GCV phosphates are transferred between HeLa cells despite lack of bystander cytotoxicity.

摘要: The role of gap junctional intercellular communication (GJIC) in bystander killing with herpes simplex virus thymidine kinase (HSV-TK) and ganciclovir (GCV) was evaluated U251 cells expressing a dominant-negative connexin 43 cDNA (DN14), HeLa cells, reportedly devoid protein. These cell lines both exhibited 0% GJIC when assayed by Lucifer Yellow fluorescent dye microinjection. Bystander cytotoxicity still apparent 50:50 cocultures DN14 HSV-TK-expressing but not cells. However, the sensitivity to GCV decreased nearly 100-fold (IC90=109 microM) cocultured compared results 100% cultures (IC90=1.2 microM). A more sensitive flow cytometry technique measure over 24 h revealed that detectable levels (29 23%, respectively). Transfer phosphorylated occurred within 4 after drug addition, triphosphate (GCVTP) accumulated 213+/-84 pmol/10(6) h. In GCVTP were coculture (867+/-33 cells) (1773+/-188 cells). half-life approximately four times measured (12.3 3.1 h, data suggest lack is due low transfer rapid Thus, phosphate non-HSV-TK-expressing may mediate either or sparing HSV-TK-positive depending upon specific metabolism.