沙利度胺对胶质瘤U251细胞增殖及血管内皮生长因子表达的影响

Effects of thalidomide on proliferation and expression of vascular endothelial growth factor in glioma U251 cells

目的探究不同浓度沙利度胺对人胶质瘤U251细胞增殖和血管内皮生长因子(VEGF)表达的影响。方法用0,10,50,100,200μmol·L^(-1)沙利度胺分别处理人胶质瘤U251细胞,用噻唑蓝(MTT)法分检测24、48和72 h的细胞活力,用流式细胞术各组细胞凋亡情况,用蛋白质印迹(Western blot)法检测B淋巴细胞瘤-2(Bcl-2)蛋白表达的影响,用酶联免疫吸附(ELISA)法测定细胞培养上清液中VEGF水平。结果0,10,50,100,200μmol·L^(-1)沙利度胺组细胞凋亡率分别为(3.21±0.82)%,(28.21±4.23)%,(37.17±2.39)%,(68.12±3.28)%,(79.29±3.26)%,Bcl-2的表达分别为1.42±0.11,1.17±0.12,0.95±0.06,0.68±0.07,0.24±0.06,差异均有统计学意义(均P<0.05);干预72 h,0,10,50,100,200μmol·L^(-1)沙利度胺组细胞活性分别为0.89±0.15,0.56±0.08,0.43±0.26,0.22±0.19,0.14±0.12,VEGF水平分别为(395.63±9.47),(161.12±6.47),(112.52±6.16),(98.81±9.92),(79.92±3.48)pg·mL^(-1),差异均有统计学意义(均P<0.05),且具有剂量依赖性。结论沙利度胺对胶质瘤U251细胞具有显著的抑制作用,可以显著诱导细胞凋亡并抑制VEGF的表达。

Objective To investigate the effects of thalidomide on proliferation and vascular endothelial growth factor(VEGF) expression of human glioma U251 cells. Methods Human glioma U251 cells were treated with 0, 10, 50, 100, 200 μmol·L-1 thalidomide. MTT assay was used to detect the cell viability at 24, 48 and 72 h, flow cytometry was used to detect the cell apoptosis, the expression of B-cell lymphoma-2(Bcl-2) protein was detected by Western blot, the expression of VEGF in cell culture supernatant was detected by enzyme-linked immunosorbent assay(ELISA). Results The apoptosis rates in 0, 10, 50, 100, 200 μmol·L-1 thalidomide groups were(3.21±0.82)%,(28.21±4.23)%,(37.17±2.39)%,(68.12±3.28)%,(79.29±3.26)%,the expression of Bcl-2 were 1.42±0.11,1.17±0.12,0.95±0.06,0.68±0.07,0.24 ±0.06,all with significant difference(all P<0.05). After 72 h intervention, the cell viability in 0,10,50,100,200 μmol·L-1 thalidomide groups were 0.89±0.15,0.56±0.08,0.43±0.26,0.22±0.19,0.14±0.12,levels of VEGF were( 395. 63 ± 9. 47),( 161. 12 ± 6. 47),( 112. 52 ± 6. 16),( 98. 81 ± 9. 92)( 79. 92 ± 3. 48) pg·m L-1,all with significant difference( all P < 0. 05),and with dose-dependence. Conclusion Thalidomide has a significant inhibitory effect on glioma U251 cells,which can significantly induce apoptosis and inhibit the expression of VEGF.