Gene therapy of uterine leiomyoma: adenovirus-mediated herpes simplex virus thymidine kinase/ganciclovir treatment inhibits growth of human and rat leiomyoma cells in vitro and in a nude mouse model.
Uterine leiomyomas (LM) affect a high percentage of reproductive-age women. They develop as discrete, well-defined tumors that are easily accessible with imaging techniques--making this disease ideal for localized gene therapy approaches. In this study, we determined the efficacy of adenovirus-mediated herpes simplex virus thymidine kinase gene transfer in combination with ganciclovir (Ad-TK/GCV) as a potential therapy for LM. Rat ELT-3 LM cells and human LM cells were transfected with different multiplicity of infections (10-100 plaque forming units [PFU]/cell) of Ad-TK and treated with GCV (5, 10, or 20 microg/ml) for 5 days. To test the bystander effect, Ad-TK-transfected ELT-3 cells (100 PFU/cell) or LM cells (10 PFU/cell) were cocultured with corresponding nontransfected cells at increasing percentages and treated with GCV followed by cell counting. In ELT-3 cells transfected with Ad-TK/GCV (10, 20, 50, or 100 PFU/cell), the cell count was reduced by 24, 42, 77, and 87%, respectively, compared with the control cells (transfected with Ad-Lac Z/GCV). Similarly, in LM cells transfected with Ad-TK/GCV (10, 50, or 100 PFU/cell), the cell count was reduced by 31, 62, and 82%, respectively, compared with the control. A strong bystander effect was noted in both ELT-3 and LM cells with significant killing (p = 0.001) at a ratio of infected:uninfected cells of only 1:99 and maximal killing at 1:4. This study demonstrates the potential efficacy of the Ad-TK/GCV gene therapy approach as a viable nonsurgical alternative treatment for uterine LM.
Salama
S A
SA
Department of Obstetrics and Gynecology, University of Texas Medical Branch, Galveston, TX 77555-0587, USA.
Kamel
M
M
Christman
G
G
Wang
H Q
HQ
Fouad
H M
HM
Al-Hendy
A
A
D000256:Adenoviridae / Q000235:genetics*; D000818:Animals; D000998:Antiviral Agents / Q000627:therapeutic use; D024201:Bystander Effect; D045744:Cell Line, Tumor; D003131:Combined Modality Therapy; D018031:Connexin 43 / Q000032:analysis; D005260:Female; D015774:Ganciclovir / Q000627:therapeutic use; D015316:Genetic Therapy / Q000009:adverse effects / Q000379:methods*; D006801:Humans; D007889:Leiomyoma / Q000503:physiopathology / Q000628:therapy*; D051379:Mice; D008819:Mice, Nude; D051381:Rats; D011994:Recombinant Proteins / Q000627:therapeutic use; D018139:Simplexvirus / Q000235:genetics*; D013937:Thymidine Kinase / Q000009:adverse effects / Q000235:genetics / Q000627:therapeutic use*; D014162:Transfection; D014594:Uterine Neoplasms / Q000503:physiopathology / Q000628:therapy*
D000998:Antiviral Agents; D018031:Connexin 43; D011994:Recombinant Proteins; D013937:Thymidine Kinase; D015774:Ganciclovir