鼻咽癌中EBV LMP1激活NF—κB的实验研究

目的：探讨在鼻咽癌细胞系中ＥＢ病毒潜伏膜蛋白ＬＭＰ１能否激活重要的核转录因子ＮＦ－κＢ，及ＮＦ－κＢ的活化对鼻咽癌细胞猕 表型的影响。方法：我们选用两种鼻咽癌细胞系即有ＬＭＰ１表达的ＣＮＥ－ＬＭＰ１和无ＬＭＰ１表达的ＨＮＥ１，用包含ＮＦ－κＢ结合位点萤虫素酶报道基因共转染的方法及软琼脂集落形成观察两株细胞中的ＮＦ－κＢ的活性。结果：建立稳定表达包含ＮＦ－κＢ结合位点萤虫素酶报道基因的两种细胞株。

ACTIVATION OF NFBBY EBV LIMP1 IN NPC

Objective: To determine the relationship between activation of NF-B and malignancy of NPC. Methods: Two NPC cell lines in vitro were selected as experimental materialsLMP1 positive CNELMP1 cell line and LMP1 negative HNE1 cell line. Four NPC clones H2, H5, C3 and C4 were established expressing HIVNFBluciferase reporter plasmid stably. LMP1 sense and LMP1 antisense plasmid were transferred into H2 and H5 from HNE1, and C3 and C4 from CNELMP1. H2, H5, C3 and C4 were treated by TPA at first. Then PDTC, the blocker of NFB, was used to treat NPC clones induced by TPA. Based on these findings determination of HNE1, H2 and CNELMP1, C4 was carried out to detect the influence of TPA and PDTC upon NPC cell malignant phenotype. Results: The activation of P65, an important subunit of NFB, was higher in CNELMP1 than that in HNE1 with western blotting and reporter gene analysis. The same colony rate was confirmed between HNE1 and its two clones H2 and H5, CNELMP1 and clones C3 and C4, but the colony rate of blot CNELMP1 and clones C3 and C4 was higher than that of HNE and clones H2 and H5. After being treated by TPA the colony rate rose. Conclusion: LMP1 can upregulate the activation of NFB, and such induce is in a dose and timedependent pattern. PDTC can inhibit NPC colony formation in agar, no matter whether NPC cells were treated by TPA or not. This indicates a strong relationship between NFB activation and NPC malignant phenotype, and provides an experimental basis for the use of PDTC as a drug for NPC chemical therapy in the future.