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背景与目的:LRIG1是由表皮生长因子(epidermalgrowthfactor,EGF)诱导产生、与细胞膜上的表皮生长因子受体(epidermalgrowthfactorreceptor,EGFR)特异性结合产生负反馈抑制效应的一种跨膜糖蛋白。本研究旨在通过研究LRIG1诱导神经胶质瘤细胞系H4的凋亡作用来探讨LRIG1对EGFR信号转导通路抑制效应的分子机制。方法:采用Lipofectamine介导的基因转染技术,将质粒p3XFLAG-CMV9-LRIG1转染神经胶质瘤细胞系H4,应用逆转录聚合酶链反应(reversetranscriptase-polymerasechainreaction,RT-PCR)和Westernblot方法检测转染后LRIG1和EGFR的mRNA与蛋白水平的变化,MTT法和流式细胞技术分析细胞的增殖和凋亡的变化。结果:转染p3XFLAG-CMV9-LRIG1组中LRIG1mRNA的表达水平(1.997±0.114)较未转染组(0.500±0.031)和转染空载体组(0.357±0.035)明显升高,差异有显著性(P<0.0001);LRIG1的蛋白表达(1.790±0.119)较未转染组(0.717±0.038)和转染空载体组(0.930±0.076)明显升高,差异有显著性(P<0.0001和P=0.001)。而EGFRmRNA的表达水平(0.463±0.033)较未转染组(1.157±0.067)和转染空载体组(0.933±0.058)明显降低,差异有显著性(P<0.0001和P=0.002);EGFR的蛋白表达(0.703±0.067)较未转染组(1.280±0.078)和转染空载体组(1.16
BACKGROUND & AIM: LRIG1 is a transmembrane glycoprotein that is induced by epidermal growth factor (EGF) and specifically binds to epidermal growth factor receptor (EGFR) on the cell membrane to produce negative feedback inhibition. This study aimed to investigate the molecular mechanism of LRIG1 on the EGFR signal transduction pathway by studying the role of LRIG1 in the apoptosis of glioma cell line H4. METHODS: The plasmid p3XFLAG-CMV9-LRIG1 was transfected into H4 cell line by Lipofectamine-mediated gene transfection technique. Reverse transcriptase-polymerase chain reaction (RT-PCR) The mRNA and protein levels of LRIG1 and EGFR were detected by RT-PCR. MTT assay and flow cytometry were used to analyze the changes of cell proliferation and apoptosis. Results: The expression level of LRIG1 mRNA in transfected p3XFLAG-CMV9-LRIG1 group was significantly higher than that in the untransfected group (0.500 ± 0.031) and the transfected empty vector group (0.357 ± 0.035) (1.997 ± 0.114) (P <0.0001). The protein expression of LRIG1 (1.790 ± 0.119) was significantly higher than that of the untransfected group (0.717 ± 0.038) and the transfected empty vector group (0.930 ± 0.076) 0.001). (0.463 ± 0.033) was significantly lower than that of untransfected group (1.157 ± 0.067) and transfected empty vector group (0.933 ± 0.058) (P <0.0001, P = 0.002) Compared with untransfected group (1.280 ± 0.078) and transfected empty vector group (1.16 ± 0.067)