TSG101在人脑恶性胶质瘤中的表达与细胞增殖侵袭的关系

摘要

目的 探讨人肿瘤易感基因101(TSG101)在人脑恶性胶质瘤组织中的表达及其与胶质瘤细胞增殖侵袭的关系.方法 收集2015年7月至2016年10月我院神经外科收治的34例恶性胶质瘤患者的临床资料和肿瘤组织标本肿瘤组及同期因脑外伤入院的19例患者的正常脑组织作为对照组.使用Real-time PCR技术检测临床样本中TSG101的表达水平,Spearman等级相关分析检测TSG101的表达与患者各临床病理指标间的相关性.Western Bolt检测不同人脑恶性胶质瘤细胞系TSG101的表达水平,选取表达最高的细胞在体外构建干扰TSG101表达的细胞系siTSG101组,Western Bolt及Real-time PCR进行验证,MTT法检测细胞增殖能力,transwell侵袭实验检测细胞侵袭能力.结果 Western Bolt 结果示U251细胞中TSG101相对其他细胞高表达,体外成功干扰U251细胞TSG101的表达后其增殖能力在48 h、72 h出现升高(P＜0.05).Real-time PCR结果示恶性胶质瘤组织中TSG101的表达低于正常脑组织,差异具有统计学意义(P＜0.05);Spearman等级相关分析结果示TSG101表达水平与患者性别、年龄及体重无显著相关,与患者胶质瘤细胞分级存在负相关.transwell侵袭实验结果示干扰TSG101表达的细胞侵袭能力显著增强(P＜0.05).结论 TSG101在恶性胶质瘤组织中低表达,其表达与胶质瘤恶性程度呈负相关,并在胶质瘤的增殖侵袭过程中发挥抑制作用.%Objective To investigate the expression of human tumor susceptibility gene 101 (TSG101) in human malignant glioma and its relationship with the proliferation and metastasis of human glioma cells.Methods The clinical data and tumor tissues in 34 malignant glioma patients were collected from July 2015 to October 2016 in the department of neurosurgery in our hospital.The normal brain tissues in the 19 cerebral trauma patients were collected as control.PCR Real-time technology was used to detect the expression level of TSG101 in clinical samples.Spearman correlation analysis was used to detect the correlation between the expression level of TSG101 and the clinical pathological parameters of patients.Western Bolt was used to detect the expression level of TSG101 in the different human glioma cell line.The highest expression of TSG101 was selected to construct interference TSG101 expression cell lines,.Western Bolt and Realtime PCR was used to verify.The MTT method was used to detect the cell proliferation ability,and Transwell invasion assay was used to detect the ability of cell invasion.Results The results of Real-time PCR showed the expression of TSG101 in malignant glioma tissues was significantly lower than that in normal brain tissue (P＜0.05).Spearman correlation analysis showed that the correlation between expression level of TSG101 and the gender,age and body weight was not significantly,and the grade of patients with glioma cells was negatively correlated.The results of Western Bolt showed that TSG101 in U251 cells was highly expressed compared with that in other cells.After successfully interfered with the expression of TSG101 in U251cells in vitro,the proliferation ability of U251 siTSG101 cells was significantly increased in 48h and 72h (P＜0.05) after the expression of TSG101 in vitro.Transwell invasion assay showed significant enhancement of the cell invasion ability of interfering TSG101 expression (P＜0.05).Conclusion The expression of TSG101 in malignant glioma tissue is low.Its expression is negatively correlated with the degree of malignancy of gliomas.It plays an inhibitory role in the proliferation and invasion of glioma.