法舒地尔阻断rhorock信号转导通路对人脑胶质瘤细胞生长影响的研究-effects of fasudil blocking rho rock signal transduction pathway on the growth of human glioma cells.docxVIP

法舒地尔阻断 Rho/ROCK 信号转导通路对 人脑胶质瘤细胞生长影响的研究摘要目的：研究Rho/ROCK信号转导通路抑制剂法舒地尔对人脑胶质瘤细胞系SHG44增殖、迁 移及凋亡方面的作用并探讨可能的作用机制。方法：使用不同浓度(10, 20, 40, 80, 160μmol/L)的法舒地尔干预人脑胶质瘤细胞系SHG44 细胞，作用不同时间后，相差显微镜观察细胞形态变化；MTT法检测16,24,48h细胞增殖情 况；伤痕愈合实验检测细胞迁移能力，划痕0h和24h分别拍照，使用Image J图像处理软件 计算迁移面积；流式细胞仪检测24h细胞凋亡情况。统计学数据处理采用SPSS 13.0软件处 理，组间比较采用ANOVA方差分析。结果：1、MTT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(P0.05)； 且16，24，48 h IC50分别为107.12,79.07,66.88μM/L(P0.05)，提示法舒地尔对SHG44细胞 增殖的抑制作用也具有时间依赖性；2、伤痕愈合实验显示随着法舒地尔干预浓度增加， SHG44细胞迁移受到明显抑制，较高浓度组（40，80，160μmol/L），与低浓度组（10, 20 μmol/L），对照组比较差异均有统计学意义(P0.05)； 3、流式细胞仪检测显示较高浓度 法舒地尔组（40，80, 160μmol/L），早期凋亡率明显增高，与对照组、低浓度组(10, 20 μmol/L )比较差异均有统计学意义(P0.05)。结论：法舒地尔可能通过抑制ROCK的活性，诱导凋亡来抑制人脑胶质瘤细胞系SHG44的 增殖、迁移等恶性生物学行为。因此，Rho/ROCK信号通路可能成为未来治疗胶质瘤的有 前景的靶点之一。关键词：法舒地尔，Rho/ROCK信号转导通路，增殖，迁移，凋亡Rho/ROCK Inhibitor, Fasudil, Suppresses Glioma Cell Line SHG44Progression in vitroAbstractObjected ： To explore the anti-tumor effects (proliferation, migration and apoptosis) of Rho/ROCK inhibitor, fasudil, including the possible mechanisms involved in the suppression of the human glioma cell line SHG44 progression.Method：After SHG44 were treated with various concentrations of fasudil(10, 20, 40, 80, 160μmol/L), the effects of ROCK inhibitor，fasudil, on proliferation, migration,and apoptosis of cultured tumor cells SHG44 were examined. Effect of Fasudil on the morphological changes of SHG44 cells was observed by phase microscope. MTT was performed to examine the effect of various concentrations of fasudil on the proliferation of glioma cells after treated in 16，24，48h. The wound healing assay was used to assess the effects of fasudil on the migration capacity of SHG44 cells. The sites of the wound line were photographed immediately after scratching and 24h later. The migration areas was measured by Image J. Annexin V/PI staining assay was used for investigating the ability of the fasudil to induce SHG44 cells apoptosis after 24h. Statistic package of SPSS 13.0 was used for the data analysis and significant d