Abstract
Purpose
The development of glioma therapy in clinical practice (e.g., gene therapy) calls for efficiently visualizing and tracking glioma cells in vivo. Human ferritin heavy chain is a novel gene reporter in magnetic resonance imaging. This study proposes hFTH as a reporter gene for MR molecular imaging in glioma xenografts.
Methods
Rat C6 glioma cells were infected by packaged lentivirus carrying hFTH and EGFP genes and obtained by fluorescence-activated cell sorting. The iron-loaded ability was analyzed by the total iron reagent kit. Glioma nude mouse models were established subcutaneously and intracranially. Then, in vivo tumor bioluminescence was performed via the IVIS spectrum imaging system. The MR imaging analysis was analyzed on a 7T animal MRI scanner. Finally, the expression of hFTH was analyzed by western blotting and histological analysis.
Results
Stable glioma cells carrying hFTH and EGFP reporter genes were successfully obtained. The intracellular iron concentration was increased without impairing the cell proliferation rate. Glioma cells overexpressing hFTH showed significantly decreased signal intensity on T2-weighted MRI both in vitro and in vivo. EGFP fluorescent imaging could also be detected in the subcutaneous and intracranial glioma xenografts. Moreover, the expression of the transferritin receptor was significantly increased in glioma cells carrying the hFTH reporter gene.
Conclusion
Our study illustrated that hFTH generated cellular MR imaging contrast efficiently in glioma via regulating the expression of transferritin receptor. This might be a useful reporter gene in cell tracking and MR molecular imaging for glioma diagnosis, gene therapy and tumor metastasis.
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Acknowledgements
We thank Prof. XZ Chen for evaluating the MR molecular imaging (Radiology Department, Tiantian Hospital affiliated with Capital Medicine University). We also thank Prof. L Luo, GL Li, and L Xu (Pathological Department, Beijing Neurosurgical Institute) for the histological analysis of glioma specimens.
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This work was supported by a Grant from the National Natural Science Foundation of China (Nos. 81271563, 81372354, 81302186 and 81672478), the Beijing Natural Science Foundation (No. 7151002), the Beijing Health System High-level Personnel Building Foundation (No. 2013-3-018), the Beijing Laboratory of Biomedical Materials Foundation (PXM2014_014226_000005), and the Beijing Municipal Administration of Hospitals’ Youth Programme (QML20150505).
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The authors declare that they have no conflict of interest.
Ethical approval
Animal research was carried out in accordance with the recommendations in the Guide for the Care and Use of Laboratory Animals of the Beijing Tiantan Hospital, Capital Medical University. The experimental protocol was approved by the Committee on the Ethics of Animal Experiments of the Beijing Tiantan Hospital, Capital Medical University. All efforts were made to minimize the suffering of animal subjects.
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S. Cheng and R. Mi are co-authors.
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432_2017_2356_MOESM1_ESM.tif
S1 Fig. Western blot and histological analysis of intracranial glioma xenografts. (A) Expression of hFTH and EGFP in intracranially implanted xenografts was confirmed by western blot. (B) Prussian blue staining shows more iron accumulation in hFTH glioma xenografts (400×). (C) Immunofluorescent analysis of implanted xenografts expressing hFTH (red) or EGFP (green) alone (400×) (TIF 7386 KB)
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Cheng, S., Mi, R., Xu, Y. et al. Ferritin heavy chain as a molecular imaging reporter gene in glioma xenografts. J Cancer Res Clin Oncol 143, 941–951 (2017). https://doi.org/10.1007/s00432-017-2356-z
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DOI: https://doi.org/10.1007/s00432-017-2356-z