摘要
目的观察性别决定区Y框蛋白2(Sox2)对表皮生长因子受体(EGFR)的调控及Sox2对胶质瘤细胞成球率的影响。方法构建表皮生长因子同源受体ErbB2、ErbB3、ErbB4启动子报告质粒,并用双报告基因检测分析方法检测Sox2对ErbB2、ErbB3、ErbB4表达的调控;Sox2表达质粒转染胶质瘤U251细胞,采用悬浮成球的方法培养U251细胞并计数成球率,观察U251细胞的自我复制;蛋白免疫印迹法(Western blot)检测EGFR和ErbB2蛋白的表达情况。结果 Sox2可以增加悬浮培养U251细胞的成球率(P<0.05);上调ErbB2、ErbB3、ErbB4启动子介导的荧光素酶蛋白表达活性,且表现出剂量依赖性。此外,Sox2可以提高EGFR和ErbB2受体的表达。结论 Sox2可以上调EGFR受体并促进胶质瘤干细胞的自我复制。
Objective To observe the regulation of sry-related high-mobility-group box-containing 2 （Sox2） on epidermal growth factor receptor（EGFR）and on the sphere formation rate of glioma stem ceils. Methods Promoter reporter plasmids of epidermal growth factor homologous receptor ErbB2 ,ErbB3, ErbB4 were respectively constructed. Sox2 expression plasmid was co-transfected together with the reporter plasmid into U251 cells. Then the luciferase activity was analyzed by dual-luciferase reporter assay system to test the regulation of Sox2 on ErbB2, ErbB3, ErbB4 promoters. Sphere formation assay was used to observe self renew of the cancer stem cells after transfection of Sox2. The expression of EGFR and ErbB2 proteins in the spheres was examined by Western blot. Results Sox2 could dose-dependently increase the ErbB2,ErbB3,ErbB4 promoter drived luciferase activity. Sox2 promotes the sphere-forming rate of U251 cells and upregulates the expression of EGFR and ErbB2 in the spheres. Conclusion Sox2 could up- regulate the expression of EGFR and promote selfrenew of glioma stem cells.
出处
《重庆医学》
CAS
CSCD
北大核心
2013年第34期4105-4107,4111,共4页
Chongqing medicine
基金
国家自然科学基金资助项目(30960150
31260246)
973计划前期研究专项课题(2011CBS512115
2012CB722408)
宁夏医科大学校级项目(XM200915
XT200909)
关键词
性别决定区Y框蛋白2
胶质瘤干细胞
受体
表皮生长因子
DNA复制
sry-related high-mobility-group box-containing 2
glioma stem cells
receptor,epidermal growth factor
DNA repliction
