缺氧诱导C6大鼠脑胶质瘤细胞亲环素A蛋白表达及其意义

为了探讨缺氧诱导C6大鼠脑胶质瘤细胞亲环素A(Cyclophilin A,CypA)表达水平的改变及其意义。本实验分别于正常(对照)及缺氧(缺氧组)环境培养C6细胞,利用蛋白印迹和RT-PCR技术检测CypA蛋白和mRNA的表达情况;在此基础上,利用RNA干扰技术和MTT技术检测抑制缺氧环境所诱导的CypA蛋白表达对C6细胞增殖的影响。结果显示:缺氧环境诱导C6细胞CypA mRNA和CypA蛋白表达上调,缺氧环境培养1h开始上调,12h达到顶峰;用CypA-siRNA干扰C6细胞CypA表达48h后,C6细胞CypA蛋白水平明显被抑制,细胞增殖能力较对照组显著下降(P<0.01)。本研究表明,缺氧可诱导C6细胞CypA表达水平上调,抑制CypA表达则可降低C6细胞的增殖能力。由此提示,CypA可作为胶质瘤治疗的新靶点。

The variation and its significance of the Cyclophilin A expression in the C6 rat glioma cells by hypoxia

To investigate the changes of the expression of Cyclophilin A (CypA) in the cultured C6 rat glioma cells after conditioned hypoxia,we cultured C6 cells in normal (control group) and hypoxia (hypoxia group) conditions,and examined the expression of CypA protein and mRNA by Western blot and RT-PCR technique. RNAi technique in vitro was used to suppress the expression of CypA gene,and the cell viability of each group was assayed by MTT colorimetry. The results showed that the expressions of CypA protein and mRNA increased after hypoxia and the highest level at 12 h after hypoxia. After 48 h transfection,the expression level of CypA protein in C6 cells was significantly inhibited by CypA-siRNA,and the cell viability also decreased significantly (P<0.01). The present results suggest that hypoxia might induce the increase of CypA expression in C6 cells,and the cell viability decrease significantly after inhibiting the expression of CypA. It implys that CypA gene may be one of the novel targets to inhibit growth of glioma cells.