RKI-1447可抑制卵巢癌CAOV-3细胞的迁移及侵袭能力

发布时间：2019-01-11 13:07

【摘要】：目的 :观察Ras同源性基因(Ras homology,Rho)-Rho相关卷曲螺旋形成蛋白激酶(Rho-associated coiledcoil-forming protein kinase,ROCK)信号通路抑制剂RKI-1447对卵巢癌CAOV-3细胞迁移及侵袭能力的影响,并探讨其可能的作用机制。方法:体外培养CAOV-3细胞,以不同浓度的RKI-1447(1、10、20、30、40和50μmol/L)处理CAOV-3细胞12 h后,采用CCK-8法检测RKI-1447对CAOV-3细胞增殖抑制率的影响,同时设未用任何药物处理的CAOV-3细胞为空白对照组。划痕实验以及Transwell小室侵袭实验检测RKI-1447(1和10μmol/L)对CAOV-3细胞迁移和侵袭能力的影响;分别采用实时荧光定量PCR法和蛋白质印迹法检测RKI-1447对CAOV-3细胞中Rho C m RNA及其蛋白表达的影响;同时采用蛋白质印迹法检测对RKI-1447肌球蛋白轻链2(myosin light chain 2,MLC-2)蛋白及其磷酸化蛋白表达的影响。结果:采用不同浓度的RKI-1447作用于CAOV-3细胞12 h后,细胞的增殖被明显抑制,抑制率呈剂量依赖性,半数抑制浓度(half inhibition concentration,IC50)约为20μmol/L。RKI-1447浓度为1和10μmol/L处理细胞12 h后,CAOV-3细胞的迁移及侵袭能力被显著抑制(P值均0.05);Rho C m RNA及蛋白的表达被明显下调(P值均0.05),磷酸化MLC-2蛋白的表达水平也被明显下调(P0.05)。结论 :RKI-1447对卵巢癌CAOV-3细胞的增殖、迁移及侵袭能力具有抑制作用,这可能与其调控Rho-ROCK信号通路中Rho C的表达以及MLC-2蛋白的磷酸化水平相关。
[Abstract]:Aim: to investigate the effects of RKI-1447, a signal pathway inhibitor of Ras homologous gene (Ras homology,Rho)-Rho associated crimp forming protein kinase (Rho-associated coiledcoil-forming protein kinase,ROCK), on the migration and invasion of ovarian cancer CAOV-3 cells. The possible mechanism of its action is also discussed. Methods: CAOV-3 cells were cultured in vitro. CAOV-3 cells were treated with different concentrations of RKI-1447 for 12 h. The effect of RKI-1447 on the proliferation inhibition rate of CAOV-3 cells was detected by CCK-8 assay. At the same time, the CAOV-3 cells which were not treated with any drugs were used as the blank control group. The effects of RKI-1447 (1 渭 mol/L and 10 渭 mol/L) on the migration and invasion of CAOV-3 cells were detected by scratch test and Transwell chamber invasion assay. The effect of RKI-1447 on the expression of Rho C m RNA and its protein in CAOV-3 cells was detected by real-time fluorescence quantitative PCR and Western blotting respectively. At the same time, the expression of RKI-1447 myosin light chain 2 (myosin light chain 2 MLC-2 protein and its phosphorylated protein were detected by Western blotting. Results: the proliferation of CAOV-3 cells was significantly inhibited after 12 h treatment with different concentrations of RKI-1447, and the inhibition rate was dose-dependent. The half inhibitory concentration of (half inhibition concentration, was found to be in a dose-dependent manner. The migration and invasion of CAOV-3 cells were significantly inhibited after 12 h treatment with IC50 (20 渭 mol/L.RKI-1447) and 10 渭 mol/L (P < 0. 05). The expression of Rho C m RNA and protein was significantly down-regulated (P < 0. 05), and the expression of phosphorylated MLC-2 protein was also down-regulated (P0.05). Conclusion: RKI-1447 can inhibit the proliferation, migration and invasion of ovarian cancer CAOV-3 cells, which may be related to the regulation of Rho C expression in Rho-ROCK signaling pathway and the phosphorylation of MLC-2 protein.
【作者单位】：贵州医科大学附属医院病理科;贵州省肿瘤医院病理科;
【基金】：国家自然科学基金资助项目(编号:81460482)~~
【分类号】：R737.31

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