The Korean Journal of Gastroenterology (대한소화기학회지)

The Korean Society of Gastroenterology (대한소화기학회)
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Down-regulatory Effect of Interleukin-10 Gene Transfection for CXC Chemokines in Colonic Epithelial Cell

대장상피세포에서 Interleukin-10 유전자 전달의 CXC 케모카인에 대한 억제 효과

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
이국래;김찬규;김병관;장동경;이동호;김주성;정현채;이연;박종상;송인성

  • Published : 20030600
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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 3$\beta$[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.

목적: 염증성 장질환은 유발이나 지속에 있어 싸이토카인이 중요한 역할을 하는 것이 알려지면서 싸이토카인을 이용한 치료가 활발히 연구되고 있다. 그러나 싸이토카인 단백질을 직접 사용하는 경우 생체 이용률이 낮으며 낮은 약물동력학성과 화학적 불안정성이 문제되고 있으며 이에 싸이토카인 유전자 전달을 이용한 치료가 최근 도입되고 있다. 이에 본 연구에서는 배양된 생쥐의 대장암 세포주에서 비바이러스 유전자 운반체를 이용하여 IL-10 유전자를 전달하였을 때 IL-10이 발현하는지를 확인하고, 동시에 이렇게 전달된 IL-10이 염증성 자극으로 유발된 케모카인을 억제하는지를 알아보고자 하였다. 대상 및 방법: 치료 유전자와 플라즈미드는 pCAGGS mouse IL-10을 이용하고 유전자 운반체는 polyethyleneimine (PEI)과 3$\beta$ [L-ornithinamide-carbamoyl]cholesterol (O-chol)을 이용하였고, 생쥐의 대장암 세포주인CT26 세포를 이용하여 IL-10의 유전자 전달 이후 배양액에서의 IL-10 농도를 측정하였다. 이후 CT-26 세포를 lipopolysaccaride (LPS)나 TNF-$\alpha$ 로 자극한 경우, 케모카인인 KC나 MIP-2의 변화를 측정하였다. IL-10 유전자 전달로 IL-10이 발현된 CT-26 세포를 LPS나 TNF-$\alpha$ 로 처리하여 배양액에서 KC와 MIP-2를 측정, 비교하였다. 결과: PEI를 운반체로 이용한 경우 배양액의 IL-10 농도는 의미 있게 증가하였으나 O-chol을 이용한 경우에는 의미 있게 증가하지 않았다. CT-26 세포를 LPS나 TNF-$\alpha$ 로 처리하여 배양액에서 측정한 KC와 MIP-2는 모두 의미 있게 증가하였다. 이렇게 증가한 KC와 MIP-2는 PEI를 이용한 경우 의미 있게 감소하였으나 O-chol을 사용한 경우엔 MIP-2는 감소하지 않았고 KC는 LPS로 처리한 경우엔 감소하지 않았다. 결론: 생쥐의 대장세포에서 유전자 운반체를 이용한 IL-10 유전자 전달은 효율적으로 발현되었으며 이는 염증성 자극으로 증가된 케모카인을 효과적으로 억제하였으나 유전자 운반체에 따른 차이가 확인되었는데 이는 유전자 운반체의 효율성의 차이로 판단된다.

Keywords

References

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