Dual-color imaging of nuclear-cytoplasmic dynamics, viability, and proliferation of cancer cells in the portal vein area
Dual-color imaging of nuclear-cytoplasmic dynamics, viability, and proliferation of cancer cells in the portal vein area
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DOI：
10.1158/0008-5472.can-05-2958
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发表时间：
2006-01-01
期刊：
影响因子：
11.2
通讯作者：
Hoffman, RM
中科院分区：
文献类型：
--
作者：
Tsuji, K;Yamauchi, K;Hoffman, RM
我们使用双色体内细胞成像来可视化癌细胞注射到小鼠门静脉后的运输、核质动力学和存活能力。在这些研究中，我们使用了表达与细胞核中的组蛋白H2 B连接的绿色荧光蛋白（GFP）和细胞质中的逆转录病毒红色荧光蛋白（RFP）的双色荧光癌细胞。人HCT-116-GFP-RFP结肠癌细胞和小鼠乳腺肿瘤（MMT）细胞在裸鼠门静脉中为HCT-116-GFP-RFP。使用Olympus OV 100全小鼠成像系统在单细胞水平下在肝脏中活体内观察细胞。大多数HCT-116-GFP-RFP细胞保留在靠近外周门静脉的血窦中。只有一小部分癌细胞侵入小叶区。HCT-116-GFP-RFP细胞的广泛的细胞质破坏（clasmocytosis）在6小时内发生。在注射后12小时内，门静脉内凋亡细胞的数量迅速增加。细胞凋亡是很容易可视化的双色细胞改变核形态。数据表明HCT-116-GFP-RFP细胞在门静脉中快速死亡。相比之下，注射到门静脉中的双色MMT-GFP-RFP细胞在注射后24小时大部分在裸鼠的肝脏中存活。许多存活的MMT-GFP-RFP细胞显示具有细胞质突起的侵袭性图。这些细胞在肝脏中积极生长并形成集落。然而，当宿主小鼠用环磷酰胺预处理时，HCT-116-GFP-RFP细胞在门静脉注射后也存活并在肝脏中形成集落。这些结果表明，环磷酰胺敏感的宿主细胞系统攻击HCT-116-GFP-RFP细胞，但不能有效地杀死MMT-GFP-RFP细胞。
We used dual-color in vivo cellular imaging to visualize trafficking, nuclear-cytoplasmic dynamics, and the viability of cancer cells after their injection into the portal vein of mice. For these studies, we used dual-color fluorescent cancer cells that express green fluorescent protein (GFP) linked to histone H2B in the nucleus and retroviral red fluorescent protein (RFP) in the cytoplasm. Human HCT-116-GFP-RFP colon cancer and mouse mammary tumor (MMT) cells were HCT-116-GFP-RFP in the portal vein of nude mice. The cells were observed intravitally in the liver at the single-cell level using the Olympus OV100 whole-mouse imaging system. Most HCT-116-GFP-RFP cells remained in sinusoids near peripheral portal veins. Only a small fraction of the cancer cells invaded the lobular area. Extensive clasmocytosis (destruction of the cytoplasm) of the HCT-I 16-GFP-RFP cells occurred within 6 hours. The number of apoptotic cells rapidly increased within the portal vein within 12 hours of injection. Apoptosis was readily visualized in the dual-color cells by their altered nuclear morphology. The data suggest rapid death of HCT-116-GFP-RFP cells in the portal vein. In contrast, dual-color MMT-GFP-RFP cells injected into the portal vein mostly survived in the liver of nude mice 24 hours after injection. Many surviving MMT-GFP-RFP cells showed invasive figures with cytoplasmic protrusions. The cells grew aggressively and formed colonies in the liver. However, when the host mice were pretreated with cyclophosphamide, the HCT-116-GFP-RFP cells also survived and formed colonies in the liver after portal vein injection. These results suggest that a cyclophosphamide-sensitive host cellular system attacked the HCT-116-GFP-RFP cells but could not effectively kill the MMT-GFP-RFP cells.