在小鼠模型体内生物发光成像乳腺癌脑转移的肿瘤乏氧动力学
Summary
缺氧诱导因子-1α活动的生物发光成像是用于监测在乳腺癌脑转移瘤小鼠模型的颅内肿瘤缺氧发展。
Abstract
众所周知，肿瘤的缺氧起着促进恶性进展和治疗反应产生负面影响重要作用。有一点知识在原地，在体内 ，在缺氧的发展，因为缺乏有效的手段来监测这些深层次的原位肿瘤恶性脑肿瘤的颅内肿瘤，。生物发光成像（劳保局），表达的荧光素酶基因的活细胞所发出的光检测的基础上，已迅速采用为癌症研究，特别是评估肿瘤的生长或应对，在临床前动物研究治疗肿瘤的大小变化。此外，通过一个控制下的一个子序列的记者基因表达，特定的基因表达是可以被监测的非侵入性劳保局。缺氧压力下，主要通过缺氧诱导因子-1α（HIF -1α）介导的信号反应，以驱动各种基因的转录。因此，我们必须使用一个记者HIF -1α建设，5HRE多- LUC，稳定转染人乳腺癌MDA – MB231 细胞在体外HIF -1α的生物发光检测（MDA-MB231/5HRE-ODD-luc）是由孵化在低氧室，24小时前劳保局（0.1％O 2）转染的细胞，而细胞在常氧（21％O 2）作为一个控制。显着较高的光子通量细胞缺氧条件下的观察表明其启动的增加HIF -1α结合（HRE元素），比在常氧。细胞是直接注射到老鼠大脑建立一个乳腺癌脑转移瘤模型，在体内的肿瘤乏氧动力学生物发光成像。植入后2周开始，并反复每周一次。劳保局揭示增加脑肿瘤进展的光信号，表明增加了颅内肿瘤缺氧。组织学及免疫组织化学研究，以确认在体内成像结果。在这里，我们将介绍在体外 HIF -1α的生物发光检测，乳腺癌裸鼠和应用，在体内的生物发光成像技术来监测颅内肿瘤缺氧的脑转移瘤的手术建立的方法。
Protocol
Discussion
乳腺癌脑转移发生在IV期乳腺癌患者的30％。这是与高发病率和死亡率，并有一个中位生存期13个月6。是需要有合适的动物模型，模仿这种破坏性疾病，在临床上为了便于我们了解其颅内的启动和进展，以及病理生理型材。在这里，我们已经开发注入人类乳腺癌细胞，直接进入鼠脑，原位乳腺癌脑转移模型。我们以前的经验表明，一个放射学可视化（MRI）的颅内病变植入约2周后出现。这个…
Disclosures
The authors have nothing to disclose.
Acknowledgements
这项研究是支持部分由国防部乳腺癌IDEA奖W81XWH – 08 – 1 – 0583和NIH / NCI CA141348 – 01A1（DZ）和FAMRI临床科学家奖（DS）。成像的基础设施是由西南部分小动物成像研究支持计划（U24 CA126608）和西蒙斯癌症中心（P30 CA142543）和NIH 1S10RR024757 – 01。
Materials
| Name of the reagent | Company | Catalogue number | Comments (optional) |
|---|---|---|---|
| D-luciferin | Gold Biotechnology | L-123 | 120 mg/kg in PBS in a total volume of 80 μl for in vivo study |
| Isoflurane | Baxter International Inc. | 1001936060 | |
| Matrigel | BD Biosciences | 354234 | |
| Hamilton syringe | Hamilton Company | 1701 | |
| 32G Hamilton needle | Hamilton Company | 7803-04 | |
| Hypoxia chamber | Billups-Rothenberg, Inc. | MIC-101 | |
| Bioluminescence imaging system | Caliper Life Sciences | IVIS Spectrum system | |
| G418 | Fisher scientific | SV3006901 |
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