论文部分内容阅读
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 investigate the effect of latent membrane protein 1 (LMP1) gene silencing on the transcription of NF-κB in EBV positive gastric epithelial cells. Methods: EBV-positive gastric epithelial cell lines stably expressing LMP1 were used as target cells. LMP1 was silenced by chemically synthesized siRNA. RT-PCR and West-ern-blotting were used to detect the mRNA and protein levels of LMP1 at different time points Silencing effect; Western blotting and immunoenzyme staining were used to detect the effect of LMP1 gene silencing on NF-κB transcription and nuclear translocation. Results: The expression of LMP1 at mRNA and protein level was silenced by siRNA, and there was a time-dependent effect. The results of West-ern blotting and immunoenzymatic staining showed that LMP1 silencing could interfere with the expression of NF-κB, leading to the increase of NF-κB Level decreased, the level of cytoplasm increased, while NF-κB total protein decreased. Conclusion: LMP1 gene-specific silencing can affect the expression of NF-κB and inhibit its nuclear translocation.