HSP70-inducible hNIS-IRES-eGFP reporter imaging: Response to heat shock Journal Article
| Authors: | Che, J.; Doubrovin, M.; Serganova, I.; Ageyeva, L.; Beresten, T.; Finn, R.; Blasberg, R. |
| Article Title: | HSP70-inducible hNIS-IRES-eGFP reporter imaging: Response to heat shock |
| Abstract: | A retroviral vector pQHSP70/hNIS-IRES-eGFP (pQHNIG70) was constructed containing the hNIS-IRES-eGFP dual-reporter genes under the control of an inducible human heat shock protein (HSPI70 promoter and RG2-pQHSP70/hNIS-IRES- eGFP (RG2-pQHNIG70) transduced cells were generated. Heat-induced expression of both reporter genes in RG2-pQHNIG70 cells was validated by enhanced green fluorescent protein (eGFP) fluorescence-activated cell sorter, in vitro radiotracer assays, and immunoblot and immunocytochemistry. A 2.2- to 6.1-fold (1311-), a 6.1- to 14.4-fold (99mTcO 4- ), and a 5.1- to 39-fold (fluorescence) increase above baseline was observed in response to graded hyperthermia (39-43°C). Increases in eGFP fluorescence and radiotracer uptake were first noted at 6 hours, reached a maximum at 24 hours, and fell toward baseline at 72 hours. A stable ratio of radiotracer uptake to eGFP fluorescence and to heat shock protein (HSP)70 protein was demonstrated over a wide range of expression levels, induced by different levels of heating. We also demonstrate that the local application of heat on RG2-pQHNIG70 xenografts can effectively induce hNIS and eGFP gene expression in vivo and that this expression can be efficiently visualized by fluorescence, scintigraphic, and micro-positron emission tomography imaging. Endogenous HSP70 protein and reporter expression was confirmed by postmortem tissue evaluations (immunoblot and immunohistochemistry) . The pQHIMIG70 reporter system can be used to study stress and drug responses in transduced cells and tissues. © 2007 BC Decker Inc. |
| Keywords: | genetics; positron emission tomography; methodology; proteins; mouse; animal; metabolism; animals; mice; gene expression; fluorescence; green fluorescent protein; pathology; tumor cells, cultured; diagnostic imaging; physiology; gene expression regulation; xenograft; cell culture; nude mouse; mice, nude; evaluation; reporter gene; transplantation, heterologous; green fluorescent proteins; binding site; binding sites; genes, reporter; radioactive tracers; heat shock protein 70; tracer; neoplasm transplantation; heat shock; hsp70 heat-shock proteins; ribosomes; cancer transplantation; enhanced green fluorescent protein; tissue culture; ribosome; regulatory sequence; regulatory elements, transcriptional; sodium iodide symporter; imaging systems; heat injury; heat; enhanced green fluorescent protein (egfp); cotransporter; heat stress disorders; symporters |
| Journal Title: | Molecular Imaging |
| Volume: | 6 |
| Issue: | 6 |
| ISSN: | 1535-3508 |
| Publisher: | Sage Publications, Inc. |
| Date Published: | 2007-11-01 |
| Start Page: | 404 |
| End Page: | 416 |
| Language: | English |
| DOI: | 10.2310/7290.2007.00036 |
| PUBMED: | 18053411 |
| PROVIDER: | scopus |
| DOI/URL: | |
| Notes: | --- - "Cited By (since 1996): 5" - "Export Date: 17 November 2011" - "Source: Scopus" |
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