CAJ | 학술논문

目的探讨变异型IκBα(IκBαM)基因对人类多形性胶质母细胞瘤(GBM)细胞中尿激酶型纤溶酶原激活剂(urokinase type PA,uPA)和其受体(uPA-receptor,uPAR)表达的调控作用及其与肿瘤浸润行为的关系.方法构建质粒、基因转染以及IκBαM蛋白表达筛选,建立稳定表达IκBαM的人类GBM细胞株;用Transwell法测定肿瘤细胞的迁移能力;RT-PCR技术检测细胞内uPA、uPAR在RNA水平的表达;制作裸鼠皮下异位移植瘤生长模型,并采用免疫组化法分析肿瘤组织中uPA和uPAR的表达.结果 Transwell法测定G36△-M组肿瘤细胞的迁移能力明显降低;RT-PCR及肿瘤组化染色结果显示,uPA和uPAR在G36△-M组中的表达显著低于G36△、G36△-W和G36△-P三组,而在后三组间的表达无统计学意义.结论 IκBαM基因在RNA和蛋白水平均可显著降低人类恶性胶质瘤中uPA和uPAR的表达,进而减弱肿瘤细胞的浸润能力,抑制肿瘤组织的浸润.
목적 탐토변이형IκBα(IκBαM)기인대인류다형성효질모세포류(GBM)세포중뇨격매형섬용매원격활제(urokinase type PA,uPA)화기수체(uPA-receptor,uPAR)표체적조공작용급기여종류침윤행위적관계.방법 구건질립、기인전염이급IκBαM단백표체사선,건립은정표체IκBαM적인류GBM세포주;용Transwell법측정종류세포적천이능력;RT-PCR기술검측세포내uPA、uPAR재RNA수평적표체;제작라서피하이위이식류생장모형,병채용면역조화법분석종류조직중uPA화uPAR적표체.결과 Transwell법측정G36△-M조종류세포적천이능력명현강저;RT-PCR급종류조화염색결과현시,uPA화uPAR재G36△-M조중적표체현저저우G36△、G36△-W화G36△-P삼조,이재후삼조간적표체무통계학의의.결론 IκBαM기인재RNA화단백수평균가현저강저인류악성효질류중uPA화uPAR적표체,진이감약종류세포적침윤능력,억제종류조직적침윤.
Objective This study was designed to investigate the regulation of mutant-type IκBα (IκBαM) to Urokinase type plasminogen activator (uPA) and its receptor (uPAR) in human glioblastoma multiform(GBM) cells, and the correlation to tumor invasion. Method Human GBM cell line was used, transfected with IκBαM gene and selected IκBαM protein's expression, to establish cell line which steadily express IκBαM protein. The invasive activity of tumor cells was assayed in a transweli cell culture chamber, and the expression of uPA and uPAR in RNA level were analyzed by RT-PCR method. The cells were injected subcutaneously into the nude mice, and then compare the expression of uPA and uPAR by immunohistochemistry method. Result Transwell data shows that the invasive activity of G36△-M group cells was lower than the other three groups. The expression of uPA and uPAR in RNA and protein levels were decrease significantly in G36△-M group in vitro and in vivo, but the other three groups (G36△-W, G36△-P and G36△) almost same. Conclusions IκBαM gene could down-regulate the expressions of uPA and uPAR in the human GBM cell line in protein level, and reduce the ability of tumor invasion.