Lack of bystander killing in herpes simplex virus thymidine kinase-transduced colon cell lines due to deficient connexin43 gap junction formation.
Hum Gene Ther
; 9(15): 2253-61, 1998 Oct 10.
Article
in En
| MEDLINE
| ID: mdl-9794209
ABSTRACT
The efficacy of herpes simplex virus thymidine kinase (HSV-TK) gene therapy for colorectal carcinoma has been investigated in an in vitro system. The magnitude and the mechanism of the HSV-TK bystander effect was determined in three human colon tumor cell lines HCT-116, HCT-8, and HT-29. Each HSV-TK(+) cell line was generated by stable transduction with a bicistronic retroviral vector containing the HSV-TK and neomycin resistance (neo) genes; each exhibited an IC50 for GCV of < or =4 microM. When GCV was added to HSV-TK(+) cells mixed with parental cells or known bystander-positive cell lines, no bystander killing was evident in the HT-29 or HCT-8 cells. Western blots detected the expression of the gap junction protein connexin43 (Cx43) in HCT-8 and HT-29 cells; however, immunolocalization studies indicated predominantly cytoplasmic staining of Cx43 and no cell surface staining in these cell lines. Stable transfection of HCT-8 and HT-29 cells with Cx43 resulted in increased levels of Cx43 expression with the same subcellular distribution as before, yet there was again no apparent bystander killing. In contrast, Cx43 expression was localized to the cell surface in the bystander-positive colon tumor cell line HCT-116. These results demonstrate that expression and proper surface localization of Cx43 gap junctions are necessary components of the bystander effect in human colon tumor cells. They also indicate that future combination gene therapy approaches using coexpression of HSV-TK and Cx43 genes may not be applicable to all tumor systems.
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Collection:
01-internacional
Database:
MEDLINE
Main subject:
Thymidine Kinase
/
Genetic Therapy
/
Simplexvirus
/
Colonic Neoplasms
/
Gap Junctions
/
Connexin 43
Limits:
Humans
Language:
En
Journal:
Hum Gene Ther
Journal subject:
GENETICA MEDICA
/
TERAPEUTICA
Year:
1998
Document type:
Article
Affiliation country:
Estados Unidos