CAJ | 학술논문

目的：探讨沉默烯醇化酶（ ENO1）对人胶质瘤U87细胞黏附和侵袭的影响及其机制。方法构建ENO1基因沉默人胶质瘤U87细胞系，将细胞分为对照组、空载组、沉默组1和沉默组2（沉默组2的ENO1表达量低于沉默组1）。采用MTS方法观察ENO1基因沉默后对人胶质瘤U87细胞体外黏附能力的影响，采用Transwell方法观察细胞侵袭能力，采用流式细胞技术观察细胞周期，采用Western blot和Real-time PCR法检测CD44、基质金属蛋白酶2（MMP-2）、MMP-9、CDK1、p27、p53和p-AKT，通过液相芯片技术检测TGF-β、IL-1和IL-6，通过报告基因技术考察NF-κB和Twist转录活性。结果与对照组比较，沉默组1和沉默组2的细胞黏附和侵袭能力均降低（ P均＜0．05），细胞周期阻滞在G0/G1期，CD44、MMP-2/9、CDK1、p-AKT蛋白和mRNA表达水平均下降（P均＜0．05），p27、p53蛋白和mRNA表达水平均增加（P均＜0．05），TGF-β、IL-1β、IL-6降低（P均＜0．05），NF-κB和Twist转录活性降低（P均＜0．05）。结论沉默人胶质瘤U87细胞中ENO1表达可显著抑制肿瘤细胞的体外侵袭和黏附能力，其抑制作用可能与调节细胞周期和侵袭相关基因表达有关。
목적：탐토침묵희순화매（ ENO1）대인효질류U87세포점부화침습적영향급기궤제。방법구건ENO1기인침묵인효질류U87세포계，장세포분위대조조、공재조、침묵조1화침묵조2（침묵조2적ENO1표체량저우침묵조1）。채용MTS방법관찰ENO1기인침묵후대인효질류U87세포체외점부능력적영향，채용Transwell방법관찰세포침습능력，채용류식세포기술관찰세포주기，채용Western blot화Real-time PCR법검측CD44、기질금속단백매2（MMP-2）、MMP-9、CDK1、p27、p53화p-AKT，통과액상심편기술검측TGF-β、IL-1화IL-6，통과보고기인기술고찰NF-κB화Twist전록활성。결과여대조조비교，침묵조1화침묵조2적세포점부화침습능력균강저（ P균＜0．05），세포주기조체재G0/G1기，CD44、MMP-2/9、CDK1、p-AKT단백화mRNA표체수평균하강（P균＜0．05），p27、p53단백화mRNA표체수평균증가（P균＜0．05），TGF-β、IL-1β、IL-6강저（P균＜0．05），NF-κB화Twist전록활성강저（P균＜0．05）。결론침묵인효질류U87세포중ENO1표체가현저억제종류세포적체외침습화점부능력，기억제작용가능여조절세포주기화침습상관기인표체유관。
Objective To investigate the effect and mechanism of enolase-1 ( ENO1) silencing on the adhesion and invasion of human glioma U87 cells in vitro.Methods ENO1 silencing human glioma U87 cells were established , and then they were divided into the control group , no-load group, silenced group 1 and silenced group 2.The cell invasion and adhesion were detected by Transwell and MTS assay , and cell cycle was analyzed by flow cytometry .Real-time PCR and Western blot were used to detect the expression of CD 44, matrix metalloproteinase 2 (MMP-2), MMP-9, CDK1, p27, p53 and p-AKT.The levels of TGF-β, IL-1, IL-6 and IL-8 were measured simultaneously by multiplexed Luminex assay , and the transcription activities of NF-κB and Twist were detected by reporter gene assay .Results Compared with the control group, the invasion rate and adhesion abilities were decreased in the silenced groups 1 and 2 (all P<0.05), the cell cycle was arrested in G0/G1 phase, the protein and mRNA expression of p27 and p53 increased significantly ( all P<0.05), while the expression of CD44, MMP-2/9, CDK1 and p-AKT was down-regulated significantly (all P<0.05), the levels of TGF-β, IL-1βand IL-6 were decreased (all P<0.05), and the transcription activities of NF-κB and Twist were decreased (all P<0.05).Conclusion ENO1 silencing could inhibit the invasion and adhesion of human U 87 cells by regulating the related gene expression of cell cycle and invasion .