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稳定表达萤火虫荧光素酶的Hepa1-6-FLuc细胞系保留了其原始特性,并具有良好的生物发光成像能力。

Hepa1-6-FLuc cell line with the stable expression of firefly luciferase retains its primary properties with promising bioluminescence imaging ability.

作者信息

Li Yasha, Liu Mengnan, Cui Jiejie, Yang Ke, Zhao Li, Gong Mengjia, Wang Yi, He Yun, He Tongchuan, Bi Yang

机构信息

Stem Cell Biology and Therapy Laboratory, Ministry of Education Key Laboratory of Child Development and Disorders, The Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.

Department of Pediatric Surgery, The Children's Hospital of Chongqing Medical University, Chongqing 400014, P.R. China.

出版信息

Oncol Lett. 2018 May;15(5):6203-6210. doi: 10.3892/ol.2018.8132. Epub 2018 Feb 27.

Abstract

Reliable animal models are required for the study of the molecular mechanisms and effects of chemotherapeutic drugs in hepatocarcinoma. tracing techniques based on firefly luciferase (FLuc) may optimize the non-invasive monitoring of experimental animals. The present study established a murine Hepa1-6-FLuc cell line that stably expressed a retrovirus-delivered FLuc protein gene. The cell morphology, proliferation, migration and invasion ability of Hepa1-6-FLuc cells were the same as that of the Hepa1-6 cells, and thus is suitable to replace Hepa1-6 cells in the construction of hepatocarcinoma animal models. No differences in subcutaneous tumor mass and its pathomorphology from implanted Hepa1-6-FLuc cells were observed compared with Hepa1-6 control tumors. Bioluminescence imaging indicated that the Luc signal of the Hepa1-6-FLuc cells was consistently strengthened with increases in tumor mass; however, the Luc signal of Hepa1-6-AdFLuc became weaker and eventually disappeared during tumor development. Therefore, compared with the transient expression by adenovirus, stable expression of the FLuc gene in Hepa1-6 cells may better reflect cell proliferation and survival , and provide a reliable source for the establishment of hepatocarcinoma models.

摘要

肝癌化疗药物分子机制及效应研究需要可靠的动物模型。基于萤火虫荧光素酶(FLuc)的示踪技术可优化对实验动物的无创监测。本研究建立了稳定表达逆转录病毒传递的FLuc蛋白基因的小鼠Hepa1-6-FLuc细胞系。Hepa1-6-FLuc细胞的细胞形态、增殖、迁移和侵袭能力与Hepa1-6细胞相同,因此适合在肝癌动物模型构建中替代Hepa1-6细胞。与Hepa1-6对照肿瘤相比,未观察到植入Hepa1-6-FLuc细胞后皮下肿瘤块及其病理形态有差异。生物发光成像表明,Hepa1-6-FLuc细胞的Luc信号随肿瘤块增大而持续增强;然而,Hepa1-6-AdFLuc的Luc信号在肿瘤发展过程中变弱并最终消失。因此,与腺病毒的瞬时表达相比,FLuc基因在Hepa1-6细胞中的稳定表达可能更好地反映细胞增殖和存活情况,并为建立肝癌模型提供可靠来源。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/5581/5876459/2e2879556d75/ol-15-05-6203-g00.jpg

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