Single-photon emission computed tomography of spontaneous liver metastasis from orthotopically implanted human colon cancer cell line stably expressing human sodium/iodide symporter reporter gene.
Single-photon emission computed tomography of spontaneous liver metastasis from orthotopically implanted human colon cancer cell line stably expressing human sodium/iodide symporter reporter gene.
复制标题
单光子发射计算机断层扫描显示稳定表达人钠/碘同向转运蛋白报告基因的原位植入人结肠癌细胞系的自发性肝转移。
DOI：
10.1186/2191-219x-2-46
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发表时间：
2012-09-07
期刊：
影响因子：
3.2
通讯作者：
Saga T
中科院分区：
文献类型：
--
作者：
Inubushi M;Jin YN;Murai C;Hata H;Kitagawa Y;Saga T
我们的目的是利用原位植入的稳定表达人钠/碘同向转运蛋白（NIS）报告基因的人结肠癌细胞系开发小鼠自发性肝转移模型，该模型可以使用 99mTcO4− 通过单光子发射计算机断层扫描（SPECT）进行成像。将含有组成型驱动NIS基因的重组质粒（pcDNA3-NIS）转染人结肠癌细胞系HCT116，建立稳定细胞系。将稳定的细胞皮下注射到裸鼠体内。当直径达到10毫米时，将异种移植物切除，切成小碎片，并原位植入另一只裸鼠的盲肠壁中。 8 周后开始进行 99mTcO4− SPECT/CT 成像，每 1 至 2 周重复一次。 NIS 蛋白的产生和功能通过蛋白质印迹和 99mTcO4− 摄取测定在体外得到证实。在 SPECT/CT 成像中，在肝脏中检测到局部 99mTcO4− 摄取。尸检显示原位结肠异种移植物局部生长，具有广泛的侵袭、显微镜下浆膜转移和相应肝区的转移灶，显示局灶性 99mTcO4− 摄取。免疫组织化学显示形成肝肿瘤的细胞中有高水平的NIS表达，表明肝肿瘤细胞起源于原位结肠异种移植物。目前的概念验证研究为利用放射性核素报告基因对活体小鼠自发性肝转移进行特异性可视化提供了理论依据。这种临床相关且外部可检测的肝转移的独特动物模型将成为研究肿瘤生物学和开发癌症转移新疗法的有力工具。
We aimed to develop a mouse spontaneous liver metastasis model from an orthotopically implanted human colon cancer cell line stably expressing a human sodium/iodide symporter (NIS) reporter gene, which can be imaged with single-photon emission computed tomography (SPECT) using 99mTcO4−. A recombinant plasmid containing a constitutively driven NIS gene (pcDNA3-NIS) was transfected into the human colon cancer cell line HCT116, and stable cell lines were established. The stable cells were subcutaneously injected into the nude mice. When the diameter reached 10 mm, the xenografts were excised, cut into small fragments, and orthotopically implanted into the cecal walls of another nude mice. 99mTcO4− SPECT/CT imaging was initiated 8 weeks later and repeated every 1 to 2 weeks. The production and function of NIS protein was confirmed in vitro by Western blotting and 99mTcO4− uptake assay. On SPECT/CT imaging, focal 99mTcO4− uptake was detected in the liver. Necropsy revealed local growth of the orthotopic colon xenografts with extensive invasion, microscopic serosal metastasis, and metastatic foci in the corresponding hepatic regions showing focal 99mTcO4− uptake. Immunohistochemistry revealed high levels of NIS expression in cells forming liver tumor, indicating that the liver tumor cells originated from the orthotopic colon xenografts. The present proof-of-concept study provided a rationale for employing a radionuclide reporter gene for the specific visualization of spontaneous liver metastasis in living mice. This unique animal model of clinically relevant and externally detectable liver metastasis will be a powerful tool for investigating tumor biology and developing novel therapies for cancer metastasis.