Name: non-invasive pet/mr imaging in an orthotopic mouse model of hepatocellular carcinoma
Uploaded: 2022-08-31T13:40:00
Description: Watch this Scientific Journal Video about Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma at JoVE.com

我们感谢香港抗癌信托基金、香港研究资助局合作研究基金（CRF C7018-14E）对小动物影像实验的支持。我们亦感谢香港大学分子成像及医学回旋加速器中心（MIMCC）提供[18 F]FMISO及[18F]FDG的支持。

所有动物研究均按照香港大学比较医学研究中心（CCMR）的活体动物教学和研究委员会（CULATR）进行，该委员会是国际实验动物护理评估和认证协会（AAALAC）认可的计划。研究中使用的动物是6-8周龄的雌性BALB / cAnN-nu（裸体）小鼠，体重为20g±2g。食物和水是 随意提供的。
1.皮下注射人肝细胞癌细胞系
注意：MHCC97是一种人HCC细胞系，用于?...

<ol>
	<li>Sung, H., 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=Global+cancer+statistics+2020:+GLOBOCAN+estimates+of+incidence+and+mortality+worldwide+for+36+cancers+in+185+countries.">Global cancer statistics 2020: GLOBOCAN estimates of incidence and mortality worldwide for 36 cancers in 185 countries.</a> CA: A Cancer Journal for Clinicians. 71 (3), 209-249 (2021).</li><li>Chen, C., Lou, T. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypoxia+inducible+factors+in+hepatocellular+carcinoma.">Hypoxia inducible factors in hepatocellular carcinoma.</a> Oncotarget. 8 (28), 46691-46703 (2017).</li><li>Lu, R. -. 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=Positron-emission+tomography+for+hepatocellular+carcinoma:+Current+status+and+future+prospects.">Positron-emission tomography for hepatocellular carcinoma: Current status and future prospects.</a> World Journal of Gastroenterology. 25 (32), 4682-4695 (2019).</li><li>Larsson, 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=Adding+11C-acetate+to+18F-FDG+at+PET+examination+has+an+incremental+value+in+the+diagnosis+of+hepatocellular+carcinoma.">Adding 11C-acetate to 18F-FDG at PET examination has an incremental value in the diagnosis of hepatocellular carcinoma.</a> Molecular Imaging and Radionuclide Therapy. 21 (1), 6-12 (2012).</li><li>Huo, 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=Kinetic+analysis+of+dynamic+11C-acetate+PET/CT+imaging+as+a+potential+method+for+differentiation+of+hepatocellular+carcinoma+and+benign+liver+lesions.">Kinetic analysis of dynamic 11C-acetate PET/CT imaging as a potential method for differentiation of hepatocellular carcinoma and benign liver lesions.</a> Theranostics. 5 (4), 371-377 (2015).</li><li>Lopci, 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=PET+radiopharmaceuticals+for+imaging+of+tumor+hypoxia:+A+review+of+the+evidence.">PET radiopharmaceuticals for imaging of tumor hypoxia: A review of the evidence.</a> American Journal of Nuclear Medicine and Molecular Imaging. 4 (4), 365-384 (2014).</li><li>Mao, X., 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=Mechanisms+through+which+hypoxia-induced+caveolin-1+drives+tumorigenesis+and+metastasis+in+hepatocellular+carcinoma.">Mechanisms through which hypoxia-induced caveolin-1 drives tumorigenesis and metastasis in hepatocellular carcinoma.</a> Cancer Research. 76 (24), 7242-7253 (2016).</li><li>Kung-Chun Chiu, 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=Hypoxia+regulates+the+mitochondrial+activity+of+hepatocellular+carcinoma+cells+through+HIF/HEY1/PINK1+pathway.">Hypoxia regulates the mitochondrial activity of hepatocellular carcinoma cells through HIF/HEY1/PINK1 pathway.</a> Cell Death &amp; Disease. 10 (12), 934 (2019).</li><li>Li, Y., 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=Establishment+of+cell+clones+with+different+metastatic+potential+from+the+metastatic+hepatocellular+carcinoma+cell+line+MHCC97.">Establishment of cell clones with different metastatic potential from the metastatic hepatocellular carcinoma cell line MHCC97.</a> World Journal of Gastroenterology. 7 (5), 630-636 (2001).</li><li>Faustino-Rocha, 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=Estimation+of+rat+mammary+tumor+volume+using+caliper+and+ultrasonography+measurements.">Estimation of rat mammary tumor volume using caliper and ultrasonography measurements.</a> Lab Animal. 42 (6), 217-224 (2013).</li><li>Liu, Q., Tan, K. V., Chang, H. C., Khong, P. L., Hui, X. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Visualization+and+quantification+of+brown+and+beige+adipose+tissues+in+mice+using+[18F]+FDG+micro-PET/MR+imaging.">Visualization and quantification of brown and beige adipose tissues in mice using [18F] FDG micro-PET/MR imaging.</a> Journal of Visualized Experiments: JoVE. (173), e62460 (2021).</li><li>Lin, W. -. H., 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=Hypoxia-activated+cytotoxic+agent+tirapazamine+enhances+hepatic+artery+ligation-induced+killing+of+liver+tumor+in+HBx+transgenic+mice.">Hypoxia-activated cytotoxic agent tirapazamine enhances hepatic artery ligation-induced killing of liver tumor in HBx transgenic mice.</a> Proceedings of the National Academy of Sciences. 113 (42), 11937-11942 (2016).</li><li>Wong, T. 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=CRAF+methylation+by+PRMT6+regulates+aerobic+glycolysis-driven+hepatocarcinogenesis+via+ERK-dependent+PKM2+nuclear+relocalization+and+activation.">CRAF methylation by PRMT6 regulates aerobic glycolysis-driven hepatocarcinogenesis via ERK-dependent PKM2 nuclear relocalization and activation.</a> Hepatology. 71 (4), 1279-1296 (2020).</li><li>Yang, X., 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=Development+of+cisplatin-loaded+hydrogels+for+trans-portal+vein+chemoembolization+in+an+orthotopic+liver+cancer+mouse+model.">Development of cisplatin-loaded hydrogels for trans-portal vein chemoembolization in an orthotopic liver cancer mouse model.</a> Drug Delivery. 28 (1), 520-529 (2021).</li><li>Shi, 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=Longitudinal+evaluation+of+five+nasopharyngeal+carcinoma+animal+models+on+the+microPET/MR+platform.">Longitudinal evaluation of five nasopharyngeal carcinoma animal models on the microPET/MR platform.</a> European Journal of Nuclear Medicine and Molecular Imaging. 49 (5), 1497-1507 (2021).</li><li>Kilian, 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=Imaging+of+hypoxia+in+small+animals+with+F+fluoromisonidasole.">Imaging of hypoxia in small animals with F fluoromisonidasole.</a> Nukleonika. 61 (2), 219-223 (2016).</li><li>Kawamura, 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=Evaluation+of+optimal+post-injection+timing+of+hypoxic+imaging+with+18F-Fluoromisonidazole-PET/CT.">Evaluation of optimal post-injection timing of hypoxic imaging with 18F-Fluoromisonidazole-PET/CT.</a> Molecular Imaging and Biology. 23 (4), 597-603 (2021).</li></ol>

在这项研究中，我们描述了使用皮下肿瘤对肝原位HCC异种移植物进行HAL的程序，以及使用[18 F]FMISO和[18F]FDG PET / MR对原位异种移植物中肿瘤缺氧进行无创监测的方法。我们的兴趣在于各种癌症和疾病模型的代谢成像，用于早期诊断和治疗反应评估11，13，14，15。迄今为止，HAL HCC异种移?...

肝细胞癌 （HCC） 是全球第六大确诊癌症和第三大常见癌症死因，2020 年新增病例超过 900，000 例，死亡800，000 例 1.主要危险因素是肝硬化，这是病毒感染（乙型和丙型肝炎病毒）、酗酒、糖尿病和非酒精性脂肪性肝炎2 的结果。肝细胞癌的管理相当复杂，有几种治疗选择，包括手术切除、热消融或化学消融、移植、经动脉化疗栓塞、放疗和化疗，具体取决于疾病分期2，3。肝细胞癌是一种化疗难治性肿瘤，在根治性治疗后，多达 70% 的患者会复发2。
尽管肿瘤异质性很高，但肝细胞癌与两种常见结局有关：（i）肝细胞癌非常缺氧，（ii）肿瘤缺氧与更大的肿瘤侵袭性和治疗失败有关。HCC细胞不受控制的增殖导致血管形成前的高耗氧率，从而产生缺氧微环境。低肿瘤内氧水平会触发一系列影响肿瘤侵袭性和治疗反应的生物反应。缺氧诱导因子（HIF）通常被认为是对缺氧反应的基本转录调节因子2，3。因此，检测缺氧的能力对于可视化肿瘤组织和识别需要侵入性手术的难以接近的部位至关重要。它还有助于更好地了解导致肿瘤侵袭性的分子变化并改善患者的治疗结果。
使用正电子发射断层扫描（PET）的分子成像通常用于许多癌症（包括HCC）的诊断和分期。特别是，联合使用涉及 [18F] 氟脱氧葡萄糖 （[18F]FDG） 和 [11C] 乙酸盐的双示踪 PET 成像可显著提高肝细胞癌诊断的总体敏感性4，5。另一方面，缺氧成像可以通过使用常用的缺氧标记物[18 F]氟米索硝唑（[18F]FMISO）来实现。在临床实践中，缺氧的无创评估对于区分各种类型的肿瘤和放射治疗计划的区域非常重要6。
临床前成像已成为对不同疾病的小鼠模型进行无创和纵向评估不可或缺的工具。稳健且高度可重复的HCC模型代表了人类HCC病理生理学的临床前和转化研究以及新疗法评估的重要平台。结合PET成像，可以阐明体内行为，以在任何给定时间点提供分子水平的重要见解。在这里，我们描述了用于生成肝动脉结扎（HAL）原位HCC异种移植物的方案，并使用[18 F]FMISO和[18F]FDG PET / MR分析其体内肿瘤代谢。HAL的掺入使得转基因或化学诱导的HCC小鼠异种移植物成为研究体内肿瘤缺氧的合适模型，因为HAL可以有效阻断动脉血液供应以诱导肿瘤内缺氧7，8。此外，与使用匹莫硝唑的离体免疫组织化学染色不同，缺氧引起的肿瘤代谢变化可以很容易地使用PET成像进行无创可视化和准确定量，从而能够纵向评估治疗反应或测量耐药性的出现3，7，8.我们这里显示的方法允许创建一个强大的缺氧HCC模型，以及使用PET / MR成像对肿瘤缺氧进行无创监测，以研究体内HCC生物学。

<table border="1" fo:keep-together.within-page="1" fo:keep-with-next.within-page="always">
	<tbody>
		<tr>
			<td>0.9% sterile saline</td>
			<td>BBraun</td>
			<td>N/A</td>
			<td>0.9% sodium chloride intravenous infusion, 500 mL</td>
		</tr>
		<tr>
			<td>10# Scalpel blade</td>
			<td>RWD Life Science Co.,ltd</td>
			<td>S31010-01</td>
			<td>Animal surgery tool</td>
		</tr>
		<tr>
			<td>10% povidone-iodine solution</td>
			<td>Banitore</td>
			<td>6.425.678</td>
			<td>For disinfection</td>
		</tr>
		<tr>
			<td>25G needle with a 1 mL syringe</td>
			<td>BD PrecisionGlide</td>
			<td>N/A</td>
			<td>1 mL syringe with 25G needle for cell suspensions injections</td>
		</tr>
		<tr>
			<td>5 mL syringe</td>
			<td>Terumo</td>
			<td>SS05L</td>
			<td>5 mL syringe Luer Lock</td>
		</tr>
		<tr>
			<td>70% Ethanol</td>
			<td>Merck</td>
			<td>1.07017</td>
			<td>For disinfection</td>
		</tr>
		<tr>
			<td>Automated Cell Counter</td>
			<td>Invitrogen</td>
			<td>AMQAF2000</td>
			<td>For automated cell counting</td>
		</tr>
		<tr>
			<td>Buprenorphine</td>
			<td>HealthDirect</td>
			<td>N/A</td>
			<td>Subcutaneous injection (0.05-0.2 mg/kg/12 hours) for analgesic after surgery</td>
		</tr>
		<tr>
			<td>Cell Culture Dish (60 mm diameter)</td>
			<td>Thermo Scientific</td>
			<td>150462</td>
			<td>For tumor tissue processing</td>
		</tr>
		<tr>
			<td>Centrifuge</td>
			<td>Sigma</td>
			<td>3-16KL, fixed-angle rotor 12311</td>
			<td>For cell suspensions collection</td>
		</tr>
		<tr>
			<td>Centrifuge Conical Tube</td>
			<td>Eppendorf</td>
			<td>EP0030122151</td>
			<td>For cell suspensions collection</td>
		</tr>
		<tr>
			<td>Culture media (Dulbecco&#8217;s modified Eagle&#8217;s medium)</td>
			<td>Gibco</td>
			<td>10566024</td>
			<td>high glucose, GlutaMAX&#8482; Supplement</td>
		</tr>
		<tr>
			<td>Digital Caliper</td>
			<td>RS PRO</td>
			<td>841-2518</td>
			<td>For subcutaneous tumor size measurement</td>
		</tr>
		<tr>
			<td>Direct heat CO2 incubator</td>
			<td>Techcomp Limited</td>
			<td>NU5841</td>
			<td>For cell culture</td>
		</tr>
		<tr>
			<td>Dose calibrator</td>
			<td>Biodex</td>
			<td>&#160;N/A</td>
			<td>Atomlab 500</td>
		</tr>
		<tr>
			<td>DPBS (Dulbecco&#8217;s phosphate-buffered saline)</td>
			<td>Gibco</td>
			<td>14287072</td>
			<td>For cell wash and injection</td>
		</tr>
		<tr>
			<td>Eye lubricant</td>
			<td>Alcon Duratears</td>
			<td>&#160;N/A</td>
			<td>Sterile ocular lubricant ointment, 3.5 g</td>
		</tr>
		<tr>
			<td>Fetal bovine serum (FBS)</td>
			<td>Gibco</td>
			<td>A4766801</td>
			<td>Used for a broad range of cell types, especially sensitive cell lines</td>
		</tr>
		<tr>
			<td>Forceps (curved fine and straight blunt)</td>
			<td>RWD Life Science Co.,ltd</td>
			<td>F12012-10 &#38; F12011-13</td>
			<td>Animal surgery tool</td>
		</tr>
		<tr>
			<td>Heating pad</td>
			<td>ALA Scientific Instruments</td>
			<td>N/A</td>
			<td>Heat pad for mice during surgery</td>
		</tr>
		<tr>
			<td>Insulin syringe</td>
			<td>Terumo</td>
			<td>10ME2913</td>
			<td>1 mL insulin syringe with needle for radiotracer injections</td>
		</tr>
		<tr>
			<td>InterView fusion software</td>
			<td>Mediso</td>
			<td>Version 3.03</td>
			<td>Post-processing and image analysis software</td>
		</tr>
		<tr>
			<td>Inverted microscope</td>
			<td>Yu Lung Scientific Co., Ltd</td>
			<td>BM-209G</td>
			<td>For cells morphology visualization</td>
		</tr>
		<tr>
			<td>Isoflurane</td>
			<td>Chanelle Pharma</td>
			<td>&#160;N/A</td>
			<td>Iso-Vet, inhalation anesthetic, 250 mL</td>
		</tr>
		<tr>
			<td>Ketamine</td>
			<td>Alfasan International B.V.</td>
			<td>HK-37715</td>
			<td>Ketamine 10% injection solution, 10 mL&#160;</td>
		</tr>
		<tr>
			<td>Medical oxygen</td>
			<td>Linde HKO</td>
			<td>101-HR</td>
			<td>compressed gas, 99.5% purity</td>
		</tr>
		<tr>
			<td>nanoScan PET/MR Scanner</td>
			<td>Mediso</td>
			<td>&#160;N/A</td>
			<td>3 Tesla MR</td>
		</tr>
		<tr>
			<td>Needle holder</td>
			<td>RWD Life Science Co.,ltd</td>
			<td>F31026-12</td>
			<td>Animal surgery tool</td>
		</tr>
		<tr>
			<td>Nucline nanoScan software</td>
			<td>Mediso</td>
			<td>Version 3.0</td>
			<td>Scanner operating software</td>
		</tr>
		<tr>
			<td>Nylon Suture (6/0 and 5/0)</td>
			<td>Healthy Medical Company Ltd</td>
			<td>000524 &#38; 000526</td>
			<td>Animal surgery tool</td>
		</tr>
		<tr>
			<td>Penicillin- Streptomycin</td>
			<td>Gibco</td>
			<td>15140122</td>
			<td>Culture media for a final concentration of 50 to 100 I.U./mL penicillin and 50 to 100 &#181;g/mL streptomycin.</td>
		</tr>
		<tr>
			<td>Pentabarbital</td>
			<td>AlfaMedic</td>
			<td>13003</td>
			<td>Intraperitoneal injection (330 mg/kg) to induce cessation of breathing of mice</td>
		</tr>
		<tr>
			<td>Sharp scissors</td>
			<td>RWD Life Science Co.,ltd</td>
			<td>S14014-10</td>
			<td>Animal surgery tool</td>
		</tr>
		<tr>
			<td>Spring Scissors</td>
			<td>RWD Life Science Co.,ltd</td>
			<td>S11005-09</td>
			<td>Animal surgery tool</td>
		</tr>
		<tr>
			<td>Trypan Blue Solution, 0,4%</td>
			<td>Gibco</td>
			<td>15250061</td>
			<td>For cell counting</td>
		</tr>
		<tr>
			<td>Trypsin-ethylenediaminetetraacetic acid (EDTA, 0.25%), phenol red.</td>
			<td>Gibco</td>
			<td>25200072</td>
			<td>For cell digestion</td>
		</tr>
		<tr>
			<td>Xylazine</td>
			<td>Alfasan International B.V.</td>
			<td>HK-56179</td>
			<td>Xylazine 2% injection solution, 30 mL</td>
		</tr>
	</tbody>
</table>

non-invasive pet/mr imaging in an orthotopic mouse model of hepatocellular carcinoma

肝细胞癌（HCC）的临床前实验模型概括了人类疾病，是研究肿瘤发生和评估新治疗方法的重要工具。使用正电子发射断层扫描 （PET） 的非侵入性全身成像可在分子水平上实时提供对组织体内特征的关键见解。我们在这里提出了一个原位HCC异种移植物创建的方案，有和没有肝动脉结扎（HAL）诱导肿瘤缺氧，并使用[18 F]氟咪唑（[18 F]FMISO）和[18 F]氟脱氧葡萄糖（[18F]FDG）PET/磁共振（MR）成像评估其体内肿瘤代谢。使用缺氧标记物[18 F]FMISO可以很容易地观察到肿瘤缺氧，并且发现接受HAL的HCC小鼠的[18 F]FMISO摄取高于非HAL组，而[18F]FDG无法区分两组之间的肿瘤缺氧。HAL肿瘤在缺氧反应中也表现出更高水平的缺氧诱导因子（HIF）-1α表达。HAL肿瘤的定量显示，基于标准化价值摄取（SUV）方法的[18F] FMISO摄取增加了2.3倍。

为了获得适合连续原位植入的肿瘤阻滞，首先通过将DPBS（含有MHCC97L细胞）中的200μL细胞悬液皮下注射到裸鼠的下侧来产生稳定的克隆（图1A）。监测肿瘤生长，当肿瘤大小达到800-1000mm 3（注射后约4周）时，对小鼠实施安乐死，并将所得肿瘤块切成约1mm3片段，随后将肝原位植入另一批裸鼠（n = 6）。将小鼠随机分为两组：对照组（C1，n = 3）和肝动脉结扎术（...

Watch this Scientific Journal Video about Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma at JoVE.com

Non-Invasive PET/MR Imaging in an Orthotopic Mouse Model of Hepatocellular Carcinoma

在这里，我们提出了一种方案来创建有和没有肝动脉结扎的原位肝细胞癌异种移植物，并使用[18 F]氟咪唑（[18 F]FMISO）和[18 F]氟脱氧葡萄糖（[18F]FDG）对肿瘤缺氧进行非侵入性正电子发射断层扫描（PET）成像。

肝细胞癌原位小鼠模型中的无创PET/MR成像

Preclinical experimental models of hepatocellular carcinoma (HCC) that recapitulate human disease represent an important tool to study tumorigenesis and ...

我们的方法为肿瘤缺氧的无创成像创建了一个可靠的缺氧肝细胞癌模型，以研究体内HCC生物学，这可以更好地概括人类癌症的发展。我们可靠的HCC模型以及高度可生产的图像方法为人类HCC病理生理学的转化研究提供了重要平台。实时监测肿瘤缺氧可深入了解导致肿瘤侵袭性的分子变化，从而确定缺氧区域并预测对治疗的反应。虽然腹腔内手术在技术上具有挑战性，但充分的计划和动物护理将提高整体手术成功率和小动物的健康。首先，用70%乙醇对下侧左侧或右侧麻醉的6至8周龄小鼠的背侧区域进行灭菌。然后，用镊子提起皮肤，轻轻地将注射针插入下面。为避免在抽出针头期间注射的细胞悬液意外泄漏，请将针头沿皮下平面从穿刺部位推进 4 至 5 毫米。释放镊子后，小心地排出注射器的内容物，不要穿透另一侧。对皮下荷瘤小鼠实施安乐死后，用手术刀刀片在肿瘤部位的皮肤上切开一个切口。然后，切除肿瘤块并立即将其转移到培养基中。接下来，使用锋利的手术剪刀，将肿瘤块切成1立方毫米的小立方体，避开肿瘤块的核心区域。将所有立方体保存在培养基中。对于原位肝植入术，伸展麻醉小鼠的四肢并用胶带固定它们，以最大限度地暴露腹腹部和胸部。然后，用10%聚维酮碘溶液和70%乙醇对整个腹部皮肤进行消毒。接下来，首先使用锋利的剪刀进行剖腹手术，以进入腹膜，首先做一个 10 毫米的中线切口。然后，用镊子夹紧剑突并将其拉向头部，并使用肋下牵开器打开手术区域。使用湿纱布拭子，向上缩回正中和左肝叶。然后，将腹部外的肠子移到小鼠的左侧，并用湿纱布拭子覆盖它们。接下来，在放大灯下获得最佳可视化效果，对肝总动脉进行肝动脉结扎术，肝总动脉从胰头起源到肝蒂根部。用湿纱布拭子缩回中叶和左叶的背部，露出左叶。然后，使用无菌手术刀刀片，在左叶上做一个长约2毫米深的切口，并立即用无菌镊子将肿瘤碎片插入肝切口。在应用八字形缝合线之前，将肿瘤牢固地埋在肝脏中，在切口部位用6-0尼龙缝合线，以确保适当的肿瘤植入和止血。然后，用中断的5-0缝合线关闭腹部切口。将等分试样18-FMISO分配到管中，并用无菌盐水稀释，以确保100微升中的总活性浓度为18至20兆贝克勒尔。接下来，用1毫升注射器抽取溶液，并记录剂量校准器上显示的放射性和时间。记录受试者小鼠的体重后，小心地通过尾静脉注射制备的放射性示踪剂。为了说明衰减校正，记录注射器中的注射时间和残留活性。要开始PET采集，请通过执行侦察视图来定位扫描仪中的鼠标位置。调整鼠标床位置，以确保整个鼠标身体与躯干区域位于MR视野的中心。接下来，在研究列表窗口中选择 PET 采集。要使用侦察视图中的预定鼠标床位置，请选择先前采集的扫描范围。然后，单击“准备”以自动将鼠标床从MR移动到PET，然后单击“Go”开始采集。在放射性药物编辑器下选择合适的放射性核苷酸，然后输入注射剂量和时间的详细信息。此外，在“主题信息”菜单下，输入鼠标的重量。PET 扫描完成后，选择“准备”将鼠标从 PET 移动到 MR，然后单击“执行 MR 采集”。完成所有扫描后，通过选择表输出将成像床移回默认位置。本研究中使用的PET成像显示，在肝动脉结扎的小鼠中，18-FMISO的肿瘤摄取增加，但在对照小鼠中没有。然而，糖酵解标志物18F-FDG的肿瘤摄取在有和没有肝动脉结扎的小鼠中相似。使用这种标准化的基于摄取值的方法进行定量显示，肝动脉结扎小鼠肿瘤的18-FMISO摄取比对照小鼠高2.3倍。同样，肝动脉结扎小鼠肝脏中的18-FMISO摄取高于对照小鼠的肝脏。肝动脉结扎也导致肿瘤缺氧增强，肝动脉结扎小鼠肿瘤分泌物中缺氧诱导因子1-α表达水平高于对照小鼠。在18-FMISO PET成像后，可以进行测序以确定缺氧诱导的代谢基因的作用，从而产生新的治疗策略。该技术可用于准确监测肿瘤缺氧，从而可以快速评估治疗效果，并创建针对HCC缺氧途径的新治疗策略。

Research

JoVE Journal

Cancer Research

Fluorescent Orthotopic Mouse Model of Pancreatic Cancer

胰腺癌的荧光原位小鼠模型

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 ...

科研

癌症研究

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 ...

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

研究病毒载体在三维肝模型重新填充与人肝癌细胞株HepG2

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 ...

Isolation of Circulating Tumor Cells in an Orthotopic Mouse Model of Colorectal Cancer

在直肠癌原位小鼠模型中循环肿瘤细胞的分离

Despite the advantages of easy applicability and cost-effectiveness, subcutaneous mouse models have severe limitations and do not accurately simulate ...

An Oncogenic Hepatocyte-Induced Orthotopic Mouse Model of Hepatocellular Cancer Arising in the Setting of Hepatic Inflammation and Fibrosis

在肝炎症和纤维化环境中产生的肝细胞癌的致癌肝细胞诱导正交小鼠模型

The absence of a clinically relevant animal model addressing the typical immune characteristics of hepatocellular cancer (HCC) has significantly impeded ...

Ultrasound-Guided Orthotopic Implantation of Murine Pancreatic Ductal Adenocarcinoma

毛尿胰腺腺腺腺癌超声引导正射植入

The recent success of immune checkpoint blockade in melanoma and lung adenocarcinoma has galvanized the field of immuno-oncology as well as revealed the ...

Transradial Access Chemoembolization for Hepatocellular Carcinoma Patients

肝细胞癌患者的跨径向获取化疗

Transarterial chemoembolization (TACE) is the most common modality for treatment of hepatocellular carcinoma (HCC) at the intermediate stage. TACE is ...

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An Orthotopic Resectional Mouse Model of Pancreatic Cancer

胰腺癌矫形小鼠模型

There is a lack of satisfactory animal models to study adjuvant and/or neoadjuvant therapy in patients being considered for surgery of pancreatic cancer ...

Analysis of Liver Microenvironment During Early Progression of Non-Alcoholic Fatty Liver Disease-Associated Hepatocellular Carcinoma in Zebrafish

斑马鱼非酒精性脂肪肝相关肝细胞癌早期进展期间的肝脏微环境分析

Liver cancer is currently the third&#160;leading cause of cancer related death worldwide, and Hepatocellular Carcinoma (HCC) accounts for 75-90% of all ...

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

一种三维球体模型研究肝细胞癌肿瘤-基质相互作用

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