Expression characteristics of tumor necrosis factor alpha inducible protein 3 in glioma and its effect on invasion and migration of glioma cell
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摘要:
目的 探讨肿瘤坏死因子α诱导蛋白3(TNFAIP3)在胶质瘤中的表达及其对胶质瘤细胞侵袭、迁移的影响。 方法 采用实时荧光定量法(qRT-PCR)检测人星形胶质细胞NHA和胶质瘤细胞株U87、U251、SNB19、T98和LN229中TNFAIP3的表达，以U87和SNB19为实验对象。使用阳离子脂质体Lipofectamine 2000介导小干扰RNA(siRNA)转染U87和SNB19以下调TNFAIP3的表达；采用Transwell检测下调TNFAIP3对U87及SNB19侵袭的影响；运用划痕实验检测下调TNFAIP3对U87及SNB19迁移的影响。 结果 和NHA(0.144±0.008)相比，TNFAIP3在U87(1.380±0.066)、U251(0.663±0.062)、SNB19(1.405±0.032)、T98(1.002±0.068)、LN229(0.667±0.027)中的表达均升高(均P＜0.05)。si-TNFAIP3转染U87及SNB19可下调TNFAIP3的表达，继续培养24 h，U87及SNB19侵袭和迁移能力均下降(均P < 0.05)。 结论 TNFAIP3在胶质瘤中高表达，TNFAIP3可促进U87及SNB19的侵袭和迁移，si-TNFAIP3可抑制胶质瘤细胞的侵袭和迁移。 -
关键词:
- 肿瘤坏死因子α诱导蛋白3 /
- 胶质瘤 /
- 侵袭 /
- 迁移
Abstract:Objective To investigate the expression of tumor necrosis factor alpha inducible protein 3 (TNFAIP3) in gliomas and its effect on the invasion and migration of glioma cells. Methods Real-time fluorescence quantitative polymerase chain reaction (qRT-PCR) was used to detect the expression of TNFAIP3 in normal human astrocytes NHA and glioma cell lines U87, U251, SNB19, T98 and LN229, respectively. U87 and SNB19 were used as experiments subjects. Lipofectamine 2000-mediated small interference RNA (siRNA) was used to transfect U87 and SNB19 to down-regulate the expression of TNFAIP3. The effect of down-regulated TNFAIP3 on the invasion of U87 and SNB19 was detected by Transwell, and the migration was detected by scratch test. Results Compared with NHA (0.144±0.008), the expression of TNFAIP3 in U87 (1.380±0.066), U251 (0.663±0.062), SNB19 (1.405±0.032), T98 (1.002±0.068) and LN229 (0.667±0.027) were increased (all P < 0.05). Si-TNFAIP3 transfection into U87 and SNB19 could down-regulate the expression of TNFAIP3 and continue to culture for 24 hours, the invasion and migration ability of U87 and SNB19 were decreased (all P < 0.05). Conclusion TNFAIP3 is highly expressed in glioma, which can promote the invasion and migration of U87 and SNB19, and si-TNFAIP3 can inhibit the invasion and migration of glioma cells. -
Key words:
- Tumor necrosis factor alpha inducible protein 3 /
- Glioma /
- Invasion /
- Migration
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图 1 不同级别胶质瘤中TNFAIP3的表达情况及总生存期比较
注：A为低级别胶质瘤和胶质母细胞瘤中TNFAIP3表达情况；B为低级别胶质瘤和胶质母细胞瘤总生存期；num(T)为肿瘤组，num(N)为正常脑组织组。与正常脑组织比较，aP＜0.05。
Figure 1. Comparison of TNFAIP3 expression and overall survival in different grade gliomas
图 2 TNFAIP3 mRNA在正常人星形胶质细胞NHA和5种胶质瘤细胞中的表达量比较
注：与正常人星形胶质细胞NHA比较，aP < 0.05。
Figure 2. Comparison of TNFAIP3 mRNA expression in normal human astrocyte NHA and five glioma cells
图 3 转染48 h后U87、SNB19细胞各组TNFAIP3 mRNA的表达量比较
注：A为转染后U87细胞各组TNFAIP3 mRNA的表达量比较；B为转染后SNB19细胞各组TNFAIP3 mRNA的表达量比较。与Blank组比较，aP < 0.05；与si-NC组比较，bP < 0.05。
Figure 3. Comparison of TNFAIP3 mRNA expression in U87 and SNB19 cells 48 h after transfection
图 4 下调TNFAIP3表达对U87、SNB19细胞侵袭能力的影响
注：A为下调TNFAIP3表达后U87细胞穿过Transwell小室的侵袭细胞数；B为下调TNFAIP3表达后SNB19细胞穿过Transwell小室的侵袭细胞数。与Blank组比较，aP < 0.05；与si-NC组比较，bP < 0.05。
Figure 4. Effect of down-regulation of TNFAIP3 expression on invasion ability of U87 and SNB19 cells
图 5 下调TNFAIP3表达对U87、SNB19细胞迁移能力的影响
注：A为下调TNFAIP3表达后U87细胞迁移水平的相对表达量；B为下调TNFAIP3表达后SNB19细胞迁移水平的相对表达量。与Blank组比较，aP < 0.05；与si-NC组比较，bP < 0.05。
Figure 5. Effect of down-regulation of TNFAIP3 expression on migration ability of U87 and SNB19 cells
表 1 各组U87和SNB19细胞穿膜细胞数比较(x±s, 个)
Table 1. The number of transmembrane cells of U87 and SNB19 cells in each group was compared(x±s, cases)
组别 n U87 SNB19 Blank组 6 395.333±12.257 409.000±11.045 si-NC组 6 366.667±11.470 378.667±10.656 si-TNFAIP3组 6 185.000±11.860ab 199.000±11.045ab F值 184.800 216.200 P值 ＜0.001 ＜0.001 注：与Blank组比较，aP＜0.05；与si-NC组比较，bP＜0.05。 
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表 2 各组U87和SNB19细胞迁移率比较(x±s，%)
Table 2. Comparison of the migration rates of U87 and SNB19 cells in each group(x±s, %)
组别 n U87 SNB19 Blank组 6 82.954±2.343 85.500±2.114 si-NC组 6 76.197±2.160 75.510±2.556 si-TNFAIP3组 6 22.126±1.347ab 53.273±1.300ab F值 536.000 128.500 P值 ＜0.001 ＜0.001 注：与Blank组比较，aP＜0.05；与si-NC组比较，bP＜0.05。
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