过度表达LRIG1基因逆转人胶质瘤侵袭性的机制  被引量:26

Reverse effects of LRIG1 gene-overexpression on invasion of human glioma and its mechanism

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作  者:叶飞[1] 郭东升[1] 易伟[1] 牛洪泉[1] 舒凯[1] 万锋[1] 卢运萍[2] 雷霆[1] 

机构地区:[1]华中科技大学同济医学院附属同济医院神经外科,武汉430030 [2]华中科技大学同济医学院附属同济医院妇产科,武汉430030

出  处:《中华实验外科杂志》2005年第2期200-202,i003,共4页Chinese Journal of Experimental Surgery

基  金:国家自然科学基金资助项目 (30 2 71 332 )

摘  要:目的 探讨LRIG1基因抑制恶性胶质瘤细胞侵袭、浸润的分子机制。方法 应用脂质体介导的基因转染技术将p3XFLAG CMV9 LRIG1质粒转染入人神经胶质瘤H4细胞系 ,通过逆转录 聚合酶链反应 (RT PCR)和Westernblot方法检测转染前、后LRIG1和表皮生长因子受体(EGFR)基因mRNA和蛋白表达水平 ,用Boyden小室体外侵袭试验观察H4细胞通过人工重组基底膜能力的变化。结果 转染p3XFLAG CMV 9 LRIG1质粒后 ,H4神经胶质瘤细胞的LRIG1mRNA和蛋白表达均较对照组明显增强 (P <0 .0 1) ;而EGFRmRNA和蛋白表达均较对照组明显减弱 (P <0 .0 1)。LRIG1质粒转染的H4细胞穿过人工重组基底膜的相对百分率为 (13 .3± 1.8) % ,明显低于未处理组和转染空载体组的穿膜细胞百分率 (2 4.7± 1.8) % ,(2 4.0± 1.5 ) % (P =0 .0 0 8和P =0 .0 10 )。结论 EGFR的过表达促进了胶质瘤的侵袭、浸润 ;LRIG1通过抑制EGFR逆转了恶性胶质瘤侵袭、浸润。Objective To investigate the molecular mechanism of the inhibitory effect of LRIG1 on the invasion of malignant glioma.Methods Mediated by liposome,the plasmid p3XFLAG-CMV9-LRIG1 was transfected to neuroglioma cell line H4.Before and after transfection,the expression of mRNA and protein of LRIG1 and EGFR was detected by RT-PCR and Western blot.The invasive ability of H4 cell line in each group was compared by the method of Boyden chamber invasion assay.Results After H4 cell line was transfected with Plasmid p3XFLAG-CMV9-LRIG1,compared with control group,the expression of mRNA and protein of LRIG1 was obviously down-regulated (P<0.01) and that of EGFR was up-regulated (P<0.01).Boyden chamber in vitro invasion assay revealed that the mean invasion percentage of H4 cells that transfected with p3XFLAG-CMV9-LRIG1 plasmid ( 13.3± 1.8) % was significantly lower than that of untreated H4 cells and cells transfected with p3XFLAG-CMV9 plasmid ( 24.7± 1.8) %, (24.0± 1.5) % (P=0.008 and P=0.010).Conclusion The overexpression of EGFR accelerates invasion and infiltration of glioma.LRIG1 can reduce the invasive ability and infiltration of malignant glioma by inhibiting EGFR.

关 键 词:转染 逆转 恶性胶质瘤 EGFR 侵袭性 蛋白表达 对照组 人工重组 RNA 细胞 

分 类 号:R739.4[医药卫生—肿瘤]

 

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