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目的:观察Snail蛋白在人脑胶质瘤组织中的表达情况,探讨Snail表达对人脑胶质瘤U251细胞侵袭的影响。方法:收集首都医科大学附属北京天坛医院及北京安贞医院神经外科手术切除的脑胶质瘤组织65例,应用免疫组织化学S-P法检测人脑胶质瘤组织中Snail的表达。体外化学合成Snail序列特异性小干扰RNA(Snail-siRNA),应用脂质体介导转染U251细胞;RT-PCR、Western blotting检测转染后U251细胞中Snail mRNA和蛋白、E-cadherin蛋白表达水平变化,并采用Transwell小室检测转染后U251细胞侵袭能力的变化。结果:与正常脑组织相比,人脑胶质瘤组织中Snail蛋白阳性表达率明显增强(66.2%vs0,P<0.01),并且Ⅰ～Ⅱ级的胶质瘤组织阳性Snail阳性率明显低于Ⅲ～Ⅳ级(44.8%vs83.3%,P<0.01)。Snail-siRNA转染抑制U251细胞中Snail mRNA和蛋白的表达。Snail-siRNA转染组U251细胞中E-cadherin蛋白的表达明显高于Ctrl-siRNA组与未转染组(0.64±0.21vs0.15±0.16,0.21±0.19,P<0.01)。Snail-siRNA转染显著抑制U251细胞的侵袭(87.0±2.4vs140.0±4.9,136.0±5.3;P<0.05)。结论:人脑胶质瘤组织高表达Snail蛋白,siRNA干扰Snail蛋白的表达可抑制胶质瘤U251细胞的侵袭。
OBJECTIVE: To observe the expression of Snail protein in human glioma and to investigate the effect of Snail expression on the invasion of human glioma U251 cells. Methods: Sixty-five neurosurgical resected gliomas were collected from Beijing Tiantan Hospital and Beijing Anzhen Hospital affiliated to Capital Medical University. The expression of Snail in human glioma tissues was detected by immunohistochemical S-P method. Snail-siRNA was synthesized in vitro and transfected into U251 cells by lipofectamine. The expression of Snail mRNA and protein and E-cadherin protein in U251 cells were detected by RT-PCR and Western blotting The changes of invasiveness of U251 cells were detected by Transwell chamber. Results: Compared with normal brain tissue, the positive rate of Snail protein in human glioma tissues was significantly higher (66.2% vs0, P <0.01), and the positive rate of Snail in grade Ⅰ ~ Ⅱ glioma tissues was significantly lower than that in normal brain tissues Ⅲ ~ Ⅳ grade (44.8% vs83.3%, P <0.01). Snail-siRNA transfection inhibits Snail mRNA and protein expression in U251 cells. The expression of E-cadherin in U251 cells transfected with Snail-siRNA was significantly higher than that in Ctrl-siRNA and untransfected cells (0.64 ± 0.21 vs 0.15 ± 0.16, 0.21 ± 0.19, P <0.01). Snail-siRNA transfection significantly inhibited the invasion of U251 cells (87.0 ± 2.4 vs. 140.0 ± 4.9, 136.0 ± 5.3; P <0.05). CONCLUSIONS: Snail protein is highly expressed in human glioma tissues. SiRNA knockdown of Snail protein can inhibit the invasion of glioma U251 cells.