Name: gene regulation and targeted therapy in gastric cancer peritoneal metastasis: radiological findings from dual energy ct and pet/ct
Uploaded: 2018-01-22T14:00:00.000+00:00
Duration: 10 min 28 s
Description: Watch this Scientific Journal Video about Gene Regulation and Targeted Therapy in Gastric Cancer Peritoneal Metastasis: Radiological Findings from Dual Energy CT and PET/CT at JoVE.com

这项工作得到了自然科学基金 (No。U1532107) 与上海交通大学生物医学工程项目 (No。YG2014MS53)。作者感谢叶剑英李和燕神对 DECT 和 PET/CT 成像方法的有益评价和技术支持。

这项工作是严格按照上海交通大学实验动物保护和使用指南所制定的标准进行的, 并获得瑞金医院实验室动物伦理委员会批准。
1. 胃癌腹膜转移动物模型
<ol><li>将一个适度分化的 SGC-7901 人胃癌细胞系分为 SGC-7901/sFRP1 组和 SGC-7901/vector 组。培养两组细胞分别在 RPMI 1640 补充10% 胎牛血清, 100 单位/毫升链霉素, 和100µg/毫升青霉素在37° c 的湿化气氛与 5% CO2。</li>
	<li>使用4-6 周老的女性裸 balb/c/c 裸鼠与体重25至 30 g. 在动物设施中的特定无病原体条件下放置动物。</li>
	<li>将小鼠随机分为 sFPR1 过度表达组和 sFPR1 空负荷组。 注: 每组有十裸鼠;二十只小鼠随机分为 TGF-β1治疗组和 TGF-β1对照组, 每只动物笼有多达5只裸鼠。</li>
	<li>建立 sFPR1-overexpression 腹膜转移移植模型组, 通过管理150µL (2 x 106细胞/毫升) 通过腹腔 SGC-7901/sFRP1 细胞的悬浮;SGC-7901/vector 单元被管理以建立空加载组。 注意: 使用例计数方法确定单元格的浓度20。</li>
	<li>
建立腹膜转移移植模型组通过管理150µL (2 x 106细胞/毫升) 悬浮 SGC-7901 细胞通过腹腔。管理的目标治疗 TGF-β1抑制剂 SB431542 后两个星期的增长, 经腹腔注射100µL/10 克体重每隔一天对小鼠的治疗组。
	<ol>
<li>对对照组的小鼠进行生理盐水的相同剂量管理。</li>
	</ol>
</li>
	<li>执行 DECT 和 PET/CT 扫描1天前治疗和1天, 7 天, 14 天, 和21天后治疗。</li>
</ol>2. 腹膜转移动物模型的 DECT
注: 在双能量 CT 扫描仪上实现了动物成像实验 (见材料表)。根据前人的研究, 我们建立了相关的 DECT 成像协议。
<ol><li>DECT 成像协议的设置
	<ol>
<li>在映像控制台计算机上, 选择 "协议管理" 图标进入下一个接口, 然后单击 "协议管理" 选项以查看协议管理屏幕。</li>
		<li>在 "用户协议" 界面中, 选择腹部区域进入腹部协议列表。</li>
		<li>单击协议列表中的空白区域, 然后选择 "新建" 按钮以键入新协议的名称: "动物 DECT 扫描"。按键盘上的 "Enter" 键, 然后在弹出窗口中选择 "侦察" 按钮, 单击 "确定" 以设置侦察系列 (第一个系列)。</li>
		<li>选择 "XY" 模式的 "解剖参考点" 和 "头第一仰卧位" 的 ' 病人方向 '。单击 "自动转移" 选项, 然后选择要上载图像系列的工作站位置。在系列描述中命名 "侦察阶段"。</li>
		<li>在 "查看编辑" 屏幕中, 确保相关的扫描参数设置如下: 开始位置 "和" 结束位置 "选项分别设置在" S50 "和" I50 "," KV "在" 100 "," mA "在" 80 "," 90 ° "为" 横向侦察位置 "," 0 ° "为" AP 侦察位置", 和" 侦察员 WW "在" 400/40 "。</li>
		<li>接下来, 创建非增强扫描的第二个系列。点击 "创建新系列", 在弹出窗口中选择 "轴向" 和 "创建后" 图标。</li>
		<li>将系列命名为 "-c 阶段", 并打开 "显示本地化"。在扫描类型接口中, 选择 "螺旋" 扫描类型和 0.5 s 为 "旋转时间", 单击 "粗速度" 选项设置参数 (探测器覆盖范围在40毫米, 螺旋厚度在0.625 毫米, 音高和速度在 0.516:1/20.62, 旋转时间在0.5 秒) 在弹出窗口, 间隔在0.625 毫米, 龙门乳头在 0, SFOV 选择小体, kV 在 100, 点击 "ma" 然后键入600为手动 ma。</li>
		<li>点击 "侦察参数" 图标, 打开 "侦察选项" 弹出窗口。在侦察模式中选择 "加号";单击 "切片" 图标, 在 "ASiR 安装屏幕" 中选择 "ss50 切片 50%" 模式。设置其余参数如下: DFOV 在25厘米, R/L 和一个/P 中心在0厘米, 侦察类型选择 "Stnd", 矩阵大小在512。</li>
		<li>通过重复步骤2.1.8 为增强扫描创建第三个扫描系列, 将其命名为 "+ C QC 阶段" 并打开 "显示定位器";第一组是动脉相系列的设置。</li>
		<li>在 "扫描类型" 界面中, 点击 "(宝石光谱成像)", 并选择 "螺旋" 扫描类型, 选择协议 "GSI-52" 在 ' 腹式预置选择 ' 窗口。根据非增强扫描设置起始位置和结束位置。<ol>
<li>点击 "侦察参数" 图标, 打开 "侦察选项" 弹出窗口。在侦察模式中, 选择 "加号", 然后在 "选项" 中单击 "QC";其余参数与步骤2.1.8 相同。</li>
		</ol>
</li>
		<li>单击 "R2" 图标, 并在 "启用侦察" 选项卡中选择 "是". 选择 "厚度" 在0.625 和类型0.625 为 "间隔"。打开 "侦察选项" 弹出窗口, 在侦察模式选择 "加号";在 "选择" 选项中, 单击 "单声道" 并将凯文设置为70凯文;在 ASiR 设置窗口中, 选择 "GS40 40%" 模式用于 ASiR 安装。其余参数设置为步骤2.1.8。将此步骤命名为 "+ C 70keV 阶段"。</li>
		<li>单击 "R3" 图标, 并在 "启用侦察" 选项卡中选择 "是". 将 "厚度" 设置为 1.25, 类型0.625 为 "间隔"。打开 "侦察选项" 弹出窗口, 在侦察模式中选择 "加号" 和 "IQ 增强";在 "单声道" 选项中, 单击 "单音", 将凯文设置为70凯文, 然后点击 "数据文件";确保 ASiR 安装符合步骤13。其余参数设置为步骤2.1.8。将其命名为 "+ C 单阶段"。</li>
		<li>单击 "添加组" 以创建两个扫描组, 分别表示门户阶段和延迟阶段。确保每个扫描阶段的 "起始位置" 和 "结束位置" 范围一致, 其余参数与动脉阶段相同。在 "准备组" 中键入延迟时间: 第一组 (动脉阶段) 在0秒, 第二组 (门阶段) 在8秒, 第三组 (延迟阶段) 在十六年代。</li>
		<li>在完成所有设置后, 单击 "接受" 选项以保存协议。</li>
	</ol></li>
	<li>DECT 成像过程
	<ol>
<li>在每次扫描前随机从治疗和对照组中选择裸鼠。将选定的动物置于新的笼子中, 并分别标记它们。</li>
		<li>用清水将小鼠快速带入4小时, 但没有食物或被褥。</li>
		<li>在扫描前1小时将实验小鼠从动物实验中心移走, 并确保在扫描开始之前将老鼠置于一个新的温暖环境中。</li>
		<li>麻醉所有实验小鼠腹腔注射2.5% 戊巴比妥钠 (1.0 毫升/千克体重) 前 DECT 扫描成像, 并确认麻醉深度的脚趾捏反射。在麻醉的时候用眼药膏来防止干燥。 注: 在注射药物时, 确保每个裸鼠头部处于较低位置, 从而减少对内脏器官的损伤。注意注射部位和注射深度。将注射器的尖端放在45°角, 放在右/左下腹内, 并确保针的深度是这样, 以避免注射到肠道和其他器官。</li>
		<li>单击 "新患者" 图标, 输入有关鼠标的基本信息, 包括其患者 ID 和名称。在 "用户协议" 中, 单击 "腹部协议" 并选择 "动物 DECT 扫描" 协议进入操作界面。</li>
		<li>一旦引起麻醉, 将每个鼠标移动到一个动物夹具平台的仰卧位, 并修复它的尾巴与磁带, 以确保它不弯曲。用酒精对尾部进行消毒, 随后将造影剂注入尾部静脉。</li>
		<li>移动 CT 扫描床, 使外部定位线激光位于动物的下腹部。当定位完成时, 单击 "重置" 按钮。 注意: 在动物的下腹部放置外部定位线, 确保动物尽可能地位于机器的外部, 便于造影剂的管理进入尾部静脉。</li>
		<li>单击 "确认" 图标, 并按照键盘上按钮的闪烁顺序完成侦察扫描。在完成侦察扫描后, 选择 "下一序列" 图标, 并输入用于非增强扫描的界面。</li>
		<li>在右屏幕上, 设置 "开始位置" 和 "结束位置" 的侦察员视图, 以定义扫描范围。在 "横向侦察" 和 "AP 侦察" 中保持相同的范围, 并覆盖整个动物的身体体积。</li>
		<li>单击 "确认" 图标, 并按照键盘上按钮的闪烁顺序完成侦察扫描。</li>
		<li>通过尾静脉注射每只帕 0.2 mL/100 g 的小鼠。 注: 我们选择手动管理对比剂, 保持注塑速度尽可能稳定。这是最有利于捕捉早期增强肿瘤在成像。</li>
		<li>单击 "下一系列" 以执行增强扫描。根据非增强扫描设置 "开始位置" 和 "结束位置"。单击 "确认" 图标, 并按照键盘上按钮的闪烁顺序完成动态增强扫描、includingarterial 阶段、门户阶段和延迟阶段。 注: 在注射造影剂后立即单击 "确认" 图标开始扫描。这对于增强扫描以确保捕获动脉相的最佳图像至关重要。但是, 单击 "确认" 图标后的一些延迟时间可以确保实验人员从扫描室安全地撤回。</li>
		<li>点击 "结束考试", 在扫描完成后退出扫描界面;图像系列将自动上载到工作站。</li>
		<li>完成扫描后, 将动物放到一个空笼子里, 观察它们, 直到它们恢复知觉。不要让动物无人看管, 直到它恢复了足够的意识来维持胸骨卧床。然后把老鼠转移到一个干净的动物房间。</li>
	</ol></li>
	<li>后 DECT 成像分析
	<ol>
<li>在 DECT workstationinterface 上找到鼠标系列 (请参阅材料表) 并选择 "+ C 单阶段" 系列列表。打开 "加密卷查看器", 并从 "协议管理器" 接口中选择 "一般" 协议。</li>
		<li>单击图像视左上角的 "视图类型" 活动注释, 然后从下拉菜单中选择 "日冕" 方向。</li>
		<li>对于一个图像视区, 单击左上角的 "卷 1" 活动注释, 然后从下拉菜单中选择 "单声道" 卷。同样, 在另一个图像视区中, 选择 "碘 (水)" 卷。单击并按住鼠标左键, 将 "碘 (水)" 视区中的图像拖动到 "单声道", 并检查 "混合视图" 框以获得颜色融合图像。</li>
		<li>单击并从 "图像滚动" 图标的中心拖动以观察图像。将显示阳性结果的图像保存为彩色融合图像。</li>
	</ol></li>
</ol>3. PET/CT 在腹膜转移动物模型中的研究
注: 见所用的 PET/CT 成像仪的材料表。我们根据本文21创建了相关的 PET/CT 成像协议。
<ol><li>
设置微 PET/CT 成像协议
	<ol>
<li>对于全身 CT 扫描, 设置电流在500µA, 电压在80伏, 曝光时间为200毫秒, 240 步为240旋转。对于 x 射线探测器, 选择分辨率在 "低系统放大" 与78毫米轴向成像场和单人床模式。采用 "普通锥梁重构" 方法, 选择 "实时重建" 选项, 使上位机能够与专用的实时重建计算机 (眼镜蛇) 连接, 启动任务。</li>
		<li>对于 PET 的采集, 在 "获取时间" 选项设置 "固定扫描时间" 到 600s (10 分钟)。设置 "研究同位素" 到 F-18 和 "能量水平" 到350-650 凯文。</li>
		<li>为了生成 PET 直方图, 将 "动态帧" 设置为 "黑色", 以便在整个持续时间内将数据作为一个帧来处理, 以实现静态扫描。将直方图类型设置为 "3D", 并选择 "无散射校正" 选项。</li>
		<li>对于 pet 重建, 使用 OSEM3D 算法重建图像, 然后是由 PET/CT workstationsoftware 提供的地图或快速映射22 (请参阅材料表)。</li>
	</ol>
</li>
	<li>
PET/CT 成像前的制备
	<ol>
<li>快速进行4小时 DECT 实验的小鼠, 在成像前30分钟将小鼠转移到新的动物笼中。</li>
		<li>称量小鼠并记录它们的重量。</li>
		<li>按照研究所的安全程序获取和携带含有放射性物质 (RAM) 的包裹。使用防护罩进行18f-葡萄糖 (5 mCi), 并用剂量校验仪测量总的18f-葡萄糖的放射性。</li>
		<li>用生理盐水稀释18F-葡萄糖对小鼠注射液的适当放射性。 注: 18F-葡萄糖的稀释活性应为每只老鼠提供100-200 µCi/100 µL。<ol>
<li>绘制200µL 18F-葡萄糖溶液成1毫升注射器。用剂量校验仪测量整个注射器的放射性, 并记录18F-葡萄糖的准备时间。</li>
		</ol>
</li>
		<li>使用200µL 18f-葡萄糖溶液通过尾部静脉注射路径注入每个鼠标, 并记录18f-葡萄糖注射液时间。注射完所有小鼠后, 立即用剂量校验仪测量注射器的残余放射性, 记录注射完成后测量的时间。</li>
		<li>按以下公式计算每个鼠标的插入的18F-葡萄糖活性: 注射活性 (µCi) = 注射前注射器活性在注射器注入后的活性。</li>
	</ol>
</li>
	<li>
PET/CT 成像过程
	<ol>
<li>将动物放入麻醉诱导室;麻醉小鼠使用吸入3% 异氟醚在氧后完成了18F-葡萄糖注射液。 注意: 遵照所有动物福利准则进行操作;使用加热垫使小鼠保持温暖。用眼药膏预防麻醉时的干燥。</li>
		<li>一旦引起麻醉, 将鼠标移动到微 CT 扫描床上, 同时保持持续的麻醉和变暖。将鼠标的头部放在一个锥面掩模内, 在氧气中连续输送异氟醚 (2%), 流速为2升/分. 将鼠标放在仰卧位, 以确保其姿态与 DECT 扫描中的姿势一致。</li>
		<li>把动物移到宠物/CT 扫描仪的入口处, 单击工具栏查看器中的 "激光" 图标, 并使用触摸板控制界面移动床, 使鼠标的腹部位于 PET 和 CT 场的中心 (视) 扫描过程中。在 "激光对准" 窗口中, 选择 "第一扫描类型" 作为 "CT 扫描", "工作流中包含的 PET 捕获" 作为选择。</li>
		<li>打开 "侦察员视图" 窗口, 并获得一个侦察兵查看 X 射线照相。调整动物床的位置, 使 CT 的中心视野位于鼠标身体的中心。</li>
		<li>选择以前建立的协议 (在步骤3.1 中)。在弹出窗口中输入要映像的鼠标数 (先后), 然后单击 "设置" 选项, 然后输入权重。然后再次单击 "安装" 选项, 然后按照弹出窗口说明完成安装。</li>
		<li>单击 "启动工作流" 图标开始扫描。</li>
		<li>在所有扫描完成后, 评估获得的 CT 和 PET 图像的质量。将数据通过网络传送到后成像分析中进行进一步的研究。 注意: 调整图像的窗口宽度和窗口级别, 以确保正确显示各器官的对比度。检查图像中各器官的分辨率, 确认成像质量。</li>
		<li>将动物从成像仪中取出, 立即安乐颈椎脱位。连续使用下一动物的成像系统。</li>
	</ol>
</li>
	<li>
后 PET/CT 成像分析
	<ol>
<li>打开 pet/ct 工作站软件, 将 ct 和 pet 图像系列数据导入软件中。在 "注册" 窗口中, 单击 "一般分析" 选项, 将 ct 和 pet 图像进行注册, 并在 "查看" 窗口下选择 "天空" 模型, 以显示 ct 和 pet 图像之间的完美匹配。</li>
		<li>在 "感兴趣区域 (ROI) 量化" 窗口中, 通过 co 注册图像提供的参考来识别腹膜结节。</li>
		<li>在 "感兴趣区域 (roi) 量化" 窗口中, 用工具在熔融图像上绘制 roi, 编辑 roi 的大小和形状, 记录 SUV 的max值, 然后输出并保存所选的合并图像。</li>
	</ol>
</li>
</ol>

<ol>
	<li>Ferlay, 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=Cancer+incidence+and+mortality+worldwide:+sources,+methods+and+major+patterns+in+GLOBOCAN+2012.">Cancer incidence and mortality worldwide: sources, methods and major patterns in GLOBOCAN 2012.</a> Int J Cancer. 136 (5), 359-386 (2015).</li><li>Kobayashi, D., Kodera, Y. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Intraperitoneal+chemotherapy+for+gastric+cancer+with+peritoneal+metastasis.">Intraperitoneal chemotherapy for gastric cancer with peritoneal metastasis.</a> Gastric Cancer. 20, 111-121 (2017).</li><li>Gu, W., Li, X., Wang, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=miR-139+regulates+the+proliferation+and+invasion+of+hepatocellular+carcinoma+through+the+WNT/TCF-4+pathway.">miR-139 regulates the proliferation and invasion of hepatocellular carcinoma through the WNT/TCF-4 pathway.</a> Oncol Rep. 31 (1), 397-404 (2014).</li><li>Sugai, T., 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+analysis+of+gastric+differentiated-type+intramucosal+and+submucosal+cancers.">Molecular analysis of gastric differentiated-type intramucosal and submucosal cancers.</a> Int J Cancer. 127 (11), 2500-2509 (2010).</li><li>Shi, Y., He, B., You, L., Jablons, D. M. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Roles+of+secreted+frizzled-related+proteins+in+cancer.">Roles of secreted frizzled-related proteins in cancer.</a> Acta Pharmacol Sin. 28 (9), 1499-1504 (2007).</li><li>Amin, N., Vincan, E. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+Wnt+signaling+pathways+and+cell+adhesion.">The Wnt signaling pathways and cell adhesion.</a> Front Biosci (Landmark Ed). 17, 784-804 (2012).</li><li>Jones, S. E., Jomary, C. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Secreted+Frizzled-related+proteins:+searching+for+relationships+and+patterns.">Secreted Frizzled-related proteins: searching for relationships and patterns.</a> Bioessays. 24 (9), 811-820 (2002).</li><li>Qu, 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=High+levels+of+secreted+frizzled-related+protein+1+correlate+with+poor+prognosis+and+promote+tumourigenesis+in+gastric+cancer.">High levels of secreted frizzled-related protein 1 correlate with poor prognosis and promote tumourigenesis in gastric cancer.</a> Eur J Cancer. 49 (17), 3718-3728 (2013).</li><li>Pan, 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=Determining+gastric+cancer+resectability+by+dynamic+MDCT.">Determining gastric cancer resectability by dynamic MDCT.</a> Eur Radiol. 20 (3), 613-620 (2010).</li><li>Pan, 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=Gastric+cancer+staging+with+dual+energy+spectral+CT+imaging.">Gastric cancer staging with dual energy spectral CT imaging.</a> PLoS One. 8 (2), 53651 (2013).</li><li>Kim, M. J., Hong, J. H., Park, E. S., Byun, J. H. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Gastric+metastasis+from+primary+lung+adenocarcinoma+mimicking+primary+gastric+cancer.">Gastric metastasis from primary lung adenocarcinoma mimicking primary gastric cancer.</a> World J Gastrointest Oncol. 7 (3), 12-16 (2015).</li><li>Maeda, H., Kobayashi, M., Sakamoto, J. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Evaluation+and+treatment+of+malignant+ascites+secondary+to+gastric+cancer.">Evaluation and treatment of malignant ascites secondary to gastric cancer.</a> World J Gastroenterol. 21 (39), 10936-10947 (2015).</li><li>Bensinger, S. J., Christofk, H. R. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=New+aspects+of+the+Warburg+effect+in+cancer+cell+biology.">New aspects of the Warburg effect in cancer cell biology.</a> Semin Cell Dev Biol. 23 (4), 352-361 (2012).</li><li>Smyth, 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=A+prospective+evaluation+of+the+utility+of+2-deoxy-2-[(18)+F]fluoro-D-glucose+positron+emission+tomography+and+computed+tomography+in+staging+locally+advanced+gastric+cancer.">A prospective evaluation of the utility of 2-deoxy-2-[(18) F]fluoro-D-glucose positron emission tomography and computed tomography in staging locally advanced gastric cancer.</a> Cancer. 118 (22), 5481-5488 (2012).</li><li>Oka, S., Uramoto, H., Shimokawa, H., Iwanami, T., Tanaka, F. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=The+expression+of+Ki-67,+but+not+proliferating+cell+nuclear+antigen,+predicts+poor+disease+free+survival+in+patients+with+adenocarcinoma+of+the+lung.">The expression of Ki-67, but not proliferating cell nuclear antigen, predicts poor disease free survival in patients with adenocarcinoma of the lung.</a> Anticancer Res. 31 (12), 4277-4282 (2011).</li><li>Zhao, C. H., Bu, X. M., Zhang, N. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Hypermethylation+and+aberrant+expression+of+Wnt+antagonist+secreted+frizzled-related+protein+1+in+gastric+cancer.">Hypermethylation and aberrant expression of Wnt antagonist secreted frizzled-related protein 1 in gastric cancer.</a> World J Gastroenterol. 13 (15), 2214-2217 (2007).</li><li>Cheson, B. D. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Role+of+functional+imaging+in+the+management+of+lymphoma.">Role of functional imaging in the management of lymphoma.</a> J Clin Oncol. 29 (14), 1844-1854 (2011).</li><li>Fuster, 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=Preoperative+staging+of+large+primary+breast+cancer+with+[18F]fluorodeoxyglucose+positron+emission+tomography/computed+tomography+compared+with+conventional+imaging+procedures.">Preoperative staging of large primary breast cancer with [18F]fluorodeoxyglucose positron emission tomography/computed tomography compared with conventional imaging procedures.</a> J Clin Oncol. 26 (29), 4746-4751 (2008).</li><li>Lin, 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=Secreted+frizzled-related+protein+1+overexpression+in+gastric+cancer:+Relationship+with+radiological+findings+of+dual-energy+spectral+CT+and+PET-CT.">Secreted frizzled-related protein 1 overexpression in gastric cancer: Relationship with radiological findings of dual-energy spectral CT and PET-CT.</a> Scientific Reports. 7, 42020 (2017).</li><li>Cadena-Herrera, 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=Validation+of+three+viable-cell+counting+methods:+Manual,+semi-automated,+andautomated.">Validation of three viable-cell counting methods: Manual, semi-automated, andautomated.</a> Biotechnol Rep (Amst). 7, 9-16 (2015).</li><li>Wang, X., Minze, L. J., Shi, Z. Z. <a target="_blank" href="http://www.ncbi.nlm.nih.gov/entrez/query.fcgi?db=PubMed&cmd=Search&doptcmdl=Citation&defaultField=Title+Word&term=Functional+imaging+of+brown+fat+in+mice+with+18F-FDG+micro-PET/CT.">Functional imaging of brown fat in mice with 18F-FDG micro-PET/CT.</a> J Vis Exp. (69), (2012).</li><li>Grootjans, 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=Performance+of+3DOSEM+and+MAP+algorithms+for+reconstructing+low+count+SPECT+acquisitions.">Performance of 3DOSEM and MAP algorithms for reconstructing low count SPECT acquisitions.</a> Z Med Phys. 26 (4), 311-322 (2016).</li><li>Chang, H. R., 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=Improving+gastric+cancer+preclinical+studies+using+diverse+in+vitro+and+in+vivo+model+systems.">Improving gastric cancer preclinical studies using diverse in vitro and in vivo model systems.</a> BMC Cancer. 16, 200 (2016).</li><li>Chang, H. R., 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=HNF4alpha+is+a+therapeutic+target+that+links+AMPK+to+WNT+signalling+in+early-stage+gastric+cancer.">HNF4alpha is a therapeutic target that links AMPK to WNT signalling in early-stage gastric cancer.</a> Gut. 65 (1), 19-32 (2016).</li><li>Zheng, H. 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=BTG1+expression+correlates+with+pathogenesis,+aggressive+behaviors+and+prognosis+of+gastric+cancer:+a+potential+target+for+gene+therapy.">BTG1 expression correlates with pathogenesis, aggressive behaviors and prognosis of gastric cancer: a potential target for gene therapy.</a> Oncotarget. 6 (23), 19685-19705 (2015).</li><li>Yamada, A., Oguchi, K., Fukushima, M., Imai, Y., Kadoya, M. <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+2-deoxy-2-[18F]fluoro-D-glucose+positron+emission+tomography+in+gastric+carcinoma:+relation+to+histological+subtypes+depth+of+tumor+invasion,+and+glucose+transporter-1+expression.">Evaluation of 2-deoxy-2-[18F]fluoro-D-glucose positron emission tomography in gastric carcinoma: relation to histological subtypes depth of tumor invasion, and glucose transporter-1 expression.</a> Ann Nucl Med. 20 (9), 597-604 (2006).</li><li>Hirose, 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=Relationship+between+2-deoxy-2-[(18)F]-fluoro-d-glucose+uptake+and+clinicopathological+factors+in+patients+with+diffuse+large+B-cell+lymphoma.">Relationship between 2-deoxy-2-[(18)F]-fluoro-d-glucose uptake and clinicopathological factors in patients with diffuse large B-cell lymphoma.</a> Leuk Lymphoma. 55 (3), 520-525 (2014).</li><li>Tchou, 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=Degree+of+tumor+FDG+uptake+correlates+with+proliferation+index+in+triple+negative+breast+cancer.">Degree of tumor FDG uptake correlates with proliferation index in triple negative breast cancer.</a> Mol Imaging Biol. 12 (6), 657-662 (2010).</li><li>Coleman, R. 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=Concurrent+PET/CT+with+an+integrated+imaging+system:+intersociety+dialogue+from+the+Joint+Working+Group+of+the+American+College+of+Radiology+the+Society+of+Nuclear+Medicine,+and+the+Society+of+Computed+Body+Tomography+and+Magnetic+Resonance.">Concurrent PET/CT with an integrated imaging system: intersociety dialogue from the Joint Working Group of the American College of Radiology the Society of Nuclear Medicine, and the Society of Computed Body Tomography and Magnetic Resonance.</a> J Am Coll Radiol. 2 (7), 568-584 (2005).</li><li>Brepoels, 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=Effect+of+corticosteroids+on+18F-FDG+uptake+in+tumor+lesions+after+chemotherapy.">Effect of corticosteroids on 18F-FDG uptake in tumor lesions after chemotherapy.</a> J Nucl Med. 48 (3), 390-397 (2007).</li><li>Spaepen, 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=[18)F]FDG+PET+monitoring+of+tumour+response+to+chemotherapy:+does+[(18)F]FDG+uptake+correlate+with+the+viable+tumour+cell+fraction.">[18)F]FDG PET monitoring of tumour response to chemotherapy: does [(18)F]FDG uptake correlate with the viable tumour cell fraction.</a> Eur J Nucl Med Mol Imaging. 30 (5), 682-688 (2003).</li></ol>

作者没有利益冲突声明。

动物模型在胃癌的分子机制研究中得到了广泛的应用, 并尝试了各种治疗策略23,24,25。在这项研究中, 我们描述了一个详细的协议, 胃癌腹膜转移裸鼠模型, 使用 DECT 和 PET/CT 图像胃肿瘤实时识别肿瘤细胞增殖, 并监测腹膜转移和胃癌动物模型的治疗干预反应。这种方法可以使参与研究胃癌的分子机制的研究人员, 或实验成像肿瘤, 建立更完整的和精确的计划。此外, 我们描述了 DECT 和 PET/CT 设备的使用, 可以作为发现肿瘤影像学表现与基因调控和靶向治疗的平台。因此, 这种方法可以被科学家用来理解和探索癌症复发和进展的生物学过程。我们已经证明, 非侵入性成像模式可以检测出增加肿瘤的基因过度表达的 DECT 和 PET/CT 阳性结果. 同时, 靶向治疗干预后腹膜转移抑制剂在 PET/CT 上显示负葡萄糖吸收性能。SUVmax 瘤结节呈下降趋势, 延长治疗周期。
在我们的研究中, 我们利用葡萄糖摄取的变化作为评估指标的治疗效果的腹膜转移。已经提出了巨大的努力, 以表明葡萄糖摄取与肿瘤侵袭性相关的26。进展性胃癌, 以浸润深度、淋巴渗透、血管浸润和肿瘤大小为代表, 表现出较高的葡萄糖摄取量27。在定量评价方面, 研究表明, SUVmax与各种恶性肿瘤的增殖呈正相关,15,28。我们的结果表明, 与对照组相比, sFRP1 过度表达积极诱导明显较大的结节, 显著提高和较高的葡萄糖摄取率在腹膜肿瘤, 如在图 1中证明。此外, SUVmax 在靶向治疗组中表现出明显的改变趋势, 对照组无变化。这些结果显示在图 3中, 其葡萄糖摄取量减少, 治疗组的 SUVmax 接近 0.83, 而对照组中的小鼠则不变的 SUVmax 值接近 1.26 (图4).我们的研究结果表明, 非侵入性成像技术, 如 DECT 和 PET/CT, 提供了利用图像技术来评估肿瘤细胞分子水平的信息的可能性, 并证明了结合 DECT 的应用的有效性和 PET/CT 提供一个可行的, 重现性的, 非侵入的成像策略, 以监测肿瘤诱导的基因调控胃癌的研究。由于葡萄糖摄取与肿瘤侵袭性相关26, 利用 PET/CT 成像评估肿瘤浸润和治疗的程度是可行的。
在以往的研究中, 我们发现注射时间和方法会影响 DECT 扫描的成像结果。随着小鼠血液循环的快速更改, 腹膜转移的增加主要发生在动脉期, 如果扫描开始时间明显晚于注射时, 腹膜转移结节可能不会完全显示。在检查后, 裸鼠应放置在一个温暖和清洁的环境中, 让小鼠在 pet/ct 成像前休息12小时, 以避免在 pet/ct 成像中使用过量的残留造影剂对结核的摄取产生影响18F-葡萄糖身体.在葡萄糖的制备和注射过程中应注意注射时间和活动。PET/CT 的最佳活性浓度为 100-200µCi/100µL, 每次注射均为18。浓度过高可能会加重循环系统负担, 导致小鼠死亡, 而浓度过低则会影响肿瘤摄取成像。因此, 确保18F-葡萄糖的配置的效率和准确性至关重要。确保裸鼠的 PET/CT 成像位置与 DECT 成像相一致, 以利于肿瘤图像的匹配。
我们的研究有几个限制。DECT 的有限分辨率可能会导致可见性的负面表现, 因为一些腹膜肿瘤可能表现得不够大。众所周知, PET/CT 的特异性低, 缺乏解剖定位, 化疗药物干预诱导的肿瘤细胞凋亡和坏死可能影响18F-葡萄糖摄取量29,30.此外, 肠道循环正常的生理活动和18F-葡萄糖在输尿管和膀胱中的保留可能有助于假阳性出现在 PET/CT 图像31。裸鼠腹膜转移过程中的模型难以检测和监测, 因此在治疗干预和影像学实验中选择合适的时间尤为重要。因此, 小肿瘤的早期诊断仍然是一个亟待解决的问题。
最后, 我们描述了一种利用 DECT 和 PET/CT 成像技术的方法, 用于准确检测和评价靶向治疗的疗效。我们的结果表明, 非侵入性成像使用所述的协议允许监测和评估腹膜转移进展使用动物模型。该方法的应用将很容易适应临床前研究, 旨在发现胃癌腹膜转移, 这可能是有用的评估疾病诊断或治疗模式。

胃癌 (GC) 仍然是第四最常见的恶性肿瘤, 也是全球癌症死亡率的第二大原因1。虽然胃癌的诊断和治疗的准确性有了很大的提高, 但腹膜转移是胃癌预后和复发的最关键因素, 是术后死亡的决定性指标2。人们普遍认为腹膜传播是一种危及生命的转移方式, 其中, 一旦建立腹膜播散, 疾病就无法控制, 病人的预后较差。因此, 胃癌腹膜转移的检测和疗效评价是临床应用的关键。
胃癌发病率和死亡率的上升促使研究人员识别其分子机制。高表达的基因, 如分泌的卷曲相关蛋白 1 (sFRP1) 可能导致激活的信号通路在早期阶段的胃癌, 促进肿瘤的生长, 增殖, 分化和凋亡的过程中3,4,5,6,7. sFRP1-overexpression 细胞显示 TGFβ的表达增加, 其下游目标, 和 TGFβ介导的急诊室8。以往的研究表明, TGF-β1水平与腹膜转移和胃癌 TNM 分期相关。我们描述了 sFRP1 过度表达和 TGF-β1抑制的癌细胞增殖的变化, 并建立了腹膜转移的动物模型, 以显示肿瘤在基因调控作用下的影像表现。
动物模型的胃癌是必不可少的工具, 研究肿瘤的发展和试验的各种治疗策略, 而不必牺牲动物。动物模型在研究肿瘤和起源细胞的形成机制、确定肿瘤干细胞的存在以及研究各种新的治疗策略方面证明是有用的。因此, 实时无创技术可以准确描述胃肿瘤的发展和治疗的肿瘤反应, 从而确定裸鼠腹膜转移结节的发展, 并监测肿瘤反应的各种实验和治疗干预。
目前, 多探测器 CT (MDCT) 在胃癌 TNM 分期中起着重要作用, 对预测肿瘤切除术后9有一定的参考价值。然而, 经病理证实的胃癌患者的影像学研究主要基于形态学。DECT 成像扩展参数, 以反映功能信息, 提供单色图像, 并可能有助于提高 N 分期准确性胃癌。此外, 这项技术将有助于获得材料分解图像, 这可能有助于区分分化和未分化的胃癌, 并之间转移和非转移淋巴结10.随着 DECT 的引入, CT 的功能成像方面也被添加到临床应用中, 有助于评价疗效和预测患者预后11,12,13。PET/CT 是一种有用的成像技术的检测和分期胃癌, 并能评估复发的肿瘤有效14。肿瘤细胞增殖和血管生成都被认为是必要的, 在开发一个可检测的肿瘤15, 肿瘤结节显示了一个积极的性能与更高的 SUVmax的 PET/CT. 基于对好氧的偏好糖酵解, 18F-葡萄糖的模拟, 已被利用作为一个有希望的示踪剂的诊断恶性肿瘤, 结合 PET/CT16。这种方法依赖于肿瘤组织的快速葡萄糖消耗, 并具有广泛的临床应用, 包括协助检测、分期和评估肿瘤的预后, 以及监测肿瘤对治疗的反应17,18. 由于非侵入性的方法, DECT 和 PET/CT 已被用于诊断恶性肿瘤和评估肿瘤对各种疗法的反应。
我们的小组一直使用这种非侵入性成像方法与 DECT 和 PET/CT 扫描仪, 以检测和监测的过程中的肿瘤生长和转移的活鼠19。我们探讨了 sFRP1-overexpression 诱导的胃癌细胞的影像学表现在体内使用裸鼠, 与 DECT 和 PET/CT, 并描述了 SUVmax值的变化后, TGF-β1抑制剂的靶向治疗, 以确认研究了基因诱导后腹膜肿瘤结节的发展, 并对实验性治疗中肿瘤结节的变化进行了探讨。本文对小鼠胃肿瘤腹膜转移的建模、DECT 和 PET/CT 的检测与监测提供了详细的方法。

<table><tbody><tr><td>Iohexol</td><td>BEJING BEILU PHARMACEUTICAL CO,LTD</td><td>NMPN:H20053800</td><td>non-ionic contrast medium for DECT scan</td></tr><tr><td>normal saline</td><td>HUNAN KELUN PHARMACEUTICAL CO,LTD</td><td>NMPN:H43020455</td><td>placebo of control group</td></tr><tr><td>BALB/c nude mice&amp;nbsp;</td><td>SLAC LABORATORY ANIMAL</td><td>BALB/cASlac-nu</td><td>animal model</td></tr><tr><td>SGC-7901&amp;nbsp; cells</td><td>Library of typical culture of Chinese academy of sciences</td><td>TCHu 46</td><td>gastric cancer cell&amp;nbsp;</td></tr><tr><td>SB431542</td><td>Selleck</td><td>No.S1067</td><td>TGF-&amp;beta;1 inhibitor</td></tr><tr><td>GE Discovery CT750 HD</td><td>GE Healthcare</td><td></td><td>dual-energy spectral CT scanner&amp;nbsp;</td></tr><tr><td>AW Volumeshare5</td><td>GE Healthcare</td><td></td><td>dual-energy spectral CT workstation</td></tr><tr><td>Siemens Inveon micro-PET/CT</td><td>Siemens Preclinical Solution</td><td></td><td>positron emission tomography/&lt;br/&gt; computed tomography scanner&amp;nbsp;</td></tr><tr><td>Inveon Acquisition Workplace</td><td>Siemens Preclinical Solution</td><td></td><td>PET-CT workstation</td></tr></tbody></table>

gene regulation and targeted therapy in gastric cancer peritoneal metastasis: radiological findings from dual energy ct and pet/ct

胃癌在全世界的癌症发病率中仍然是第四, 五年的存活率只有 20%-30%。腹膜转移是胃癌最常见的转移类型, 是预后的决定性因素。预防和控制腹膜转移的发展, 有助于延长胃癌患者的生存期。非侵入性和高效的影像学技术将帮助我们识别腹膜转移的侵袭和转移过程, 并监测肿瘤结节的变化, 以应对治疗。这将使我们能够准确地描述胃癌的发展过程和分子机制。我们最近用双能量 CT (DECT) 和正电子发射断层扫描/计算机断层扫描 (PET/ct) 平台进行了实验, 以检测和监测裸鼠胃肿瘤转移的模型。我们已经表明, 每周连续监测与 DECT 和 PET/CT 可以识别动态变化的腹膜转移。sFRP1-overexpression 在胃癌模型中的作用表现为阳性的放射性表现, 较高的葡萄糖摄取和增强, 而 SUV 的max (标准摄取值) 显示出明显的改变趋势对 TGF-β1抑制剂靶向治疗的反应。在本文中, 我们描述了详细的非侵入性成像程序, 以进行更复杂的研究胃癌腹膜转移使用动物模型, 并提供了代表性的成像结果。利用非侵入性成像技术, 我们应该能够更好地了解肿瘤发生的机制, 监测癌症的生长, 并评估治疗干预对胃癌的影响。

在两周的细胞线注射后, 对裸鼠进行了 DECT 和 PET/CT 扫描。影像学图像在 sFRP1 过度表达组的腹部轮廓外显示皮下转移, 并通过彩色比例图像 (图 1-c) 进行外周增强的转移, 取得了良好的效果。PET/CT 图像描述局部异常葡萄糖摄取转移, 包括在腹膜和皮下转移 (图 1d)。DECT 和 PET/CT 图像上显示的腹膜转移和大皮下转移进一步说明了大体标本和组织学切片 (图 1e-f)。与阳性表达组相比, 在 sFRP1 空载组的腹腔内, DECT 和 PET/CT 图像 (图 2a-b) 没有明显的病灶, 显著的增强, 或高的葡萄糖摄取。虽然大体标本和组织学结果的图像证实了该组的成功植入 (图 2c-d)。
TGF-β1抑制剂和 placebowere 对裸鼠进行两周细胞线注射后的干预治疗, 两周后裸鼠进行 DECT 和 PET CT 扫描。为确定从靶向治疗开始的小鼠腹膜转移结节的形成过程, 进行了随访 DECT 和 PET/CT 扫描。进行非侵入性影像扫描, 以评估 TGF-β1靶向治疗的效果。TGF-β1治疗组的小鼠图像显示, DECT 和 PET/CT (图 3a-b) 的冠状融合图像中明显增强和焦点异常的葡萄糖摄取转移。毛标本只说明腹膜转移灶的8结节 (图 3c), 腹腔扩散分布。定量地,图 3显示了 DECT 的适度外围增强和降低的葡萄糖摄取量, 而 SUVmax接近0.83。另一方面, 对照组正常生理盐水的小鼠在 DECT 和 PET/CT (图 4a-b) 的冠状融合图像中也显示出明显的病灶和局部异常摄取转移。总标本说明了22例腹膜转移瘤 (图 4c), 局部转移结节附着于腹腔。对照组小鼠的肿瘤 SUVmax 值未发生改变 (1.26)。
值得注意的是, 有时肠道会导致葡萄糖轻度摄取和图像中的明亮区域会产生假阳性结果。心脏和膀胱也会聚集大量的葡萄糖, 这可能显示为一个亮点的图像。有必要避免相关的水平图像, 以确定真正的葡萄糖摄取面积的肿瘤。
<img alt="Figure 1" class="xfigimg" src="/files/ftp_upload/56526/56526fig1.jpg"> 图 1: 图 sFRP1 过度表达组腹膜转移模型的 DECT、PET/CT 及相应 he 染色.(a-c)在门户阶段的单色图像: (a) 横向单色图像, (b) 横向彩色融合图像, (c) 日冕色标图像;(d) PET/CT 的冠状融合图像 (e) 总标本 (f) 组织学切片。箭头表明腹膜转移结节, 而箭头表明皮下转移。图 1f是肿瘤结节的病理染色结果;图像显示肿瘤细胞的形态和分布;缩放条 = 100 µm。此图已从引用19中进行了修改。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56526/56526fig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 2" class="xfigimg" src="/files/ftp_upload/56526/56526fig2.jpg"> 图 2: 图 sFRP1 空负荷组腹膜转移模型的 DECT、PET/CT 及组织学分析.(a) 在门阶段获得的彩色比例融合图像, (b) 融合 PET/CT 图像, (c) 总标本, 和 (d) 组织学部分。腹腔内无可见病灶, 明显异常增强或高葡萄糖摄取。(c) 和 (d) 确认成功植入。箭头在图 2c中指出了由 SGC-7901/vector 细胞系引起的腹膜转移结节。心脏和膀胱在图 2b中显示了明显的葡萄糖升高。图 2d是肿瘤结节的病理染色结果;图像显示肿瘤细胞的形态和分布;缩放条 = 100 µm。此图已从引用19中进行了修改。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56526/56526fig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 3" class="xfigimg" src="/files/ftp_upload/56526/56526fig3.jpg"> 图 3: 图 TGF-β1治疗组腹膜转移模型的 DECT 和 PET/CT 及相应的大体标本.(a) DECT 的冠状融合图像 (b) PET/CT 的融合图像. (c) 总标本。心脏和膀胱显示明显的葡萄糖海拔在图 3b。总标本说明8例腹膜转移瘤。箭头指出了与影像学发现相对应的转移性结节。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56526/56526fig3large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Figure 4" class="xfigimg" src="/files/ftp_upload/56526/56526fig4.jpg"> 图 4: 图 TGF-β1对照组腹膜转移模型的 DECT 和 PET/CT 及相应的大体标本.(a) DECT 的冠状融合图像 (b) PET/CT 的熔融图像, 以及 (c) 总标本。总标本说明22例腹膜转移瘤。箭头指出了与影像学发现相对应的转移性结节。<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56526/56526fig4large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Supplementary Figure 1" src="/files/ftp_upload/56526/56526supfig1.jpg"> 补充图 1: 样本横向颜色融合图像.<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56526/56526supfig1large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>
<img alt="Supplementary Figure 2" src="/files/ftp_upload/56526/56526supfig2.jpg"> 补充图 2: 样品冠状颜色融合图像.<a href="//cloudflare-jove-com.remotexs.ntu.edu.sg/files/ftp_upload/56526/56526supfig2large.jpg" target="_blank">请单击此处查看此图的较大版本.</a>

Watch this Scientific Journal Video about Gene Regulation and Targeted Therapy in Gastric Cancer Peritoneal Metastasis: Radiological Findings from Dual Energy CT and PET/CT at JoVE.com

Gene Regulation and Targeted Therapy in Gastric Cancer Peritoneal Metastasis: Radiological Findings from Dual Energy CT and PET/CT

本协议描述了双能量 ct 和 PET/ct 成像方法在肿瘤成像和疗效评价中的价值。本文阐述了双能量 ct 和 PET/ct 对胃癌腹膜转移的基因调控和靶向治疗的研究方法和结果。

胃癌腹膜转移的基因调控及靶向治疗: 双能量 ct 与 PET/ct 的影像学研究

Gastric cancer remains fourth in cancer incidence worldwide with a five-year survival of only 20%-30%. Peritoneal metastasis is the most frequent type of ...

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10.8K 次观看.  Shanghai Jiao Tong University School of Medicine. 本协议描述了双能量 ct 和 PET/ct 成像方法在肿瘤成像和疗效评价中的价值。本文阐述了双能量 ct 和 PET/ct 对胃癌腹膜转移的基因调控和靶向治疗的研究方法和结果。

视频: Gene Regulation and Targeted Therapy in Gastric Cancer Peritoneal Metastasis: Radiological Findings from Dual Energy CT and PET/CT

Looking for Driver Pathways of Acquired Resistance to Targeted Therapy: Drug Resistant Subclone Generation and Sensitivity Restoring by Gene Knock-down

The past two decades have seen a shift from cytotoxic drugs to targeted therapy in medical oncology. Although targeted therapeutic agents have shown more impressive clinical efficacy and minimized adverse effects than traditional treatments, drug resistance has become the main limitation to their benefits. Several preclinical in vitro/in vivo models of acquired resistance to targeted agents in clinical practice have been developed mainly by using two strategies: i) genetic manipulation for modeling genotypes of acquired resistance, and ii) in vitro/in vivo selection of resistant models. In the present work, we propose a unifying framework, for investigating the underlying mechanisms responsible for acquired resistance to targeted therapeutic agents, starting from the generation of drug-resistant cellular subclones to the description of silencing procedures used for restoring the sensitivity to the inhibitor. This simple time- and cost-effective approach is widely applicable, and could be easily extended to investigate resistance mechanisms to other targeted therapeutic drugs in different tumor histotypes.

This is a time- and cost-effective in vitro protocol investigating the mechanisms of acquired resistance to targeted therapeutic agents, which is a highly unmet medical need in cancer management.

寻找获得性抗靶治疗的驱动途径: 耐药亚克隆生成与基因击倒敏感性恢复

The past two decades have seen a shift from cytotoxic drugs to targeted therapy in medical oncology. Although targeted therapeutic agents have shown more ...

科研

癌症研究

PET and MRI Guided Irradiation of a Glioblastoma Rat Model Using a Micro-irradiator

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without the ability to target a specific or well-delineated tumor volume. The delivery of radiation was achieved using fixed radiation sources or linear accelerators producing megavoltage (MV) X-rays. These devices are unable to achieve sub-millimeter precision required for small animals. Furthermore, the high doses delivered to healthy surrounding tissue hamper response assessment. To increase the translation between small animal studies and humans, our goal was to mimic the treatment of human glioblastoma in a rat model. To enable a more accurate irradiation in a preclinical setting, recently, precision image-guided small animal radiation research platforms were developed. Similar to human planning systems, treatment planning on these micro-irradiators is based on computed tomography (CT). However, low soft-tissue contrast on CT makes it very challenging to localize targets in certain tissues, such as the brain. Therefore, incorporating magnetic resonance imaging (MRI), which has excellent soft-tissue contrast compared to CT, would enable a more precise delineation of the target for irradiation. In the last decade also biological imaging techniques, such as positron emission tomography (PET) gained interest for radiation therapy treatment guidance. PET enables the visualization of e.g., glucose consumption, amino-acid transport, or hypoxia, present in the tumor. Targeting those highly proliferative or radio-resistant parts of the tumor with a higher dose could give a survival benefit. This hypothesis led to the introduction of the biological tumor volume (BTV), besides the conventional gross target volume (GTV), clinical target volume (CTV), and planned target volume (PTV).
At the preclinical imaging lab of Ghent University, a micro-irradiator, a small animal PET, and a 7 T small animal MRI are available. The goal was to incorporate MRI-guided irradiation and PET-guided sub-volume boosting in a glioblastoma rat model.

In the past, small animal irradiation was usually performed without the ability to target a well-delineated tumor volume. The goal was to mimic the treatment of human glioblastoma in rats. Using a small animal irradiation platform, we performed MRI-guided 3D conformal irradiation with PET-based sub-volume boosting in a preclinical setting.

PET 与 MRI 引导下微器对大鼠胶质瘤模型的照射

For decades, small animal radiation research was mostly performed using fairly crude experimental setups applying simple single-beam techniques without ...

Utilizing 18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in Mouse Models of Lung Cancer

A hallmark of advanced tumors is a switch to aerobic glycolysis that is readily measured by [18F]-2-fluoro-2-deoxy-D-glucose positron emission tomography (18F-FDG PET) imaging. Co-mutations in the KRAS proto-oncogene and the LKB1 tumor suppressor gene are frequent events in lung cancer that drive hypermetabolic, glycolytic tumor growth. A critical pathway regulating the growth and metabolism of these tumors is the mechanistic target of the rapamycin (mTOR) pathway, which can be effectively targeted using selective catalytic mTOR kinase inhibitors. The mTOR inhibitor MLN0128 suppresses glycolysis in mice bearing tumors with Kras and Lkb1 co-mutations, referred to as KL mice. The therapy response in KL mice is first measured by 18F-FDG PET and computed tomography (CT) imaging before and after the delivery of MLN0128. By utilizing 18F-FDG PET/CT, researchers are able to measure dynamic changes in the glucose metabolism in genetically engineered mouse models (GEMMs) of lung cancer following a therapeutic intervention with targeted therapies. This is followed by ex vivo autoradiography and a quantitative immunohistochemical (qIHC) analysis using morphometric software. The use of qIHC enables the detection and quantification of distinct changes in the biomarker profiles following treatment as well as the characterization of distinct tumor pathologies. The coupling of PET imaging to quantitative histology is an effective strategy to identify metabolic and therapeutic responses in vivo in mouse models of disease.

Utilizing 18F-FDG PET/CT Imaging and Quantitative Histology to Measure Dynamic Changes in the Glucose Metabolism in Mouse Models of Lung Cancer

In this protocol, we describe how to utilize [18F]-2-fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography (18F-FDG PET/CT) imaging to measure the tumor metabolic response to the targeted therapy MLN0128 in a Kras/Lkb1 mutant mouse model of lung cancer and coupled imaging with high resolution ex vivo autoradiography and quantitative histology.

利用18f-葡萄糖 PET/CT 成像和定量组织学测量肺癌小鼠模型葡萄糖代谢的动态变化

A hallmark of advanced tumors is a switch to aerobic glycolysis that is readily measured by [18F]-2-fluoro-2-deoxy-D-glucose positron emission tomography ...

A Practical Guide for the Production and PET/CT Imaging of 68Ga-DOTATATE for Neuroendocrine Tumors in Daily Clinical Practice

Neuroendocrine tumors are a rare form of cancer that arise from neuroendocrine cells and can be present at almost any location throughout the body. Although heterogeneous in presentation, a common denominator among these tumors is the overexpression of somatostatin receptors. 68Ga-DOTATATE is a somatostatin analog labeled with the positron emitter gallium-68 (68Ga). For well-differentiated neuroendocrine tumors, 68Ga-DOTATATE positron emission tomography (PET)/computed tomography (CT) imaging is used for diagnosis, determination of disease burden, and therapy selection.
This protocol details the radiolabeling of 68Ga-DOTATATE, quality control, patient preparation, and subsequent PET/CT imaging. Radiolabeling of 68Ga-DOTATATE is performed with a fully automated labeling module coupled to a germanium-68 (68Ge)/68Ga generator. Quality control of the final product evaluates radiochemical purity with instant thin-layer chromatography and solid-phase chromatography, and pH prior to patient injection. Periodic quality control is performed to determine 68Ge breakthrough, sterility, and (4-(2-hydroxyethyl)-1-piperazineethanesulfonic acid (HEPES) content. Patient preparation includes patient instructions, a protocol for 68Ga-DOTATATE during treatment with somatostatin analogs, and intravenous administration of the radiopharmaceutical. For PET/CT imaging, the acquisition and reconstruction settings are described. For each step, radiation safety will be highlighted, as well as time constrictions due to the short half-life of 68Ga.
Fully automated in-house production and quality control of 68Ga-DOTATATE leads to very high success rates (95%) and produces two to four patient dosages per batch, depending on the yield of the generator. In conclusion, 68Ga-DOTATATE PET/CT imaging is a noninvasive and fast method of providing information on the tumor burden of neuroendocrine tumors (NETs) while also assisting in diagnosis and therapy selection.

A Practical Guide for the Production and PET/CT Imaging of 68Ga-DOTATATE for Neuroendocrine Tumors in Daily Clinical Practice

Well-differentiated neuroendocrine tumors overexpress somatostatin receptors which can be utilized for diagnostic imaging with the radiolabeled somatostatin analog 68Ga-DOTATATE. This protocol details the radiolabeling of 68Ga-DOTATATE, quality control, patient preparation, and subsequent PET/CT imaging. Radiation safety and time constrictions due to the short half-life of 68Ga are taken into account.

68型 Ga-DOTATATE 神经内分泌肿瘤的日常临床应用指南

Neuroendocrine tumors are a rare form of cancer that arise from neuroendocrine cells and can be present at almost any location throughout the body. ...

Testing Targeted Therapies in Cancer using Structural DNA Alteration Analysis and Patient-Derived Xenografts

We present here an integrative approach for testing efficacy of targeted therapies that combines the next generation sequencing technolo-gies, therapeutic target analyses and drug response monitoring using patient derived xenografts (PDX). This strategy was validated using ovarian tumors as an example. The mate-pair next generation sequencing (MPseq) protocol was used to identify structural alterations and followed by analysis of potentially targetable alterations. Human tumors grown in immunocompromised mice were treated with drugs selected based on the genomic analyses. Results demonstrated a good correlation between the predicted and the observed responses in the PDX model. The presented approach can be used to test the efficacy of combination treatments and aid personalized treatment for patients with recurrent cancer, specifically in cases when standard therapy fails and there is a need to use drugs off label.

Here we present a protocol to test the efficacy of targeted therapies selected based on the genomic makeup of a tumor. The protocol describes identification and validation of structural DNA rearrangements, engraftment of patients&#8217; tumors into mice and testing responses to corresponding drugs.

使用结构DNA改变分析和患者衍生异种移植测试癌症的靶向疗法

We present here an integrative approach for testing efficacy of targeted therapies that combines the next generation sequencing technolo-gies, therapeutic ...

Dynamic Contrast Enhanced Magnetic Resonance Imaging of an Orthotopic Pancreatic Cancer Mouse Model

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe respiratory motion artifact in the abdominal area. Orthotopic tumor models offer advantages over subcutaneous ones, because those can reflect the primary tumor microenvironment affecting blood supply, neovascularization, and tumor cell invasion. We have recently established a protocol of DCE-MRI of orthotopic pancreatic tumor xenografts in mouse models by securing tumors with an orthogonally bent plastic board to prevent motion transfer from the chest region during imaging. The pressure by this board was localized on the abdominal area, and has not resulted in respiratory difficulty of the animals. This article demonstrates the detailed procedure of orthotopic pancreatic tumor modeling using small animals and DCE-MRI of the tumor xenografts. Quantification method of pharmacokinetic parameters in DCE-MRI is also introduced. The procedure described in this article will assist investigators to apply DCE-MRI for orthotopic gastrointestinal cancer mouse models.

The goal of this protocol is to apply dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) for orthotopic pancreatic tumor xenografts in mice. DCE-MRI is a non-invasive method to analyze microvasculature in a target tissue, and useful to assess vascular response in a tumor following a novel therapy.

原位胰腺癌小鼠模型的动态对比增强磁共振成像

Dynamic contrast enhanced magnetic resonance imaging (DCE-MRI) has been limitedly used for orthotopic pancreatic tumor xenografts due to severe ...

医学

Quantitation of Intra-peritoneal Ovarian Cancer Metastasis

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States. Mortality is due to diagnosis of 75% of women with late stage disease, when metastasis is already present. EOC is characterized by diffuse and widely disseminated intra-peritoneal metastasis. Cells shed from the primary tumor anchor in the mesothelium that lines the peritoneal cavity as well as in the omentum, resulting in multi-focal metastasis, often in the presence of peritoneal ascites. Efforts in our laboratory are directed at a more detailed understanding of factors that regulate EOC metastatic success. However, quantifying metastatic tumor burden represents a significant technical challenge due to the large number, small size and broad distribution of lesions throughout the peritoneum. Herein we describe a method for analysis of EOC metastasis using cells labeled with red fluorescent protein (RFP) coupled with in vivo multispectral imaging. Following intra-peritoneal injection of RFP-labelled tumor cells, mice are imaged weekly until time of sacrifice. At this time, the peritoneal cavity is surgically exposed and organs are imaged in situ. Dissected organs are then placed on a labeled transparent template and imaged ex vivo. Removal of tissue auto-fluorescence during image processing using multispectral unmixing enables accurate quantitation of relative tumor burden. This method has utility in a variety of applications including therapeutic studies to evaluate compounds that may inhibit metastasis and thereby improve overall survival.

Ovarian cancer metastasis is characterized by numerous diffuse intra-peritoneal lesions, such that accurate visual quantitation of tumor burden is challenging. Herein we describe a method for in situ and ex vivo quantitation of metastatic tumor burden using red fluorescent protein (RFP)-labeled tumor cells and optical imaging.

腹膜内卵巢癌转移的定量

Epithelial ovarian cancer (EOC) is the leading cause of death from gynecologic malignancy in the United States.  Mortality is due to diagnosis of 75% of ...

Implantation and Monitoring by PET/CT of an Orthotopic Model of Human Pleural Mesothelioma in Athymic Mice

Malignant pleural mesothelioma (MPM) is a rare and aggressive tumor arising in the mesothelium that covers the lungs, the heart, and the thoracic cavity. MPM development is mainly associated with asbestos. Treatments provide only modest survival since the median survival average is 9&#8211;18 months from the time of diagnosis. Therefore, more effective treatments must be identified. Most data describing new therapeutic targets were obtained from in vitro experiments and need to be validated in reliable in vivo preclinical models. This article describes one such reliable MPM orthotopic model obtained after injection of a human MPM cell line H2052/484 into the pleural cavity of immunodeficient athymic mice. Transplantation in the orthotopic site allows studying the progression of tumor in the natural in vivo environment. Positron emission tomography/computed tomography (PET/CT) molecular imaging using the clinical [18F]-2-fluoro-2-deoxy-D-glucose ([18F]FDG) radiotracer is the diagnosis method of choice for examining patients with MPM. Accordingly, [18F]FDG-PET/CT was used to longitudinally monitor the disease progression of the H2052/484 orthotopic model. This technique has a high 3R potential (Reduce the number of animals, Refine to lessen pain and discomfort, and Replace animal experimentation with alternatives) since the tumor development can be monitored non-invasively and the number of animals required could be significantly reduced.
This model displays a high development rate, a rapid tumor growth, is cost-efficient and allows for rapid clinical translation. By using this orthotopic xenograft MPM model, researchers can assess biological responses of a reliable MPM model following therapeutic interventions.

This article describes the generation of an orthotopic mouse model of human pleural mesothelioma by implantation of H2052/484 mesothelioma cells into the pleural cavity of immunocompromised athymic mice. The longitudinal monitoring of the development of intrapleural tumors was assessed by non-invasive multimodal [18F]-2-fluoro-2-deoxy-D-glucose positron emission tomography and computed tomography imaging.

小鼠中人类胸膜皮质瘤的PET/CT的植入与监测

Malignant pleural mesothelioma (MPM) is a rare and aggressive tumor arising in the mesothelium that covers the lungs, the heart, and the thoracic cavity. ...

免疫与感染

A Preclinical Human-Derived Autologous Gastric Cancer Organoid/Immune Cell Co-Culture Model to Predict the Efficacy of Targeted Therapies

Tumors expressing programmed cell death-ligand 1 (PD-L1) interact with programmed cell death protein 1 (PD-1) on CD8+ cytotoxic T lymphocytes (CTLs) to evade immune surveillance leading to the inhibition of CTL proliferation, survival, and effector function, and subsequently cancer persistence. Approximately 40% of gastric cancers express PD-L1, yet the response rate to immunotherapy is only 30%. We present the use of human-derived autologous gastric cancer organoid/immune cell co-culture as a preclinical model that may predict the efficacy of targeted therapies to improve the outcome of cancer patients. Although cancer organoid co-cultures with immune cells have been reported, this co-culture approach uses tumor antigen to pulse the antigen-presenting dendritic cells. Dendritic cells (DCs) are then cultured with the patient's CD8+ T cells to expand the cytolytic activity and proliferation of these T lymphocytes before co-culture. In addition, the differentiation and immunosuppressive function of myeloid-derived suppressor cells (MDSCs) in culture are investigated within this co-culture system. This organoid approach may be of broad interest and appropriate to predict the efficacy of therapy and patient outcome in other cancers, including pancreatic cancer.

The goals of the protocol are to use this approach to 1) understand the role of the immunosuppressive gastric tumor microenvironment and 2) predict the efficacy of patient response, thus increasing the survival rate of patients.

Tumors expressing programmed cell death-ligand 1 (PD-L1) interact with programmed cell death protein 1 (PD-1) on CD8+ cytotoxic T lymphocytes (CTLs) to ...

使用 Ion Torrent 测序的 SNP 指导的胃癌 5-FU 疗法

Multi-Gene Single Nucleotide Polymorphism Detection in Gastric Cancer Based on Ion Semiconductor Sequencing Platform

Gastric cancer is a common heterogeneous tumor. Most patients have advanced gastric cancer at the time of diagnosis and often need chemotherapy. Although 5-fluorouracil (5-FU) is widely used for treatment, its therapeutic sensitivity and drug tolerance still need to be determined, which emphasizes the importance of individualized administration. Pharmacogenetics can guide the clinical implementation of individualized treatment. Single nucleotide polymorphisms (SNPs), as a genetic marker, contribute to the selection of appropriate chemotherapy regimens and dosages. Some SNPs are associated with folate metabolism, the therapeutic target of 5-FU. Methylenetetrahydrofolate reductase (MTHFR) rs1801131 and rs1801133, dihydrofolate reductase (DHFR) rs1650697 and rs442767, methionine synthase (MTR) rs1805087, gamma-glutamyl hydrolase (GGH) rs11545078 and solute carrier family 19 member 1 (SLC19A1) rs1051298 have been investigated in different kinds of cancers and antifolate antitumor drugs, which have potential forecasting and guiding significance for application of 5-FU. The ion torrent next-generation semiconductor sequencing technology can rapidly detect gastric cancer-related SNPs. Each time a base is extended in a DNA chain, an H+ will be released, causing local pH changes. The ionic sensor detects pH changes and converts chemical signals into digital signals, achieving sequencing by synthesis. This technique has low sample requirement, simple operation, low cost, and fast sequencing speed, which is beneficial for guiding individualized chemotherapy by SNPs.

作者聚焦：胃癌氟尿嘧啶反应的遗传分析

This protocol proposes holistic laboratory procedures required to detect single nucleotide polymorphisms in gastric cancer samples based on an ion semiconductor sequencing platform. The target sequences, ligating adapters, library amplification and purification, and quality control criteria are also described in detail.

基于离子半导体测序平台的胃癌多基因单核苷酸多态性检测

Gastric cancer is a common heterogeneous tumor. Most patients have advanced gastric cancer at the time of diagnosis and often need chemotherapy. Although ...

胃癌腹膜转移的基因调控及靶向治疗: 双能量 ct 与 PET/ct 的影像学研究

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