姜黄素通过下调VEGF的表达抑制髓母细胞瘤Daoy细胞增殖的作用研究

发布时间：2018-06-18 20:11

  本文选题：姜黄素 + 髓母细胞瘤　； 参考：《重庆医科大学学报》2017年09期

【摘要】：目的:血管的新生与髓母细胞瘤的生长、侵袭和转移密切相关,本实验旨在观察姜黄素对髓母细胞瘤Daoy细胞的生长及其血管生长因子的影响,并探讨可能的机制。方法:体外培养髓母细胞瘤Daoy细胞,然后经不同浓度(5、10、20、40μmol/L)的姜黄素作用不同时间(24、48、72 h)后,通过MTT法检测姜黄素对Daoy细胞生长增殖的影响;ELISA法检测上清液中血管内皮生长因子(vascular endothelial growth factor,VEGF)的生成水平,Western blot检测Daoy细胞中Wnt/β-catenin通路中重要分子β-catenin以及VEGF蛋白水平的表达。结果:MTT结果示空白对照组、DMSO组、5μmol/L组、10μmol/L组、20μmol/L组和40μmol/L组作用24 h后,细胞的抑制率分别为(4.2±0.3)%、(4.3±0.5)%、(6.1±0.3)%、(6.9±0.7)%、(30.5±1.7)%和(40.9±2.4)%,差异具有统计学意义(F_(浓度)=151.997,P=0.000);当20、40μmol/L的姜黄素作用48 h和72 h后,Daoy细胞的增殖抑制率分别为(41.3±2.0)%、(51.2±2.9)%、(48.4±1.6)%和(60.9±3.2)%,相较于作用24 h,差异也具有统计学意义(F_(时间)=281.531,P=0.000;F_(浓度×时间)=269.728,P=0.000)。ELISA结果显示:空白对照组、DMSO对照组、5μmol/L组、10μmol/L组、20μmol/L组、40μmol/L组细胞上清液中VEGF的含量分别为(237.5±2.5)、(248.4±3.0)、(235.6±7.1)、(230.8±3.8)、(198.2±9.9)、(150.5±3.6)pg/m L,差异具有统计学意义(F_(浓度)=30.921,P=0.000)。当20μmol/L和40μmol/L的姜黄素作用48 h和72 h后,Daoy细胞上清液中生成的VEGF含量分别为(170.3±2.7)、(124.6±3.1)、(140.1±13.9)和(111.5±11.6)pg/m L,与作用24 h比较,差异也具有统计学意义(F_(时间)=69.234,P=0.000;F_(浓度×时间)=185.173,P=0.000)。Western blot结果显示,空白对照组、DMSO组、5μmol/L组、10μmol/L组、20μmol/L组和40μmol/L组Daoy细胞内β-catenin蛋白水平表达相对吸光度(absorbance,A)值分别为(0.654±0.021)、(0.873±0.034)、(0.869±0.018)、(0.857±0.024)、(0.472±0.035)和(0.351±0.019)(F_(浓度×时间)=27.813,P=0.000);VEGF蛋白水平的相对A值为(0.781±0.027)、(0.801±0.025)、(0.792±0.034)、(0.750±0.024)、(0.324±0.015)和(0.201±0.012)(F_(浓度)=74.826,P=0.000)。结果还显示,当姜黄素浓度为20μmol/L时,随着药物作用时间的延长,Daoy细胞内β-catenin和VEGF蛋白表达的相对A值明显降低,差异具有统计学意义,呈时间依赖性(F=19.156,P=0.001;F=100.282,P=0.000)。结论:姜黄素可通过下调Wnt/β-catenin通路及下游靶基因VEGF的表达进而抑制Daoy细胞增殖,这或将为姜黄素治疗髓母细胞瘤提供新的理论依据
[Abstract]:Aim: angiogenesis is closely related to the growth, invasion and metastasis of medulloblastoma. The aim of this study was to observe the effect of curcumin on the growth of Daoy cells and the angiogenesis factor of medulloblastoma, and to explore the possible mechanism. Methods: Daoy cells of medulloblastoma were cultured in vitro, and then treated with curcumin at different concentrations for 24 minutes and 872 hours after treatment with curcumin (40 渭 mol / L). The effect of curcumin on the proliferation of Daoy cells was detected by MTT assay. The level of vascular endothelial growth factor (VEGF) production in supernatant was detected by Elisa. The expression of 尾 -catenin and VEGFprotein in Wnt- / 尾 -catenin pathway in Daoy cells was detected by Western blot. Results after 24 h exposure to 20 渭 mol / L and 40 渭 mol / L groups of DMSO, 10 渭 mol / L and 40 渭 mol / L groups, 10 渭 mol / L group and 40 渭 mol / L group were treated for 24 h. The inhibitory rates of the cells were 4.2 卤0.3 and 4.3 卤0.5, respectively. The inhibition rates of Daoy cells were 41.3 卤2.030 卤2.90% and 30.5 卤1.7% and 40.9 卤2.4%, respectively. The difference was statistically significant (concentration of 151.997P0.000). The inhibitory rates of proliferation of Daoy cells at 2040 渭 mol / L for 48 h and 72 h were 41.3 卤2.00.30 卤2.92 卤2.90 卤1.60 卤1.6 and 6. 9 卤3.2, respectively, and the difference was statistically significant compared with the effect of 24 hours. The inhibitory rates of Daoy cells at 2040 渭 mol / L for 48 h and 72 h were 41.3 卤2.030% and 51.2 卤2.9%, respectively. The difference was statistically significant compared with the effect of 24 hours. 涔，

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