Transduction of a drug-sensitive toxic gene into human leukemia cell lines with a novel retroviral vector

A. Abe, T. Takeo, N. Emi, M. Tanimoto, R. Ueda, J. K. Yee, T. Friedmann, H. Saito

Research output: Contribution to journalArticle

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Abstract

To investigate the possibility of killing tumor cells by the expression of an exogenously introduced toxic gene, we have constructed a novel retroviral vector (LTRNL) which has the polyA signal deleted herpes simplex virus type 1 thymidine kinase (HSV1-tk) gene. The vector becomes toxic by treating cells expressing HSV1-tk with the antiherpetic drugs acyclovir or ganciclovir (GCV). Cells of the human leukemia lines (K562, MEG-01) were infected with this vector and two transduced cell lines (K562/LTRNL, MEG-01/LTRNL) were established. Southern blot analysis confirmed the integration of the HSV1-tk transgene in these cells and Northern blot analysis exhibited the expression of 4.8-kb viral mRNA containing the HSV1-tk gene. The MTT (3-(4,5- dimethylthiazol-2-yl)-2,5-diphenyl tetrazolium bromide) assay for the in vitro cytotoxic effects of GCV to these cells demonstrated that concentrations of about 2.5 μM for K562/LTRNL and 1.25 μM for MEG-01/LTRNL cells resulted in 50% inhibition of cell growth after 72 hr. Subcutaneous tumors of MEG-01/LTRNL in KSN nude mice, but not those of uninfected MEG-01 cells, showed durable regressions after exposure of the mice to 40 mg/kg of GCV given subcutaneously once a day for 15 days. This study indicates that the LTRNL-infected human leukemia cells exhibit inducible susceptibility to GCV.

Original languageEnglish
Pages (from-to)354-359
Number of pages6
JournalProceedings of the Society for Experimental Biology and Medicine
Volume203
Issue number3
Publication statusPublished - 1993
Externally publishedYes

Fingerprint

Ganciclovir
Poisons
Viruses
Leukemia
Genes
Cells
Cell Line
Thymidine Kinase
Human Herpesvirus 1
Pharmaceutical Preparations
Tumors
Acyclovir
Cell growth
Assays
Messenger RNA
thymidine kinase 1
Southern Blotting
Transgenes
Nude Mice
Northern Blotting

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

Transduction of a drug-sensitive toxic gene into human leukemia cell lines with a novel retroviral vector. / Abe, A.; Takeo, T.; Emi, N.; Tanimoto, M.; Ueda, R.; Yee, J. K.; Friedmann, T.; Saito, H.

In: Proceedings of the Society for Experimental Biology and Medicine, Vol. 203, No. 3, 1993, p. 354-359.

Research output: Contribution to journalArticle

Abe, A, Takeo, T, Emi, N, Tanimoto, M, Ueda, R, Yee, JK, Friedmann, T & Saito, H 1993, 'Transduction of a drug-sensitive toxic gene into human leukemia cell lines with a novel retroviral vector', Proceedings of the Society for Experimental Biology and Medicine, vol. 203, no. 3, pp. 354-359.
Abe, A. ; Takeo, T. ; Emi, N. ; Tanimoto, M. ; Ueda, R. ; Yee, J. K. ; Friedmann, T. ; Saito, H. / Transduction of a drug-sensitive toxic gene into human leukemia cell lines with a novel retroviral vector. In: Proceedings of the Society for Experimental Biology and Medicine. 1993 ; Vol. 203, No. 3. pp. 354-359.
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