论文部分内容阅读
[目的]探讨第二信使神经酰胺信号传导诱导鼻咽癌细胞 p21WAF1表达的机理。[方法]Westernblot法检测 p53、p21、Jun、NFκB基因表达 ,试剂盒检测SAPK/JNK的活性。[结果]在鼻咽癌CNE2细胞中阿霉素能刺激 p53表达 ,不能上调p21WAF1 的表达 ,可见在CNE2细胞中p53为功能异常型。C2 - 神经酰胺处理CNE2细胞后 ,可见p53蛋白呈现下降趋势 ;而 p21WAF1 蛋白在处理4、16小时时 ,p21WAF1 表达明显增高 ,但36小时后 p21WAF1 恢复到基础水平。在6.25μmmol/L、12.5μmmol/L、25μmmol/L的C2_神经酰胺处理24小时后p21WAF1 上升 ,而50μmmol/L的C2_神经酰胺处理24小时后 ,p21WAF1 开始下降。C2_神经酰胺处理CNE2细胞2小时后 ,JNK的活性开始增加 ,6小时时保持激活状态 ,12小时时下降。c_Jun/AP_1蛋白在神经酰胺处理后3小时开始明显增高 ,持续到12小时 ,24小时下降至基础水平 ;C2_神经酰胺对CNE2细胞中NFκB蛋白表达没有明显的变化。[结论]C2_神经酰胺在鼻咽癌细胞CNE2中 ,通过p53非依赖性途经诱导p21WAF1 的表达。神经酰胺激活JNK ,从而激活AP_1因子 ,可能是其诱导 p21WAF1 表达的机理
[Objective] To investigate the mechanism of second messenger ceramide signaling in inducing the expression of p21WAF1 in nasopharyngeal carcinoma cells. [Methods] The expressions of p53, p21, Jun and NFκB genes were detected by Western blotting and the activity of SAPK / JNK was detected by kit. [Result] Doxorubicin stimulated the expression of p53 in CNE2 cells of nasopharyngeal carcinoma, and failed to up-regulate the expression of p21WAF1. It can be seen that p53 is a dysfunctional type in CNE2 cells. After treatment of C2-ceramide on CNE2 cells, p53 protein showed a downward trend; p21WAF1 protein expression was significantly increased at 4 and 16 hours after treatment with p21WAF1 protein, but p21WAF1 returned to basal level after 36 hours. P21WAF1 increased 24 h after treatment with 6.25 μmmol / L, 12.5 μmmol / L and 25 μmmol / L of C2-ceramide, whereas p21WAF1 began to decrease 24 h after 50 μmmol / L of C2-ceramide treatment. After 2 hours of treatment with C2-ceramide, the activity of JNK began to increase, kept active at 6 hours, and decreased at 12 hours. c_Jun / AP_1 protein increased significantly 3 hours after ceramide treatment, lasting for 12 hours and decreasing to basal level 24 hours later; C2_-ceramide had no obvious change on NFκB protein expression in CNE2 cells. [Conclusion] C2_-Ceramide induced the expression of p21WAF1 in CNE2 cells through p53-independent pathway. Ceramide activates JNK, which in turn activates the AP_1 factor, probably by its mechanism of inducing p21WAF1 expression