Tsuji Kazuhiko, Yamauchi Kensuke, Yang Meng, Jiang Ping, Bouvet Michael, Endo Hitoshi, Kanai Yoshikatsu, Yamashita Koji, Moossa Abdool R, Hoffman Robert M
AntiCancer, Inc., San Diego, California 92111, USA.
Cancer Res. 2006 Jan 1;66(1):303-6. doi: 10.1158/0008-5472.CAN-05-2958.
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-116-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.
我们使用双色体内细胞成像技术来观察癌细胞注射到小鼠门静脉后的运输、核质动力学以及细胞活力。在这些研究中，我们使用了双色荧光癌细胞，其细胞核中表达与组蛋白H2B相连的绿色荧光蛋白（GFP），细胞质中表达逆转录病毒红色荧光蛋白（RFP）。将人HCT-116-GFP-RFP结肠癌细胞和小鼠乳腺肿瘤（MMT）细胞注射到裸鼠的门静脉中。使用奥林巴斯OV100全小鼠成像系统在单细胞水平对肝脏中的细胞进行活体观察。大多数HCT-116-GFP-RFP细胞留在外周门静脉附近的肝血窦中。只有一小部分癌细胞侵入小叶区域。HCT-116-GFP-RFP细胞在6小时内发生广泛的胞质溶解（细胞质破坏）。注射后12小时内门静脉内凋亡细胞数量迅速增加。通过双色细胞改变的核形态很容易观察到凋亡。数据表明HCT-116-GFP-RFP细胞在门静脉中迅速死亡。相比之下，注射到门静脉中的双色MMT-GFP-RFP细胞在注射后24小时大多在裸鼠肝脏中存活。许多存活的MMT-GFP-RFP细胞显示出带有细胞质突起的侵袭性形态。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能在肝脏中存活并形成集落。这些结果表明，对环磷酰胺敏感的宿主细胞系统攻击了HCT-116-GFP-RFP细胞，但不能有效地杀死MMT-GFP-RFP细胞。
