建立分泌型荧光素酶基因标记的原位肝癌模型及用于干扰素β基因治疗评价

Application of secretary luciferase labeled orthotopic transplant model of hepatocellular carcinoma to evaluate tumor response to interferon-β gene therapy

摘要:

旨在建立一种分泌型荧光素酶基因标记的小鼠原位移植型肝癌模型并观察其对干扰素β基因治疗的反应。首先建立稳定表达分泌型荧光素酶Gluc (Gaussia princeps luciferase) 的小鼠肝癌细胞Hepa 1-6/Gluc;将该细胞通过脾注射至C57BL/6小鼠肝脏建立原位移植型肝癌模型，通过检测外周血Gluc活性监测小鼠体内肿瘤生长情况；用此模型观察水动力注射干扰素β质粒DNA的抗肿瘤效果。结果表明，通过脾注射Gluc基因标记的Hepa 1-6细胞可以建立小鼠原位移植型肝癌模型;外周血Gluc活性可以有效反映体内接种肿瘤细胞的数量和肿瘤的生长情况;通过监测外周血Gluc活性可灵敏反映干扰素β基因治疗对肿瘤生长的抑制作用。本研究表明，利用Gluc为报告基因建立的小鼠原位移植型肝癌模型可以体外实时监测肿瘤的生长情况，并能灵敏可靠地用于抗肿瘤治疗效果的评价。

Abstract:

To establish an orthotopic transplant mouse model of hepatocellular carcinoma (HCC) labeled with secretary luciferase and to study its response to anti-tumor treatment with interferon-β gene therapy. We labeled the murine hepatoma Hepa1-6 cells with secretary Gaussia princeps luciferase (Gluc), and then injected Gluc labeled Hepa1-6 cells intrasplenically in C57BL/6 mice. We monitored blood Gluc to evaluate the tumor development and anti-tumor effects of hydrodynamic injection with interferon-β expressing plasmid. We successfully established the orthotopic mouse model of HCC by intrasplenic injection of Gluc labeled Hepa1-6 cells. The Gluc blood assay could reflect the amount of cancer cells in vivo, tumor progression, as well as anti-tumor effect of interferon-β gene therapy. In conclusion, Gluc labeled orthotopic transplant mouse model of HCC can ex vivo real-time monitor the tumor development and tumor response to treatments.