为研究雄黄纳米颗粒的体内抗小鼠乳腺癌肺转移和肝转移作用,以萤火虫荧光素酶(luciferase,Luc)基因标记小鼠4T1乳腺癌细胞(4T1-Luc),给BALB/c小鼠尾静脉注射4T1-Luc细胞制作小鼠乳腺癌肺和肝转移瘤模型,纳米雄黄[5mg/(kg·d)和10mg/(kg·d)]灌胃治疗32d,采用小动物活体生物发光成像系统动态观察乳腺癌细胞的转移情况。治疗末期处死动物,取出肺和肝,立即置活体生物发光成像系统下进行成像,然后肉眼观察肺和肝表面转移瘤结节形成并计数;肺、肝组织制片、HE染色,光镜下观察组织形态变化。结果显示,体内纳米雄黄可显著抑制小鼠乳腺癌细胞在肺和肝中的转移(P〈0.05),纳米雄黄治疗组小鼠的肺转移瘤结节和肝微转移灶体积小且数量少。上述结果证实,纳米雄黄可有效抑制小鼠乳腺癌4T1-Luc细胞的肺转移和肝转移能力;活体生物发光成像技术应用于活体肿瘤转移研究较常规动物实验技术具有更高的灵敏性。
To study the anti-pulmonary and-hepatic metastasis actions of realgar nanoparticles(nanorealgar)on breast cancer in mice using the in vivo bioluminescence imaging assay,the firefly luciferase gene was transferred into 4T1 murine breast cancer cells(4T1-Luc).The cells were implanted in female BALB/c mice by tail intravenous injection to establish the pulmonary and hepatic metastasis model of breast cancer.The tumor-metastatic mice were treated with 5mg/(kg·d)and 10 mg/(kg·d)nano-realgar by gavage once a day for 32 days,and the tumor metastatic situation in mice was observed continuously and dynamically using the optical in vivo imaging system.At the end of the treating period,the mice were sacrificed,the lungs and livers were removed and fluorescence images of the organs were acquired immediately under the optical in vivo imaging system and then the metastatic nodules on the lungs and livers surface were observed and counted.The histological sections of lung and liver were stained with HE staining,and the morphological changes of the metastatic tumor were observed under light microscope.The results showed that nano-realgar administration could markedly suppressed the pulmonary and hepatic metastasis ability of4T1-Luc cells in mice(P0.05),and pathological examinations revealed that in the nano-realgar-treated mice,there were much fewer and smaller pulmonary metastasis nodules and hepatic micrometastases than that of in untreated mice.Our data indicate that realgar nanoparticles can significantly weaken the pulmonary and hepatic metastatic abilities of 4T1 murine breast cancer cells,and compared with the conventional animal technique the in vivo bioluminescence imaging exhibits more sensitive in monitoring the tumor metastases in vivo.