IL-2/HSVtk Gene Expression in Neuroblastoma Cell Line

Koo, Ja Min; Moon, Young Bae; Cho, Hyun Sang; Park, Chong Young

Abstract

PURPOSE: We investigated the efficacy of in vitro transduction of neuro 2a cells with interleukin-2 (IL-2) and herpes simplex virus-thymidine kinase (HSVtk) gene followed by assay of IL-2 and administration of the antiviral drug ganciclovir (GCV). METHPDS: The LNC/IL-2/IRES/HSVtk vectors were transferred in vitro into the murine neuro 2a-cell lines and the transduced cell lines were selected in G-418, 500 mug/mL, for 14 days. The neuro-2a cells transduced with 1x10(6) LNC/IL-2/IRES/HSVtk vector (neuro- 2a/IL-2 /HSVtk) were cultured in 6 well culture plates for 24 hours and we measured IL-2 concentrations of 1 x10(5) neuro-2a/IL-2/HSVtk by ELISA method. And also, 1x10(5) neuro- 2a/IL-2/HSVtk were cultured in 96 well culture plates in 10 mug/mL concentrations of GCV for 72 hours and then we measured the surviving cells 24 hours, 48 hours and 72 hours after GCV administration.
RESULTS
We selected 5 colonies of the neuro-2a/IL-2/HSVtk and expanded in G-418 contained media. The IL-2 concentration of the neuro-2a/ IL-2/HSVtk was from 78.0+/-8.7 IU/day to 92.1+/-7.5 IU/day. The cell numbers of neuro-2a/IL-2/HSVtk survived after GCV administration were 3.7+/-1.5x10(2), 4.4+/-0.4x10(3), 5.0+/-1.0x10(2), 1.0+/-0.2x10(4), and 1.5+/-0.5x10(2), respectively. And the cell numbers of LNCX transduced neuro-2a cells survived were 7.5+/-0.5x10(5).
CONCLUSION
In vitro transduction of neuro-2a cells with the LNC/IL-2/IRES/HSVtk vector expressed effectively IL-2 and HSVtk gene simultaneously.