人脑胶质瘤组织中EGFRvⅢ蛋白表达及其对细胞增殖的影响

Expression of EGFRvⅢ Protein in Human Glioma and Its Effect on Cell Proliferation

目的探讨人脑胶质瘤组织中表皮因子受体突变体Ⅲ(EGFRvⅢ)蛋白的表达及对细胞增殖的影响。方法应用免疫组化法(IHC)和酶联免疫吸附法(ELISA),检测54例人脑胶质瘤患者胶质瘤组织、癌旁组织中EGFRvⅢ表达情况;通过向胶质瘤U87细胞株中转染EGFRvⅢ的siRNA和阴性对照(NC)的siRNA进行靶向抑制,测定EGFRvⅢ基因对胶质瘤U87细胞增殖的影响;通过建立高表达EGFRvⅢ的胶质瘤U87细胞株,测定EGFRvⅢ基因对胶质瘤U87细胞增殖影响。结果胶质瘤组织中EGFRvⅢ表达量、阳性细胞率、染色强度评分、IHC评分高于癌旁组织(P<0.05);EGFRvⅢ-siRNA组细胞在培养24~96 h的OD450nm值显著低于NC-siRNA组(P<0.05或P<0.01);U87-EGFRvⅢ细胞中的EGFRvⅢ的相对表达量为(0.73±0.19),显著高于U87-control细胞中EGFRvⅢ蛋白的相对表达量(0.09±0.01)(P<0.01);U87-EGFRvⅢ组肿瘤体积及肿瘤质量分别为(0.51±0.24)cm^3、(1.12±0.15)g,均显著大于U87-control组的(0.09±0.03)cm^3、(0.39±0.20)g(P<0.01)。结论 EGFRvⅢ在胶质瘤组织中表达量较高,且能诱导胶质瘤U87细胞的增殖,高表达的EGFRvⅢ可促进胶质瘤U87细胞的成瘤能力,通过靶向抑制EGFRvⅢ后可降低胶质瘤U87细胞的增殖能力,EGFRvⅢ与胶质瘤的发生、发展密切相关,为靶向治疗胶质瘤提供科学依据。

Objective To investigate the expression of epidermal growth factor receptor variant type Ⅲ（ EGFRvⅢ） protein in human glioma and its effect on cell proliferation. Methods The expression of EGFRvⅢ in glioma tumor tissues and adjacent tissues of 54 cases of glioma patients were detected by immunohistochemical method（ IHC） and enzyme linked immunosorbent assay（ ELISA）; transfecting EGFRvⅢ siRNA and negative control（ NC） siRNA into U87 glioma cell line,and determine the effects of EGFRvⅢ gene on proliferation of U87 glioma cells; established the high expression of EGFRv Ⅲ in U87 glioma cell line,and determine the effects of EGFRvⅢ gene on proliferation of U87 glioma cells. Results The expression quanlity of EGFRvⅢ,percentage of positive cells,staining intensity score,IHC score in glioma tissues were higher than that of adjacent tissues（ P 〈0. 05）; when cultured 24 ~ 96 h the OD450 nmof EGFRvⅢ-siRNA group was significantly lower than that of NC-siRNA group（ P 〈0. 05 or P 〈 0. 01）; the relative expression of EGFRvⅢ in U87-EGFRvⅢ cells was（ 0. 73 ± 0. 19）,and was significantly higher than（ 0. 09 ± 0. 01） in U87-control cells（ P 〈 0. 01）; tumor volume and tumor weight in U87-EGFRvⅢ group were（ 0. 51 ± 0. 24）cm^3、（ 1. 12 ± 0. 15） g,and were significantly higher than that of（ 0. 09 ± 0. 03） cm^3、（ 0. 39 ± 0. 20） g in U87-control group（ P 〈0. 01）. Conclusion EGFRvⅢ is highly expressed in glioma tissues,which can induce U87 glioma cell proliferation,high expression of EGFRvⅢ can promote the tumorigenic ability of U87 glioma cells,EGFRvⅢ can inhibit the proliferation ability of U87 cells by target inhibiting the expression of EGFRvⅢ,EGFRvⅢ is closely related to the occurrence and development of glioma,which can provide scientific basis for targeted therapy of glioma.