Chinese Journal of Magnetic Resonance ›› 2004, Vol. 21 ›› Issue (3): 289-294.

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ANALYSIS OF EXPRESSION OF CYTOSINE DEAMINASE GENE IN COLON CANCER CELLS WITH 19F MAGNETIC RESONANCE SPECTROSCOPY

HUANG Jian-Hua1, WANG Yao1, LI Chong-Hui2, QIU Ben-Sheng3, ZHONG Da-Guang1, LV Hai-Yan1   

  1. 1.Department of Immunoloy, Institute of Basic Medical Sciences; 2.Institute of Hepatobiliary Surgical Research; 3.PLA Medical Imaging Center, PLA General Hospital, Beijing 100853, China
  • Received:2004-02-13 Revised:2004-03-22 Online:2004-09-05 Published:2004-09-05
  • Supported by:

    国家自然科学基金资助项目(30070236).

Abstract:

To investigate the feasibility of using 19F-NMR spectroscopy to monitor cytosine deaminase (CD) transgene expression in human colon cancer cell line. The SW1116 cell line was transfected with a gene coding for CD from-E.coli- by the retroviral mediated method. The assessment of the expression of CD gene in the colon cancer cell line was carried out by RT-PCR, MTT assay of drug sensitivity to 5-FC and 19F-NMR spectroscopy. The results indicate that the transgenic SWCD2 cell line was more sensitive to the prodrug 5-fluorcytosine (5-FC) (IC50=66 μmol/L) as compared to the parent cancer cells SW1116 (IC50=16 mmol/L). After incubation of the cultured CD+ cells in 774 μmol/L 5-FC for 24 h, 19F-NMR spectra of the intact cells showed two broad resonances corresponding to 5-FC (taken up into cell chemical shift relative to trifluoroacetate of δ-91.7) and 5-FC produced by the action of CD (chemical shif of δ-93.3), respectively. These results demonstrated the feasibility of using 19F-NMR spectroscopy to noninvasively monitor CD gene expression in human colon cancer cells. This capability provides a new approach for measuring gene expression that will be useful in clinical gene therapy trials.

Key words: NMR, colon cancer cells, cytosine deaminase gene, 5-fluroscytosine, gene therapy

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