CAJ | 학술논문

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SHG-44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、160 μmol/L)的法舒地尔干预SHG-44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、48 h细胞增殖；伤痕愈合实验检测细胞迁移能力；Westernblot法检测ROCK-Ⅰ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)；MTr法显示法舒地尔对SHG-44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、160 μmol/L)与低浓度组(10、20 μmol/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.
목적 관찰Rho/ROCK신호전도통로억제제법서지이대인뇌효질류세포주SHG-44증식、천이급조망방면적영향병탐토기궤제.방법 사용불동농도(10、20、40、80、160 μmol/L)적법서지이간예SHG-44세포,작용불동시간,상차현미경하관찰세포형태변화；새서람(MTT)비색법검측16、24、48 h세포증식；상흔유합실험검측세포천이능력；Westernblot법검측ROCK-Ⅰ단백표체；류식세포의검측24h세포조망.결과 수착법서지이간예농도증가,ROCK-Ⅰ단백표체축점하강(P＜0.05)；MTr법현시법서지이대SHG-44세포증식적억제작용유명현적농도의뢰성급시간의뢰성(P＜0.05)；상흔유합실험현시교고농도조(40、80、160 μmol/L)여저농도조(10、20 μmol/L)、대조조비교,세포천이수도명현억제(P＜0.05)；류식세포의검측현시교고농도조조기조망솔고우대조조、저농도조(P＜0.05).결론 법서지이가능통과억제ROCK적활성급유도조망래억제인뇌효질류세포주SHG-44적증식、천이등악성생물학행위.
Objective To observe the anti-tumor effects of Rho/ROCK inhibitor,fasudil,on proliferation,migration and apoptosis of the human glioma cell line SHG-44,furthermore exploring the possible mechanism.Methods SHG-44 cells were treated with various concentrations (10,20,40,80 and 160 μmol/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 methyl thiazol tetrazolium (MTT) assay,and 50％ inhibition concentration was calculated.The migratory ability 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 160 μmol/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-44in vitro by inhibiting ROCK-Ⅰ and inducing apoptosis of tumor cells.