EB病毒潜伏膜蛋白LMP1编码基因沉默对NF-κB表达的影响  

The Influence of LMP1 Silence on Expression of NF-κ B in EBV Positive Gastric Cancer Cells

作　　者：李欣[1] 王笑峰[1] 刘霞[1] 冯艺[1] 罗兵[1] 

机构地区：[1]青岛大学医学院微生物教研室,山东青岛266021

出　　处：《现代生物医学进展》2009年第6期1040-1043,共4页Progress in Modern Biomedicine

基　　金：国家自然科学基金资助项目(30471623);山东省科技厅资助项目(2004GG3202016)

摘　　要：目的:探讨EBV阳性胃上皮细胞中潜伏膜蛋白1(latent membrane protein1,LMP1)基因沉默对NF-κB转录表达的影响。方法:以稳定表达LMP1的EBV阳性胃上皮细胞系作为靶细胞,采用化学合成的siRNA特异性沉默LMP1,用RT-PCR和West-ern-blotting分别检测其在不同时间段mRNA和蛋白水平特异性沉默效果;Western blotting及免疫酶染色法检测LMP1基因沉默对NF-κB转录表达及核转移的影响。结果:siRNA在mRNA和蛋白水平可特异性沉默LMP1的表达,且有时间效应关系;West-ern blotting及免疫酶染色法检测结果显示LMP1沉默可干扰NF-κB表达,导致靶细胞核内NF-κB水平下降,细胞浆内水平升高,而NF-κB总蛋白有下降趋势。结论:LMP1基因特异性沉默能够影响NF-κB表达,抑制其核转移。Objective： To study the effect of LMP 1 silence on expression of NF-kB in EBV positive gastric cancer cells. Methods： The chemically synthetic siRNA targeting LMP1 was shifted into EBV positive gastric carcinoma epithelial cell line by liposome 2000 at 50nM final concentration, The specific silence efficacy of LMP1 was estimated by RT-PCR and Western-blotting at different time. Immunoenzymatic staining and Western-blotting were used to demonstrate the expression of NF-kB influenced by LMP1 and nuclear transfer of NF-kB. Results： The results of RT-PCR and Western-blotting showed that specific siRNA could inhibit LMP1 expression at mRNA and protein level, besides the effect of siRNA on LMP1 time dependence. The results ofimmunoenzymatic staining and Western-blotting showed that the silence of LMP1 could interfere the expression of NF-kB, resulting in the expression of NF-kB down-regulated in cell nuclear and up-regulated in cytoplasm after siRNA targeting LMP1 transfection, and that the total protein of NF-kB in cell was downtrend. Conclusion： LMP1 specific silence could effect the expression of NF-kB and inhibit its nuclear transfer.

关 键 词：EB病毒 潜伏膜蛋白1 胃癌上皮细胞 NF-KB 小干涉RNA 

分 类 号：Q813[生物学—生物工程] R373[医药卫生—病原生物学]