LRIG1基因对胶质瘤SHG44的放疗增敏作用

LRIG1 gene enhances the radiosensitivity of glioblastoma SHG44 cells

目的研究亮氨酸重复序列免疫球蛋白样蛋白1(LRIG1)对人脑胶质瘤细胞系44细胞(SHG44)的放疗增敏作用。方法采用分子克隆技术,将构建的重组质粒p EGFP-C1-LRIG1转染SHG44,并稳定表达后在荧光显微镜下观察;逆转录聚合酶链反应(RT-PCR)技术研究放射照射后SHG44中目的基因LRIG1和表皮生长因子受体(EGFR)的信使核糖核酸(mRNA)表达情况;免疫组织化学法分析照射后SHG44中LRIG1和EGFR的蛋白表达差异;采用四甲基偶氮唑蓝(MTT)法分析干预后SHG44的细胞增殖能力;细胞侵袭实验(Transwell)分析干预后SHG44的肿瘤侵袭能力。结果重组质粒p EGFP-C1-LRIG1转染SHG44细胞可稳定表达LRIG1;SHG44+p EGFP-C1-LRIG1细胞相对于SHG44、SHG44+p EGFP-C1细胞,LRIG1与EGFR基因的mRNA和蛋白的表达水平有显著差异(P<0.01);且SHG44+p EGFP-C1-LRIG1细胞的细胞活性及侵袭能力较SHG44、SHG44+p EGFP-C1细胞明显下降(P<0.01)。结论在胶质瘤SHG44细胞中过表达LRIG1基因可通过下调EGFR表达,抑制SHG44细胞的恶性生物学行为,增加放疗敏感性。

Objective The effect of Leucine-rich repeats and immunoglobulin-like domain proteins（ LRIG1）on the radiosensitivity of glioblastoma cell line Suzhou Human Glioma Cells-44（ SHG44） was explored. Methods Molecular cloning technology was used to transfect the SHG44 cells with recombinant plasmid p EGFP-C1-LRIG1,and the stable expressed cells were observed under fluorescence microscope. Reverse transcription-polymerase chain reaction（ RT-PCR） technology was used to study the change of messenger RNA（ mRNA） level of LRIG1 and epidermal growth factor receptor（ EGFR） in the interventional glioma cell. Immunohistochemical method was used to investigate the different protein expressions of LRIG1 and EGFR. MTT method was used to analyze the cell activity of the interventional gliomas cells. Transwell invasion experiment was used to study the invasive changes of the interventional gliomas cells. Results SHG44 cells with recombinant plasmid p EGFP-C1-LRIG1 was transfected successfully with stable expression of LRIG1. There was significant difference between the SHG44 ＋ p EGFP-C1-LRIG1 and SHG44,SHG44 ＋ p EGFP-C1 in the expression of LRIG1 and EGFR（ P〈0. 01）. MTT method and Transwell study showed that the cell survival rate and invasion ability of the group of SHG44 ＋ p EGFP-C1-LRIG1 were much lower than those of group of SHG44 and SHG44 ＋ p EGFP-C1（ P〈0. 01）. Conclusion Overexpressed LRIG1 in the glioblastoma cell line SHG44 can inhibit the malignant behavior of SHG44 and enhance its radiosensitivity by down-regulating the expression of EGFR.