title: Cell death of human oral squamous cell carcinoma cell line induced by herpes simplex virus thymidine kinase gene and ganciclovir.
abstract: Suicide gene therapy combining herpes simplex virus thymidine kinase gene (HSVtk) and ganciclovir (GCV) is one strategy for the treatment of head and neck squamous cell carcinoma (HNSCC). The purpose of this study is to determine the mechanism of cell death that occurs in suicide gene therapy using HSVtk and GCV and to assess the safety of that therapy. The human oral squamous cell carcinoma cell line SAS was treated with adenovirus vector containing HSVtk gene (AdHSVtk) and GCV in vitro. Morphological changes including chromatin condensation, cell shrinkage, blebbing of cell membrane, and ballooning formations were observed. Changes in the localization of phospholipids in the cell membrane were also observed. The results of flow cytometry showed a maximum of about 65% of cells in the early phase of apoptosis. In addition, DNA fragmentation was investigated using the TUNEL method in vivo. Nude mice (BALB/c AJD(-nu-), aged 4 weeks) were implanted with SAS and treated with AdHSVtk and GCV. Tumor sections were then observed. The treatment group was confirmed to have DNA fragmentation-positive cells. These results suggest that suicide gene therapy using AdHSVtk and GCV led to apoptosis of the oral squamous cell carcinoma cell line.
authors.affiliation: Department of Oral and Maxillofacial Surgery, Nagoya University Graduate School of Medicine, 65 Tsuruma-cho, Showa-ku, Nagoya 466-8550, Japan.
authors.forename: Masaya
authors.initials: M
authors.lastname: Nishikawa
authors.forename: Yasushi
authors.initials: Y
authors.lastname: Hayashi
authors.forename: Noriyuki
authors.initials: N
authors.lastname: Yamamoto
authors.forename: Takafumi
authors.initials: T
authors.lastname: Fukui
authors.forename: Hirokazu
authors.initials: H
authors.lastname: Fukuhara
authors.forename: Kenji
authors.initials: K
authors.lastname: Mitsudo
authors.forename: Iwai
authors.initials: I
authors.lastname: Tohnai
authors.forename: Minoru
authors.initials: M
authors.lastname: Ueda
authors.forename: Masaaki
authors.initials: M
authors.lastname: Mizuno
authors.forename: Jun
authors.initials: J
authors.lastname: Yoshida
mesh_terms: D000256:Adenoviridae / Q000235:genetics; D000818:Animals; D017209:Apoptosis; D002294:Carcinoma, Squamous Cell / Q000473:pathology*; D045744:Cell Line, Tumor; D002462:Cell Membrane / Q000378:metabolism; D002469:Cell Separation; D002843:Chromatin / Q000378:metabolism; D053938:DNA Fragmentation; D005434:Flow Cytometry; D015774:Ganciclovir / Q000627:therapeutic use*; D006801:Humans; D020287:In Situ Nick-End Labeling; D008051:Lipid Bilayers / Q000378:metabolism; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008856:Microscopy, Fluorescence; D009062:Mouth Neoplasms / Q000473:pathology*; D018139:Simplexvirus / Q000201:enzymology*; D013937:Thymidine Kinase / Q000235:genetics*
chemical_list: D002843:Chromatin; D008051:Lipid Bilayers; D013937:Thymidine Kinase; D015774:Ganciclovir