摘要
背景与目的:LRRC4是作者最近克隆的一个新基因,该基因在原发性脑肿瘤活检标本中明显表达下调。本研究旨在研究LRRC4基因是否具有抑制脑肿瘤生长的能力。方法:LRRC4基因的全长编码区被亚克隆至表达载体pcDNA3.1中,应用脂质体转染的方法将重组的质粒载体导入胶质母细胞瘤细胞系U251,经G418筛选,建立稳定表达LRRC4基因的U251的细胞系。采用细胞增殖实验、软琼脂实验、肿瘤形成实验来考察LRRC4基因表达对于细胞生长和肿瘤形成的影响。结果:经过脂质体转染和筛选,建立了稳定表达LRRC4全长编码区的U251细胞系,用于进一步实验。比较未转染组和转染空白载体组,Northernblot实验证实转染了LRRC4基因的细胞LRRC4mRNA的表达增强。细胞增殖一定时间后,转染LRRC4基因的细胞较未转染细胞的生长速度明显减慢,克隆形成率明显降低。将这些细胞注射入无胸腺裸鼠体内,40天后处死裸鼠,测量肿瘤大小,结果显示转染LRRC4基因的细胞形成的肿瘤明显小于对照组。结论LRRC4基因可转染于人脑胶质母细胞瘤细胞系U251。LRRC4在U251细胞的表达有抑制瘤细胞增殖和抑制裸鼠移植瘤的形成和生长的作用。
BACKGROUND &OBJECTIVE:LRRC4is a novel g ene that the author has identified recently,whi ch displayed sig nificant downreg ul ation in primary brain tumor biopsies.This s tudy was desig ned to investig ate if LRRC4has the potential of suppressing bra in tumor g rowth.METHODS :The full-leng th coding reg ion of LRRC4g ene was subcl oned into the expression vector pcDNA3.1,the recombinant was intro duced into the g lioblastoma cell lin e U251by liposome transfection,and the U251cells stably expressing LRRC4g en e were established by G418selection.Furthermore,cell proliferation as say,soft ag ar assay,tumorig enesis assay wer e taken to examine the effect of LRRC4expression on cell g rowth and tumor f ormation.RESULTS :U251cells stably expressing full-leng th coding reg i on of LRRC4were established by lipofection-mediated transfectio n and selected for further study.Com pared with the nontransfected and vector-transfected cells ,the cells transfecte d with LRRC4cDNA exhibited a sig nificant increa se of expression of LRRC4mRNA by Northern blot analysis.Further,when cell pr oliferation was followed over sever al days ,the cells expressing the transfected LR RC4cDNA g rew more slowly than nontransfected cells.Consistently,the cells transfected with LRRC4e xhibited markedly lower colony formation rat e.These clones were injected into at hymic nude mice who was killed after 40days and the tumor sizes were evaluated.Tumor volume in mice was sig nificant ly smaller in the g roup of cells stabl y transfected with LRRC4cDNA than in t he control.CONCLUSION:LRRC4g ene may be transfected into the human g li oblastoma cell line U251.The expres sion of LRRC4in U251cells may have the poten tial to suppress tumor cell g rowth and the tumorig enesis of U251cell trans planted in nude mice.
出处
《癌症》
SCIE
CAS
CSCD
北大核心
2003年第9期897-902,共6页
Chinese Journal of Cancer
基金
国家自然科学基金项目(No.30100191,No.30270429)
国家-86-3计划(No.2001AA221031)
