Hai Jian, Lin Qi, Zhang Hao, Lu Yang, Yi Jing
Department of Neurosurgery, Tongji Hospital, Tongji University, Shanghai, China.
Neurol Res. 2009 Apr;31(3):274-9. doi: 10.1179/174313209X380919. Epub 2008 Nov 26.
Preliminary works have indicated that panaxydol possesses growth inhibition and induces differentiation in rat C6 glioma cells. However, the molecular mechanism underlying this differentiation remains unknown. We sought to investigate the role of cyclic adenosine monophosphate (cAMP) in cellular differentiation induced by panaxydol.
C6 cells were treated with panaxydol and various specific inhibitors, and the inhibition of cell growth was assessed by 3-(4,5-dimethyl-2-thiazolyl)-2,5-diphenyl tetrazolium bromide assay and homotransplantation in nude mice. Astrocytic processes were quantified under a phase-contrasted microscope. Glial fibrillary acidic protein expression and cell migration were carried out by Western blot and scratch-wound test, respectively. In addition, the intracellular cAMP concentration was measured by immunoassay.
Panaxydol induces the elevation of intracellular cAMP concentration in C6 cells. The effects of growth inhibition in vitro and in vivo and induction of differentiation in C6 cells by panaxydol could be inhibited by the cAMP inhibitor, Rp-adenosine 3',5'-cyclic monophosphothioate, but not by protein kinase A or protein kinase C specific inhibitors.
These results suggest that the cAMP-dependent pathway may regulate cellular proliferation, migration and differentiation in C6 glioma cells by panaxydol.
前期研究表明，人参二醇对大鼠C6胶质瘤细胞具有生长抑制作用并能诱导其分化。然而，这种分化背后的分子机制仍不清楚。我们试图研究环磷酸腺苷（cAMP）在人参二醇诱导的细胞分化中的作用。
用人参二醇和各种特异性抑制剂处理C6细胞，通过3-（4,5-二甲基-2-噻唑基）-2,5-二苯基溴化四氮唑法和裸鼠体内移植评估细胞生长抑制情况。在相差显微镜下对星形胶质细胞突起进行定量分析。分别通过蛋白质免疫印迹法和划痕试验检测胶质纤维酸性蛋白表达及细胞迁移情况。此外，通过免疫分析法测定细胞内cAMP浓度。
人参二醇可诱导C6细胞内cAMP浓度升高。cAMP抑制剂Rp-腺苷3',5'-环磷酸硫酯可抑制人参二醇在体外和体内对C6细胞的生长抑制作用及诱导分化作用，而蛋白激酶A或蛋白激酶C特异性抑制剂则无此作用。
这些结果表明，cAMP依赖性途径可能通过人参二醇调节C6胶质瘤细胞的增殖、迁移和分化。
