Simultaneous imaging of temporal changes of NF-κB activity and viable tumor cells in Huh7/NF-κB-tk-luc2/rfp tumor-bearing mice

Wei Hsun Wang, I. Tsang Chiang, Yu Chang Liu, Fei Ting Hsu, Hong Wen Chen, Chuan Lin Chen, Yi Jang Lee, Wuu Jyh Lin, Jeng Jong Hwang

Research output: Contribution to journalArticle

3 Citations (Scopus)

Abstract

Few studies have reported that the effect of sorafenib on advanced human hepatocellular carcinoma (HCC) is taking place via the inhibition of NF-κB signal transduction. Here we constructed a human HCC Huh7 stable clone with NF-κB-responsive element to drive dual reporter genes, herpes simplex virus thymidine kinase (tk) and firefly luciferase (luc2), and co-transfected with a third red fluorescent protein (rfp) gene, renamed as Huh7/NF-κB- tkluc2/ rfp cells, and combined with bioluminescent imaging (BLI) and red fluorescent protein imaging (RFPI) to monitor the effect of sorafenib on NF-κB activation and tumor inhibition. The results show that sorafenib could suppress the NF-κB-DNA binding activity, and the expression of downstream effector proteins. Notably, the relative photon fluxes obtained from RFPI and BLI, which represent the viable tumor cells and cells with NF-κB activation, decreased after sorafenib treatment by 50 to 65%, and 87.5 to >90%, respectively, suggesting that NF-κB activation is suppressed in viable HCC cells by sorafenib. Simultaneous molecular imaging of the temporal change of NF-κB activity and of viable cells in the same Huh7/NF-κB-tk-luc2/rfp tumors of the animal may reflect the real status of NF-κB activity and the viable tumor cells at the time of imaging.

Original languageEnglish
Pages (from-to)339-350
Number of pages12
JournalIn Vivo
Volume27
Issue number3
Publication statusPublished - May 2013
Externally publishedYes

Fingerprint

Bearings (structural)
Thymidine Kinase
Tumors
Cells
Imaging techniques
Chemical activation
Hepatocellular Carcinoma
Neoplasms
Genes
B-Form DNA
Firefly Luciferases
Molecular imaging
Signal transduction
Molecular Imaging
Viruses
Simplexvirus
Reporter Genes
Photons
Animals
red fluorescent protein

Keywords

  • Human hepatocellular carcinoma
  • Imaging
  • RFP
  • Sorafenib
  • Temporal NF-κB monitoring

ASJC Scopus subject areas

  • Medicine(all)
  • Biochemistry, Genetics and Molecular Biology(all)
  • Pharmacology

Cite this

Wang, W. H., Chiang, I. T., Liu, Y. C., Hsu, F. T., Chen, H. W., Chen, C. L., ... Hwang, J. J. (2013). Simultaneous imaging of temporal changes of NF-κB activity and viable tumor cells in Huh7/NF-κB-tk-luc2/rfp tumor-bearing mice. In Vivo, 27(3), 339-350.

Simultaneous imaging of temporal changes of NF-κB activity and viable tumor cells in Huh7/NF-κB-tk-luc2/rfp tumor-bearing mice. / Wang, Wei Hsun; Chiang, I. Tsang; Liu, Yu Chang; Hsu, Fei Ting; Chen, Hong Wen; Chen, Chuan Lin; Lee, Yi Jang; Lin, Wuu Jyh; Hwang, Jeng Jong.

In: In Vivo, Vol. 27, No. 3, 05.2013, p. 339-350.

Research output: Contribution to journalArticle

Wang, WH, Chiang, IT, Liu, YC, Hsu, FT, Chen, HW, Chen, CL, Lee, YJ, Lin, WJ & Hwang, JJ 2013, 'Simultaneous imaging of temporal changes of NF-κB activity and viable tumor cells in Huh7/NF-κB-tk-luc2/rfp tumor-bearing mice', In Vivo, vol. 27, no. 3, pp. 339-350.
Wang, Wei Hsun ; Chiang, I. Tsang ; Liu, Yu Chang ; Hsu, Fei Ting ; Chen, Hong Wen ; Chen, Chuan Lin ; Lee, Yi Jang ; Lin, Wuu Jyh ; Hwang, Jeng Jong. / Simultaneous imaging of temporal changes of NF-κB activity and viable tumor cells in Huh7/NF-κB-tk-luc2/rfp tumor-bearing mice. In: In Vivo. 2013 ; Vol. 27, No. 3. pp. 339-350.
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abstract = "Few studies have reported that the effect of sorafenib on advanced human hepatocellular carcinoma (HCC) is taking place via the inhibition of NF-κB signal transduction. Here we constructed a human HCC Huh7 stable clone with NF-κB-responsive element to drive dual reporter genes, herpes simplex virus thymidine kinase (tk) and firefly luciferase (luc2), and co-transfected with a third red fluorescent protein (rfp) gene, renamed as Huh7/NF-κB- tkluc2/ rfp cells, and combined with bioluminescent imaging (BLI) and red fluorescent protein imaging (RFPI) to monitor the effect of sorafenib on NF-κB activation and tumor inhibition. The results show that sorafenib could suppress the NF-κB-DNA binding activity, and the expression of downstream effector proteins. Notably, the relative photon fluxes obtained from RFPI and BLI, which represent the viable tumor cells and cells with NF-κB activation, decreased after sorafenib treatment by 50 to 65{\%}, and 87.5 to >90{\%}, respectively, suggesting that NF-κB activation is suppressed in viable HCC cells by sorafenib. Simultaneous molecular imaging of the temporal change of NF-κB activity and of viable cells in the same Huh7/NF-κB-tk-luc2/rfp tumors of the animal may reflect the real status of NF-κB activity and the viable tumor cells at the time of imaging.",
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