Retroviral Vector-mediated Interleukin-2 Gene Transfer into Human Renal Cell Carcinoma Cell Line

Chung, Byung Ha; Lee, Dong Hyeon; Hong, Sung Joo

Abstract

PURPOSE: The therapeutic dose of cytokine for patients with advanced renal cell carcinoma is very high and leads to toxic side effects and a substantial cost to the patients. Interleukin-2(IL-2) could be released continuously and slowly in the host by genetic engineering of IL-2 genes and increase host immunity with decreasing the a verse effects of the drug. We investigated the IL-2 gene expression, amplification of viral titer, and transduction of IL-2 gene into human renal cell carcinoma cell line with retroviral vectors.
MATERIALS AND METHODS
For the production of retroviral vectors with the IL-2 gene, we used PA-317 as a packaging cell and Caki-2 as a renal cell carcinoma cell. Retroviral supernatants were added to culture flask containing Caki-2 cells and after 48 hours, replacement with a media containing G418(Gibco, Grand Island, NY) 800 microgram/m1 was done for selection of transfected colonies. The selected colonies were cultured and then measured the amount of IL-2 production per 1xl0(6) for 24 hours using an ELISA assay kit(BioSource International, USA) for IL-2.
RESULTS
Thirteen colonies were selected and the amount of IL-2 production was 143.1 +/-75.3pg/m1/10(6) cells/24hr(range: 51.5-370.7).
CONCLUSIONS
The success of transduction of the IL-2 gene into human renal cell carcinoma cell lines with a retroviral vector will give a possibility in gene therapy for advanced renal cell carcinoma and may have promising results