酪氨酸蛋白磷酸酶非受体6型蛋白对胶质瘤干细胞成瘤的影响  

The effect of PTPN6 on the tumorigenesis of glioma stem cells

作　　者：张松阳 黄浩浩 满江红[1] ZHANG Song-yang;HUANG Hao-hao;MAN Jiang-hong(National Center for Biomedical Analysis,Beijing 100850,China)

机构地区：[1]国家生物医学分析中心,北京100850

出　　处：《国际药学研究杂志》2020年第4期263-268,275,共7页Journal of International Pharmaceutical Research

基　　金：国家自然科学基金资助项目(81872408)。

摘　　要：目的通过研究酪氨酸蛋白磷酸酶非受体6型蛋白(PTPN6)在胶质瘤干细胞(GSC)成瘤方面发挥的作用,探索新的肿瘤治疗相关标志物或治疗靶点,从而为临床胶质瘤的靶向治疗提供依据。方法利用Sun Brain数据库的数据,对PTPN6在正常脑组织和多形性胶质母细胞瘤(GBM)组织中的表达水平进行分析,比较二者表达水平的差异,并利用TCGAGBMLGG数据库比较不同级别胶质瘤组织中PTPN6表达水平的差异;根据TCGA-GBM数据库以及Rembrandt数据库的数据进一步分析PTPN6表达水平的高低与脑胶质瘤患者预后之间的关联。通过低氧培养条件对GSC及其分化后的肿瘤细胞进行培养,对比分化前和分化后的GSC细胞中PTPN6表达水平的差异;在GSC中敲低PTPN6,观察敲低后对肿瘤球形成的影响;利用免疫荧光技术检测GSC中PTPN6的细胞定位。结果PTPN6基因在GBM组织中的表达水平(mRNA)要高于正常脑组织(P<0.01);PTPN6 mRNA表达水平随胶质瘤恶性程度的增高而升高(Ⅲ级与Ⅱ级比较,P<0.05;Ⅳ级与Ⅱ级比较,P<0.01;Ⅳ级与Ⅲ级比较,P<0.01);PTPN6 mRNA高表达患者的生存期显著低于低表达患者(P<0.01)。低氧培养的GSC中PTPN6表达水平明显提高,并且增加水平与低氧时间呈正相关;在GSC中敲低PTPN6显著抑制了肿瘤球的形成;PTPN6在GSC中定位于细胞核。结论PTPN6在GSC中的高表达是胶质瘤细胞生长的一个相关因素,提示高水平PTPN6有利于GSC成瘤。Objective To investigate the effect of tyrosine-protein phosphatase non-receptor type 6(PTPN6)on the tumorigenesis of glioma stem cells(GSC),and explore the new potential biomarkers or therapeutic targets of glioblastoma multiforme(GBM),so as to provide a reference for the targeted clinical treatment of GBM.Methods The Sun Brain database was used to analyze the difference in the expression of PTPN6 gene(mRNA level)between the normal brain and GBM tissues.The TCGA-GBMLGG database and Rembrandt glioma database were used to compare the expression of PTPN6 in the different grade gliomas and the correlation between PTPN6 expression with prognosis of glioma patients.The PTPN6 expression was examined and compared between GSC and their differentiated tumor cells after cultivation of the cells under hypoxic conditions.Tumor sphere formation was examined after the PTPN6 knocked-down in GSC.Immunofluorescence was used to assess the localization of PTPN6 in GSC.Results The expression of PTPN6 was significantly higher in GBM than in normal brain(P<0.01).The expression of PTPN6 was significantly increased with the degree of glioma malignancy(gradeⅢvs gradeⅡ,P<0.05;gradesⅡ-Ⅲvs gradeⅣ,P<0.01).Patients with high expression of PTPN6 showed significantly shorter survival time period than those with low PTPN6 expression(P<0.01).The PTPN6 expression in GSC was significantly increased in hypoxic conditions and the increased PTPN6 level was positively correlated with the time length for the exposure of the cells to the hypoxic conditions.The knockdown of PTPN6 in GSC significantly inhibited the formation of tumor sphere.PTPN6 was found to be localized in the nucleus of GSC.Conclusion PTPN6 is required for GSC growth and high expression of PTPN6 indicates the malignant progression in GBM.

关 键 词：酪氨酸蛋白磷酸酶非受体6型蛋白 胶质瘤干细胞 成瘤 

分 类 号：R730.231[医药卫生—肿瘤]