GFP/Luc双标食管癌细胞动物模型建立及其活体成像观察
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摘要
目的建立绿色荧光蛋白(GFP)/荧光素酶(Luc)双标的人食管癌ECA109细胞系(ECA109-Luc-GFP),采用一种非侵入性的生物活体成像方法监测食管癌裸鼠肿瘤模型。方法 MTT、流式细胞仪、划痕实验分析法检测ECA109-Luc-GFP细胞系与正常ECA109细胞的生物学特性差异。利用活体成像系统示踪肿瘤。结果与正常ECA109相比,ECA109-Luc-GFP细胞在细胞活力、增殖和迁移能力方面差异均无显著性(P> 0.05)。利用小动物活体成像实验技术观察了肿瘤生长、转移等生物学特性。结论成功构建了GFP/Luc双标的人食管癌ECA109细胞系(ECA109-Luc-GFP),应用小动物活体成像应用技术并活体观察食管癌动物模型的生长、转移。
Objective To establish a green fluorescent protein(GFP)/firefly luciferase(Luc) double-labeled ECA109 human esophageal cancer cells(ECA109-Luc-GFP) and apply them to a mouse model of esophageal cancer for non-invasive bioimaging. Methods MTT, flow cytometry(FCM), and wound healing assays were used to analyze biological differences between ECA109-Luc-GFP cells and parental ECA109 cells. Esophageal cancer xenografts were established by subcutaneous or tail vein injections of ECA109-Luc-GFP cells into nude mice. An in vivo imaging system was used to observe the growth of xenografts. Results MTT, FCM, and wound healing assays showed no differences between ECA109-Luc-GFP and parental cells in terms of cell viability, proliferation, and metastasis(P>0.05). The growth and metastasis of xenografts established by ECA109-Luc-GFP cells were observed using the in vivo imaging system. Conclusions A GFP/Luc double-labeled human esophageal cancer cell line is established, and the growth and metastasis of xenografts can be observed by an in vivo imaging system. This study provides the experimental basis for an esophageal cancer mouse model and clinical study.
Objective To establish a green fluorescent protein(GFP)/firefly luciferase(Luc) double-labeled ECA109 human esophageal cancer cells(ECA109-Luc-GFP) and apply them to a mouse model of esophageal cancer for non-invasive bioimaging. Methods MTT, flow cytometry(FCM), and wound healing assays were used to analyze biological differences between ECA109-Luc-GFP cells and parental ECA109 cells. Esophageal cancer xenografts were established by subcutaneous or tail vein injections of ECA109-Luc-GFP cells into nude mice. An in vivo imaging system was used to observe the growth of xenografts. Results MTT, FCM, and wound healing assays showed no differences between ECA109-Luc-GFP and parental cells in terms of cell viability, proliferation, and metastasis(P>0.05). The growth and metastasis of xenografts established by ECA109-Luc-GFP cells were observed using the in vivo imaging system. Conclusions A GFP/Luc double-labeled human esophageal cancer cell line is established, and the growth and metastasis of xenografts can be observed by an in vivo imaging system. This study provides the experimental basis for an esophageal cancer mouse model and clinical study.
引文
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[2] Ando Y, Niwa K, Yamada N, et al. Firefly bioluminescence quantum yield and colour change by pH-sensitive green emission [J]. Nat Photonics, 2007, 2(1): 44-47.
[3] Tetreault MP. Esophageal cancer: insights from mouse models [J]. Cancer Growth Metastasis, 2015, 8(Suppl 1): 37-46.
[4] Ruijter NCA, Verhees J, Leeuwen W, et al. Evaluation and comparison of the GUS, LUC and GFP reporter system for gene expression studies in plants [J]. Plant Biology, 2003, 5: 103-115.
[5] Saha D, Dunn H, Zhou H, et al. In vivo bioluminescence imaging of tumor hypoxia dynamics of breast cancer brain metastasis in a mouse model [J]. J Vis Exp, 2011, 3(56): 3175.
[6] Jenkins DE, Oei Y, Hornig YS, et al. Bioluminescent imaging (BLI) to improve and refine traditional murine models of tumor growth and metastasis [J]. Clin Exp Metastasis, 2003, 20(8): 733-744.
[7] Doubrovin M, Serganova I, Mayer-Kuckuk P, et al. Multimodality in vivo molecular-genetic imaging [J]. Bioconjug Chem, 2004, 15(6): 1376-1388.
[8] Luker GD, Luker KE. Optical imaging: current applications and future directions [J]. J Nucl Med, 2008, 49(1): 1-4.
[9] Chen ZY, Wang YX, Yang F, et al. New researches and application progress of commonly used optical molecular imaging technology [J]. Biomed Res Int, 2014, 2014: 429198.
[10] Tseng JC, Kung AL. Quantitative bioluminescence imaging of mouse tumor models [J]. Cold Spring Harb Protoc, 2015, 2015(1): pdb.prot078261.
[11] Genevois C, Loiseau H, Couillaud F. In vivo follow-up of brain tumor growth via bioluminescence imaging and fluorescence tomography [J]. Int J Mol Sci, 2016, 17(11): E1815.