目的:研究Axin基因对神经胶质瘤细胞C6生物学特性及相关蛋白的影响。方法:采用噻唑蓝(MTT)比色法测定细胞生长曲线,应用流式细胞仪(FCM)测定细胞周期的变化,并用平板克隆形成实验对细胞克隆形成能力进行了检测。应用末端TdT酶标记技术(TUNEL)观察细胞凋亡及比率。采用免疫细胞化学染色法检测Ki67、CyclinD1、p53的表达。结果:转染Axin基因使细胞周期在G1期阻滞;细胞增殖能力和细胞克隆形成能力均显著降低;细胞凋亡率增高;Ki67及CyclinD1低表达,p53高表达。结论:转染Axin基因后胶质瘤细胞出现了一定的增殖抑制现象,诱导细胞凋亡,Axin基因可能通过上调p53基因表达和下调CyclinD1表达而抑制胶质瘤细胞的生长。
Objective:To observe the effects of Axin gene expression on the biological characteristics of glioma cells. Methods: The proliferation ability of transfected ceils was examined by MTT assay, cell growth curve and colony formation assay. The change of ceil cycle was analyses by flow cytometry ( FCM ). The apoptosis was measured by TUNEL staining. Immunostalning was used to detect the expression of Ki67, CyclinD1 and p53 in C6 cells and transfected cells. Results:The progression of the ceil cycle was arrested in G1 phase after transfected with Axin plasmids; the cell viability, growing speed and colony formation ability were decreased significantly at the same time; the apoptotic rate increased, the expressions of Ki67 and CyclinD1 decreased, but the p53 level increased. Conclusion:Axin gene could inhibit C6 cell proliferation and arrest the transfected cells in G1 phase, and this biological effect might be depended on the increased expression of p53 and the decreased expression of CyclinD1. The molecular mechanism of Axin in ceil proliferation needs more research.