法舒地尔阻断Rho／ROCK通路对胶质瘤细胞生长的影响

Rho/ROCK inhibitor, fasudil, suppresses glioma cell line SHG-44 progression in vitro

目的观察Rho／ROCK信号转导通路抑制剂法舒地尔对人脑胶质瘤细胞株SHG44增殖、迁移及凋亡方面的影响并探讨其机制。方法使用不同浓度（10、20、40、80、160μmol／L）的法舒地尔干预SHG4d细胞，作用不同时间，相差显微镜下观察细胞形态变化；噻唑蓝（MTT）比色法检测16、24、48h细胞增殖；伤痕愈合实验检测细胞迁移能力；Westernblot法检测ROCK—1蛋白表达；流式细胞仪检测24h细胞凋亡。结果随着法舒地尔干预浓度增加，ROCK-I蛋白表达逐渐下降（P〈0．05）；MTT法显示法舒地尔对SHG44细胞增殖的抑制作用有明显的浓度依赖性及时间依赖性（P〈0．05）；伤痕愈合实验显示较高浓度组（40、80、160μmol／L）与低浓度组（10、20μmol／L）、对照组比较，细胞迁移受到明显抑制（P〈0．05）；流式细胞仪检测显示较高浓度组早期凋亡率高于对照组、低浓度组（P〈0．05）。结论法舒地尔可能通过抑制ROCK的活性及诱导凋亡来抑制人脑胶质瘤细胞株SHG-44的增殖、迁移等恶性生物学行为。

Objective To observe the anti-tumor effects of Rho/ROCK inhibitor, fasudil, on pro- liferation, migration and apoptosis of the human glioma cell line SHG-44, furthermore exploring the possi- ble mechanism. Methods SHG-44 cells were treated with various concentrations （ 10, 20, 40, 80 and 160 mxmol/L） of fasudil for different durations respectively. Morphological changes of SHG-44 cells were observed under a convered phase microscope. The inhibitory rate of cell proliferation was detected by meth- yl thiazol tetrazolium （MTT） assay, and 50% inhibition concentration was calculated. The migratory abili- ty was evaluated by using wound healing assay. Apoptosis rate was assessed by flow cytometry （FCM）. The expression level of ROCK-I protein was detected by using Western blotting. Results Fasudil inhibited SHG-44 cell proliferation and migration, and induced cell apoptosis in a time- and concentration-dependent manner. There was significant difference between higher-concentration groups （40, 80, and 160μmol/L） and lower-concentration groups （ 10, and 20 μmol/L） or control group （ P 〈 0. 05 ）. The expression of ROCK-I was remarkably lower in higher-concentration group than in control group or lower-concentration group at the same time points （P 〈 0. 05 ）. Conclusion Fasudil may suppress the progression of SHG-44 in vitro by inhibiting ROCK-1 and inducing apoptosis of tumor ceils.