S- and G2-phase cell cycle arrests and apoptosis induced by ganciclovir in murine melanoma cells transduced with herpes simplex virus thymidine kinase

Sung Jen Wei, Yee Chao, Yi Mei Hung, Wen Chang Lin, Den Mei Yang, Yung Luen Shih, Lan Yang Ch'ang, Jacqueline Whang-Peng, Wen K. Yang

Research output: Contribution to journalArticle

71 Citations (Scopus)

Abstract

Mechanism of cell killing by transfer of Herpes simplex virus type-1 thymidine kinase (HSVtk) and subsequent ganciclovir (GCV) treatment was examined in B16F10 murine melanoma model. While parental B16F10 melanoma cells were resistant to GCV at 100 μM or higher, HSVtk-transduced B16F10 melanoma cell clones became susceptible to GCV with IC50 of 0.1 to 0.3 μM. By means of various parameters including characteristic morphological changes, in situ DNA end-labeling, DNA ladder pattern, flow cytometric detection of sub-G1 DNA content, and annexin V binding of inverted cell surface phosphatidylserine, apoptosis was shown to be associated with the cell killing of ganciclovir on HSVtk-transduced melanoma B16F10 cells. Kinetic analysis showed that the signs of apoptosis were observed not until 60 h of continued GCV treatment and preceded first by a rise in p53 protein level in 12 h and then by S-phase/G2-phase cell cycle arrest associated with corresponding increases in the level of cyclin B1 protein but no apparent change in protein level of Bax or Cdc2. These results suggest that apoptosis occurred as a result of ganciclovir-induced cell cycle arrests rather than direct chemical effect on HSVtk-transduced B16F10 melanoma cells.

Original languageEnglish
Pages (from-to)66-75
Number of pages10
JournalExperimental Cell Research
Volume241
Issue number1
DOIs
Publication statusPublished - May 25 1998
Externally publishedYes

Fingerprint

G2 Phase Cell Cycle Checkpoints
Ganciclovir
Thymidine Kinase
G2 Phase
Simplexvirus
S Phase
Melanoma
Human Herpesvirus 1
Apoptosis
DNA
CDC2 Protein Kinase
Cyclin B1
bcl-2-Associated X Protein
Annexin A5
Phosphatidylserines
Cell Cycle Checkpoints
Inhibitory Concentration 50
Proteins
Clone Cells

Keywords

  • Apoptosis
  • Bystander effect
  • Cell cycle
  • GCV
  • HSVtk
  • Melanoma cells

ASJC Scopus subject areas

  • Cell Biology

Cite this

S- and G2-phase cell cycle arrests and apoptosis induced by ganciclovir in murine melanoma cells transduced with herpes simplex virus thymidine kinase. / Wei, Sung Jen; Chao, Yee; Hung, Yi Mei; Lin, Wen Chang; Yang, Den Mei; Shih, Yung Luen; Ch'ang, Lan Yang; Whang-Peng, Jacqueline; Yang, Wen K.

In: Experimental Cell Research, Vol. 241, No. 1, 25.05.1998, p. 66-75.

Research output: Contribution to journalArticle

Wei, Sung Jen ; Chao, Yee ; Hung, Yi Mei ; Lin, Wen Chang ; Yang, Den Mei ; Shih, Yung Luen ; Ch'ang, Lan Yang ; Whang-Peng, Jacqueline ; Yang, Wen K. / S- and G2-phase cell cycle arrests and apoptosis induced by ganciclovir in murine melanoma cells transduced with herpes simplex virus thymidine kinase. In: Experimental Cell Research. 1998 ; Vol. 241, No. 1. pp. 66-75.
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