EB病毒潜伏膜蛋白LMP1编码基因沉默对NF—kB表达的影响

目的：探讨EBV阳性胃上皮细胞中潜伏膜蛋白1（latentmembraneprotein-1，LMP1）基因沉默对NF-kB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Westem-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；Westernblotting及免疫酶染色法检测LMP1基因沉默对NF—kB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；Westem blotting及免疫酶染色法检测结果显示LMP1沉默可干扰NF—KB表达，导致靶细胞核内NF-kB水平下降，细胞浆内水平升高，而NF—kB总蛋白有下降趋势。结论：LMP1基因特异性沉默能够影响NF-kB表达，抑制其核转移。

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

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.