep Spin MiniPrep Kit</td>
			<td>Qiagen</td>
			<td>27104</td>
			<td></td>
		</tr>
		<tr>
			<td>RDO Rapid Decalcifier</td>
			<td>Apex Enginerring</td>
			<td>RDO04</td>
			<td></td>
		</tr>
		<tr>
			<td>Sodium Azide (NaN3)</td>
			<td>Sigma Aldrich</td>
			<td>26628-22-8</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereo fluorescence microscope</td>
			<td>Leica</td>
			<td>MZ10F</td>
			<td></td>
		</tr>
		<tr>
			<td>Stereoscopic fluorescence microscope equipped with a digital sight DS-U1 camera for imaging</td>
			<td>Nikon</td>
			<td>SMZ-1500</td>
			<td></td>
		</tr>
		<tr>
			<td>Taq DNA Polymerase</td>
			<td>New England Biolabs, MA</td>
			<td>M0273L</td>
			<td></td>
		</tr>
		<tr>
			<td>Tissue-Tek VIP&#174;&#160;6 AI Vacuum Infiltration Processor</td>
			<td>Sakura</td>
			<td>N/A</td>
			<td>Model #: VIP-6-A1</td>
		</tr>
		<tr>
			<td>Tricaine-S</td>
			<td>Western Chemical Incorporated</td>
			<td>20513</td>
			<td></td>
		</tr>
		<tr>
			<td>Xylene</td>
			<td>Thermo Fisher Scientific</td>
			<td>X3P1GAL</td>
			<td></td>
		</tr>
	</tbody>
</table>

zebrafish model of neuroblastoma metastasis

斑马鱼已成为研究人类疾病，特别是癌症的重要动物模型。除了应用于斑马鱼建模的强大转基因和基因组编辑技术外，易于维护，高产量生产力和强大的实时成像功能使斑马鱼成为研究转移以及体内该过程背后的细胞和分子基础的有价值的模型系统。第一个斑马鱼神经母细胞瘤（NB）转移模型是通过在多巴胺-β-羟化酶（dβh）启动子的控制下过度表达两个癌基因MYCN和LMO1而开发的。共过表达的MYCN和LMO1导致神经母细胞发生率降低和外显率增加，以及肿瘤细胞的加速远处转移。这种新模型可靠地重申了人类转移性NB的许多关键特征，包括临床相关和转移相关遗传改变的参与;体内转移的自然和自发发展;和转移的保守部位。因此，斑马鱼模型具有剖析体内肿瘤转移复杂过程的独特优势。

为了确定LMO1是否与MYCN协同作用以影响NB发病机制，将驱动在dβh启动子控制的PSNS细胞中表达LMO1（dβh：LMO1和dβh：mCherry）或MYCN（dβh：EGFP-MYCN）表达的转基因构建体注射到斑马鱼胚胎中13。如图1A所示，在开发稳定的转基因品系并验证其基因型后，杂合MYCN和LMO1鱼进行了杂交。他们的后代分别...

Watch this Scientific Journal Video about Zebrafish Model of Neuroblastoma Metastasis at JoVE.com

Zebrafish Model of Neuroblastoma Metastasis

本文介绍了实时开发、表征和跟踪斑马鱼神经母细胞瘤模型肿瘤转移的方法，特别是在 MYCN 和 LMO1过表达的转基因斑马鱼品系中，该谱系自发地发生转移。

斑马鱼神经母细胞瘤转移模型

Zebrafish has emerged as an important animal model to study human diseases, especially cancer. Along with the robust transgenic and genome editing ...

这些方法用于开发第一个斑马鱼神经母细胞瘤转移模型。并证明该模型可以忠实地概括在神经母细胞瘤高风险患者中看到的许多转移特征。这种转移性斑马鱼模型的主要优点是肿瘤扩散的实时体内成像，以更好地了解何时以及如何发生转移。这种技术并不特定于神经母细胞瘤。它可以应用于各种类型癌症的斑马鱼模型，只要可以识别和跟踪肿瘤细胞，从而产生一系列可能性。首先，通过杂交MYCN和LMO1转基因品系，开发一个杂合转基因鱼系，过度表达MYCN和LMO1。在受精后一天，使用立体荧光显微镜对外杂交的后代进行分类，以进行EGFP表达，其表现为EGFP阳性点。分拣后，将筛选的胚胎分离到单独的培养皿中，并将培养皿标记为MYCN阳性或MYCN阴性。在受精后三到四天对两组的胚胎进行可视化和分类，以获得LMO1表达。在上颈神经节和非PSNS多巴胺能神经元细胞中寻找红色荧光斑点，特别是在后脑的长叶髓质中。在胚胎受精后五天进行筛查。然后将分类的鱼分离成四组不同的基因型，并标记为仅MYCN，仅LMO1，MYCN和LMO1以及野生类型。根据标准协议在相同的条件下饲养它们。用立体荧光显微镜观察肿瘤，用两侧的金属刮刀轻轻翻转鱼来观察肿瘤。在鉴定出可能的携带肿瘤的鱼后，将它们分离到单独的水箱中，并带有适当的标签，包括出生日期，肿瘤筛查日期和基因型。在受精后六周，重复先前的步骤以筛查含肿瘤鱼和非肿瘤鱼，以确认先前筛查过的鱼是否存在肿瘤或识别新的可能的含瘤鱼。寻找荧光阳性肿块和确诊的含瘤鱼的持续或增加的大小。在识别出含肿瘤的鱼后，每两周监测一次它们以寻找肿瘤细胞迁移的证据，其表现为远离肿瘤发生原发部位的微小EGFP或mCherry阳性肿瘤肿块。根据需要将这些鱼隔离到单独的水箱中，并适当标记以指示可能的转移。在解释LMO1鱼中的杂合性MYCN并对其后代进行分类后，EGFP MYCN表达在受精后一天在非PSNS多巴胺能神经元细胞中显着。相比之下，LMO1表达在PSNS细胞和非PSNS多巴胺能神经元细胞中都很突出。在具有MYCN和LMO1过表达的复合转基因鱼中，远离原发肿瘤部位的器官组织中检测到荧光阳性肿瘤肿块，但在仅表达MYCN的转基因鱼中未检测到。手术部分的H&E染色表明，原发性肿瘤起源于腺间区域，斑马鱼相当于人类肾上腺，这是神经母细胞瘤患者最常见的原发性疾病部位。在多个区域检测到远离腺间的肿瘤肿块，包括肾脏的远端部分，眼眶，鳃，脾脏和心脏心房的内壁。确认了原发肿瘤和所有转移部位的肿瘤细胞的PSNS神经母细胞谱系。与单独表达MYCN的肿瘤相比，在MYCN LMO1肿瘤中发现PSR染色的胶原纤维的显着扩增量和增加的厚度。使用该程序后，可以应用药物筛选和测试新的癌症疗法，这可能导致开发替代药物来预防和治疗转移性癌症。

Research

JoVE Journal

Cancer Research

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

Murine Experimental Model of Original Tumor Development and Peritoneal Metastasis via Orthotopic Inoculation with Ovarian Carcinoma Cells

原发肿瘤发展小鼠实验模型和腹膜转移<em&gt;通过</em&gt;原位接种卵巢癌细胞

Epithelial ovarian carcinoma (EOC) is associated with a poor prognosis because it shows peritoneal dissemination. To improve the prognosis, it is ...

科研

癌症研究

In Vivo Model for Testing Effect of Hypoxia on Tumor Metastasis

In Vivo Model for Testing Effect of Hypoxia on Tumor Metastasis

体内模型对缺氧对肿瘤转移的影响测试

Hypoxia has been implicated in the metastasis of Ewing sarcoma (ES) by clinical observations and in vitro data, yet direct evidence for its pro-metastatic ...

Advanced Animal Model of Colorectal Metastasis in Liver: Imaging Techniques and Properties of Metastatic Clones

肝大肠癌转移的高级动物模型：成像技术和转移性克隆性质

Patients with a limited number of hepatic metastases and slow rates of progression can be successfully treated with local treatment approaches1,2. ...

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

Surgical Procedures and Methodology for a Preclinical Murine Model of De Novo Mammary Cancer Metastasis

临床前小鼠模型的外科手术和方法<em&gt; De Novo</em&gt;乳腺癌转移

A rate-limiting aspect of transgenic mouse models of mammary adenocarcinoma is that primary tumor burden in mammary tissue typically defines study ...

A Portal Vein Injection Model to Study Liver Metastasis of Breast Cancer

门静脉注射模型来研究肝转移乳腺癌

Breast cancer is the leading cause of cancer-related mortality in women worldwide. Liver metastasis is involved in upwards of 30% of cases with breast ...

Acellular and Cellular Lung Model to Study Tumor Metastasis

脱细胞和细胞肺模型研究肿瘤转移

It is difficult to isolate tumor cells at different points of tumor progression. We created an ex vivo lung model that can show the interaction of tumor ...

Micromanipulation of Circulating Tumor Cells for Downstream Molecular Analysis and Metastatic Potential Assessment

循环肿瘤细胞的微操作, 用于下游分子分析和转移电位评估

Blood-borne metastasis accounts for&#160;most cancer-related deaths and involves circulating tumor cells (CTCs) that are successful in establishing new ...

Using Computer-based Image Analysis to Improve Quantification of Lung Metastasis in the 4T1 Breast Cancer Model

利用计算机图像分析提高4T1乳腺癌模型中肺转移的定量

Breast cancer is a devastating malignancy, accounting for 40,000 female deaths and 30% of new female cancer diagnoses in the United States in 2019 alone. ...

A Syngeneic Orthotopic Osteosarcoma Sprague Dawley Rat Model with Amputation to Control Metastasis Rate

同源原位骨肉瘤Sprague Dawley大鼠模型，截肢控制转移率

The most recent advance in the treatment of osteosarcoma (OS) occurred in the 1980s when multi-agent chemotherapy was shown to improve overall survival ...