나노방출제어시스템을 이용한 trichloroacetic acid와 epidermal growth factor 방출이 세포골격형성 유전자 발현에 미치는 영향 분석
Analysis of the effect of trichloroacetic acid and epidermal growth factor release on cytoskeleton gene expression using the nano-controlled releasing system
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목적: 본 연구에서는 나노방출제어시스템을 이용하여 trichloroacetic acid (TCA) 및 epidermal growth factor (EGF)를 인간치은섬유아세포에 적용하였을 때, 나타나는 액틴 세포골격과 관련된 유전자 발현의 변화 양상을 확인하고자 하였다. 재료 및 방법: TCA와 EGF가 조절방출될 수 있도록 만들어진 나노방출제어시스템을 이용하였다. 인간치은섬유아세포에 TCA만 적용된 군(EXP1), TCA와 EGF가 적용된 군(EXP2), 대조군(CON)의 3가지 군으로 나누어 48시간 배양하였다. Real-time PCR을 이용하여 액틴 세포골격과 관련된 유전자 26개의 발현 양상을 분석하였다. 피어슨상관관계분석을 통해 유전자들의 상관관계와 영향요인을 확인하였다. 결과: 액틴 세포골격과 관련된 유전자 26개 중 23개가 EXP1과 EXP2에서 상향조절되었고, 이 중 14개는 EXP1에 비하여 EXP2에서 유의미한 발현량 증가를 보였다. LPAR1은 EXP1에서만 하향조절되었고, GNA13은 EXP2에서만 상향조절되었고, F2R은 EXP2에서만 하향조절되었다. 액틴 단백질의 유전자 발현에 대하여 Rac1관련 유전자 중 3개와 CDC42가 가장 큰 영향요인으로 확인되었다. 결론: 인간치은섬유아세포의 액틴 세포골격 관련 유전자들은 나노방출제어시스템을 통하여 조절 방출된 TCA와 EGF에 의해 대부분 상향조절되었다.
Abstract
Purpose: Here, we verified that the actin cytoskeletal gene expression of human gingival fibroblasts was altered by the administration of trichloroacetic acid (TCA) and epidermal growth factor (EGF) using the nano-controlled releasing system. Materials and methods: The control and experimental groups were divided into 3 groups: the group with the TCA-only nano-controlled releasing system (EXP1), the group with the TCA- and EGF nano-controlled releasing system (EXP2), and the control group (CON) with 48-h incubation. Expression of 26 genes involved in the regulation of actin cytoskeleton were analyzed by real-time PCR followed by the determination of correlations and influential factors using the Pearson correlation analysis and multiple regression analysis. Results: Among 23 genes upregulated in EXP1 and EXP2, expression of 14 genes were significantly increased in EXP2 compared to EXP1. On the other hand, LPAR1 was downregulated only in EXP1, GNA13 was upregulated only in EXP2, and F2R was downregulated only in EXP2. Three Rac1-related genes and CDC42 were identified as the influential factors of the actin gene upregulation. Conclusion: The actin cytoskeleton genes in human gingival fibroblast were upregulated by the administration of TCA and EGF using HGC-based nano-controlled releasing system.
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