Down-regulatory Effect of Interleukin-10 Gene Transfection for CXC Chemokines in Colonic Epithelial Cell

Lee, Kook Lae; Kim, Chan Gyoo; Kim, Byeong Gwan; Chang, Dong Kyung; Lee, Dong Ho; Kim, Joo Sung; Jung, Hyun Chae; Lee, Yeon; Park, Jong Sang; Song, In Sung

Abstract

BACKGROUND/AIMS: Cytokine plays an important role in initiation and continuation of inflammatory bowel disease. However, cytokine protein has some limitation as a therapeutic tool because of low bioavailability, poor pharmacokinetics and chemical instability. Thus, we studied the effect of interleukin 10 (IL-10) gene transfection on murine colon cancer cell line by using non-viral gene carrier. METHODS: Therapeutic gene and plasmid was pCAGGS mouse IL-10 and gene carriers were polyethyleneimine (PEI) and 3beta[L-ornithinamide-carbamoyl] cholesterol (O-chol). After IL-10 gene transfection, we measured the level of IL-10 in supernatant of cultured CT-26 cells. The chemokine cytokine-induced neutrophil chemoattractant (KC) and macrophage inflammatory protein (MIP)-2, which were treated with lipopolysaccharide (LPS) or tumor necrosis factor-alpha (TNF-alpha), were measured after IL-10 gene transfection. RESULTS: The IL-10 values were increased significantly by using PEI, but not by using O-chol. The KC and MIP-2 values were increased when LPS or TNF-alpha were treated. When PEI was used, KC and MIP-2 values increased by LPS or TNF-alpha were decreased. When O-chol was used, the KC values increased by TNF-alpha were decreased but those treated by LPS were not decreased, and the MIP-2 values were not decreased. CONCLUSIONS: After IL-10 gene transfection in colon cancer cell, IL-10 cytokine was efficiently expressed. The increased chemokine values by LPS or TNF-alpha were suppressed by IL-10 gene transfection, but which was not constant because of carrier efficiency.