富含亮氨酸重复序列的免疫球蛋白样基因-3对GL15细胞增殖和侵袭的影响及分子机制的研究  

LRIG3 gene regulates biological activity of GL15 cell line

作　　者：席桂发[1] 吴群[1] 王宝峰[2] 郭东生[2] 陈高[1] 张建民[1] 雷霆[2] 

机构地区：[1]浙江大学医学院附属第二医院神经外科,浙江杭州310009 [2]华中科技大学同济医学院附属同济医院神经外科,湖北武汉430030

出　　处：《浙江大学学报（医学版）》2008年第5期444-450,共7页Journal of Zhejiang University(Medical Sciences)

基　　金：国家自然科学基金资助项目(NO:30500521,NO:30271332)

摘　　要：目的:探讨富含亮氨酸重复序列的免疫球蛋白样基因-3(LRIG3)对胶质母细胞瘤细胞系GL15细胞增殖和侵袭的影响及其作用机制。方法:用LRIG3-EGFP质粒转染GL15细胞,激光共聚焦显微镜观察LRIG3在细胞中的定位,MTT法和Transwell分析检测EGF和AG1478对转染后的细胞增殖和侵袭的影响,逆转录酶聚合酶链反应(RT-PCR)和Western blot分别检测EGF作用于转染后的GL15细胞EGFR和LRIG3 mRNA和蛋白表达的变化。结果:LRIG3主要在胞浆中表达,转染后的细胞增殖和侵袭性明显下降。EGF可双向调节GL15细胞的增殖。用100ng/ml的EGF分别作用于各实验组细胞后,不同的时间点细胞EGFR、LRIG3 mRNA和蛋白表达不同,随时间延长,EGFR的表达下降,LRIG3表达增加。结论:过表达LRIG3可以抑制胶质瘤细胞生长并使其侵袭性下降,这种抑制作用与EGFR的活性密切相关。LRIG3可能是基因治疗胶质母细胞瘤重要的靶基因。Objective： To investigate the effects of leucine-rich repeats and immunoglobulin-like domains 3（LRIG3）on the biological activity of glioblastoma cell line GL15. Methods： Glioblastoma GL15 cells were cultured and transfected with LRIG3-EGFP plasmid. The location of LRIG3 in GL15 cells was observed with confocal microscopy. The proliferation and invasiveness of GL15 cells were detected with methyl thiazolyl tetrazolium （MTT） and Transwell methods respectively; the expression of epidermal growth factor receptor （EGFR） and LRIG3 mRNA and protein were detected with reverse transcriptase-polymerase chain reaction （RT-PCR） and Western blot respectively. Results： After transfection with the plasmid LRIG-EGFP,LRIG3 fusion protein was found in cytoplasm of GL15 cells and cell proliferative and invasiveness were reduced. The expression of EGFR and LRIG3 varied with the duration of EGF treatment （100 ng/ml）：the expression of EGFR decreased while the expression of LRIG3 increased as time prolonged. Conclusion： LRIG3 can inhibit the proliferation and invasiveness of glioblastoma cells and may be used as a target gene in gene therapy of glioblastoma.

关 键 词：胶质母细胞瘤/病理学 免疫球蛋白类 表皮生长因子 亮氨酸 转染 细胞系 

分 类 号：R739.41[医药卫生—肿瘤]