摘要
目的研究FRK是否通过调节表皮生长因子受体(EGFR)-Y1173磷酸化促进胶质瘤细胞的凋亡。方法应用PolyJet^(TM)将FRK质粒转染入脑胶质瘤U251细胞中,western blot(WB)检测质粒转染效果及EGFR蛋白水平和EGFR-Y1173磷酸化水平的变化,流式细胞术检测脑胶质瘤细胞凋亡的变化;转染EGFR(WT)、EGFR(Y1173F)质粒,或共转FRK和EGFR(WT)、EGFR(Y1173F)质粒,流式细胞术检测脑胶质瘤U251细胞凋亡的变化。结果成功将FRK质粒转染入脑胶质瘤U251细胞,过表达FRK促进脑胶质瘤U251细胞的凋亡。过表达FRK增加EGFR-Y1173磷酸化水平,而对EGFR蛋白水平无影响。过表达EGFR(WT)、EGFR(Y1173F)质粒,细胞凋亡均减少,分别减少了25%和75%。WB结果显示,共转FRK和EGFR(WT)、EGFR(Y1173F)质粒成功;流式细胞术结果显示,与对照组相比,过表达FRK促进了细胞凋亡,EGFR抑制了细胞凋亡,且转染EGFR(Y1173F)质粒组的作用比转染EGFR(WT)的效果更明显。此外,转染EGFR(Y1173F)质粒可逆转过表达FRK对胶质瘤细胞的促凋亡作用。结论 FRK可以通过促进EGFR-Y1173磷酸化从而调节胶质瘤细胞的凋亡。
Objective To study whether FRK can promote the apoptosis of glioma cells by regulating phosphorylation of EGFR-Y1173. Methods The FRK plasmid was transfected into glioma U251 cells,transfection efficiency,the protein level and Y1173 phosphorylation of EGFR was exmined by western blot. The flow cytometry was used to examine the apoptosis of glioma cells. The EGFR（ WT） and EGFR（ Y1173F） plasmids were transfected to glioma U251 cells respectively,also the FRK,EGFR（ WT） and EGFR（ Y1173F） were co-transfected to glioma U251 cells,the apoptosis of glioma cells was tested by flow cytometry. Results The FRK plasmid was transfected into glioma U251 cells successfully,FRK over-expression promoted the apoptosis of glioma U251 cells. And FRK over-expression increased the EGFR-Y1173 phosphorylation,but had no effect on the protein level of EGFR. Also,the ability of glioma cells apoptosis was all decreased in EGFR（ WT） and EGFR（ Y1173F） over-expression group,and decreased by 25% and 75%,respectively. WB results showed,the FRK,EGFR（ WT） and EGFR（ Y1173F） plasmid were co-transfected into glioma U251 cells successfully. The flow cytometry results showed,the ability of glioma cells apoptosis was increased after FRK over-expression,but decreased in EGFR overexpression group,and the effect of EGFR（ Y1173F） on glioma cells apoptosis showed stronger than EGFR（ WT） ＇s. In addition,the promoting effect of FRK on glioma cells apoptosis was abolished after EGFR（ Y1173F） plasmid transfection. Conclusion FRK can regulate the glioma cells apoptosis by promoting phosphorylation of EGFR-Y1173.
作者
张道为
石琼
金戈
王军
宋旭
蔡畅
方震
周秀萍
郭克勤
于如同
ZHANG Dao-Wei SHI Qiong JIN Ge et al(The Graduate School, Xuzhou Medical College, Xuzhou 221002, China)
出处
《临床神经外科杂志》
CAS
2017年第1期21-25,共5页
Journal of Clinical Neurosurgery
基金
国家自然科学基金(81372699
81472345
81402046)
江苏省普通高校研究生科研创新计划项目(CXZZ13_0990)
