Epstein-Barr virus-encoded latent membrane protein 1 modulates cyclin D1 by c-Jun/Jun B heterodimers

In our recent studies, we found that LMP1 encoded by Epstein-Barr virus could accelerate the formation of active c-Jun/Jun B heterodimer. We studied the regulation of cyclinD1 by c-Jun/Jun B heterodimers by laser scanning confocal influorescence microscopy, Western blot, luciferase activity assay, super-EMSA and flow cytometry in the Tet-on-LMP1 HNE2 cell line, in which LMP1 expression was regulated by Tet-on system. c-Jun/Jun B heterodimers induced by LMP1 could up regulate cyclin D1 promoter activity and expression. Overexpression of cyclinD1 accelerated the progression of cell cycle.     

EB病毒LMP-1在鼻咽癌细胞中通过JNK介导AP-1活化

 EB病毒潜伏膜蛋白 1 (latentmembraneprotein 1 ,LMP 1 )活化激活蛋白 1 (activatorprotein 1 ,AP 1 )信号传导途径与其致瘤作用密切相关 .为了探讨LMP 1活化AP 1信号传导的分子机制 ,在可诱导调控LMP 1表达的鼻咽癌细胞系L7中 ,首先通过荧光酶双报道系统确定了LMP 1表达能激活AP 1 ;在此基础上 ,用c JunPathDetect系统确定LMP 1表达活化AP 1是通过c Jun的磷酸化 (活化 )介导 .虽然LMP 1不能上调c Jun上游主要调节激酶c JunN端激酶 (c JunN terminalkinase ,JNK)的蛋白表达 ,但能显著促进JNK的磷酸化 (活化 ) ;在L7细胞中导入JNK相互作用蛋白 (JNK interactingprotein ,JIP)基因 ,抑制JNK的核移位能显著抑制LMP 1诱导的AP 1活化 ,同时对NFкВ活化也有部分抑制作用 .结果表明 ,EB病毒LMP 1在鼻咽癌细胞中通过JNK介导AP 1活化     

Nuclear translocation of EGF receptor regulated by Epstein-Barr virus encoded latent membrane protein 1

Epstein-Barrvirus(EBV),oneoftheDNAon-cogenicviruses,iscloselyassociatedwiththegenesisofBurkitt抯lymphoma,undifferentiatednasopharyn-gealcarcinoma(NPC),Hodgkin抯disease,gastriccancerand,lungcancer,etc.[1].EBVencodedlatentmembraneprotein1(LMP1)isconsideredtobethemajoroncogenicproteinofEBVencodedproteins.Biologicallyspeaking,LMP1isanintegralmembraneproteincontaining386aminoacids.Thethreedo-mains(CTAR1,CTAR2,CTAR3)intheC-terminusofLMP1havebeenshowntoinitiatethesignalingproc-ess.The…     

Nuclear accumulation of epidermal growth factor receptor and acceleration of G1/S stage by Epstein-Barr-encoded oncoprotein latent membrane protein 1

Epstein-Barr virus (EBV)-encoded latent membrane protein 1 (LMP1) is considered to be the major oncogenic protein of EBV-encoded proteins and has always been the core of the oncogenic mechanism of EBV. Advanced studies on nuclear translocation of the epidermal growth factor receptor (EGFR) family have greatly improved our knowledge of the biological function of cell surface receptors. In this study, we used the Tet-on LMP1 HNE2 cell line as a cell model, which is a dual-stable LMP1-integrated nasopharyngeal carcinoma (NPC) cell line and the expression of LMP1 which could be regulated by the Tet system. We found that LMP1 could regulate the nuclear accumulation of EGFR in a dose-dependent manner quantitatively and qualitatively. We also demonstrated that the nuclear localization sequence of EGFR played some roles in the location of the protein within the nucleus under LMP1 regulation and EGFR in the nucleus could bind to the promoters of cyclinD1 and cyclinE, respectively. We further demonstrated that EGFR is involved in the acceleration of the G1/S phase transition by LMP1 through binding to cyclinD1 and cyclinE directly. These findings provided a novel view that the acceleration of LMP1 on the G1/S transition via the nuclear accumulation of EGFR was critical in the process of nasopharyngeal carcinoma.     

Id1 interacts and stabilizes the Epstein-Barr virus latent membrane protein 1 (LMP1) in nasopharyngeal epithelial cells

The EBV-encoded latent membrane protein 1 (LMP1) functions as a constitutive active form of tumor necrosis factor receptor (TNFR) and activates multiple downstream signaling pathways similar to CD40 signaling in a ligand-independent manner. LMP1 expression in EBV-infected cells has been postulated to play an important role in pathogenesis of nasopharyngeal carcinoma. However, variable levels of LMP1 expression were detected in nasopharyngeal carcinoma. At present, the regulation of LMP1 levels in nasopharyngeal carcinoma is poorly understood. Here we show that LMP1 mRNAs are transcribed in an EBV-positive nasopharyngeal carcinoma (NPC) cell line (C666-1) and other EBV-negative nasopharyngeal carcinoma cells stably re-infected with EBV. The protein levels of LMP1 could readily be detected after incubation with proteasome inhibitor, MG132 suggesting that LMP1 protein is rapidly degraded via proteasome-mediated proteolysis. Interestingly, we observed that Id1 overexpression could stabilize LMP1 protein in EBV-infected cells. In contrary, Id1 knockdown significantly reduced LMP1 levels in cells. Co-immunoprecipitation studies revealed that Id1 interacts with LMP1 by binding to the CTAR1 domain of LMP1. N-terminal region of Id1 is required for the interaction with LMP1. Furthermore, binding of Id1 to LMP1 suppressed polyubiquitination of LMP1 and may be involved in stabilization of LMP1 in EBV-infected nasopharyngeal epithelial cells.     

Upregulated expression of kappa light chain by Epstein-Barr virus encoded latent membrane protein 1 in nasopharyngeal carcinoma cells via NF-kappaB and AP-1 pathways

B lymphocytes are generally considered to be the only source of immunoglobulins. However, increasing evidence revealed that some human epithelial cancer cell lines, including nasopharyngeal carcinoma (NPC) cell lines, expressed immunoglobulins. Moreover, we previously found that expression of kappa light chain in NPC cells could be upregulated by EBV-encoded latent membrane protein 1 (LMP1). Here, Western blot and flow cytometric analysis of intracellular kappa staining indicated that upregulation of the expression of kappa was inhibited by using LMP1-targeted DNAzyme and that Bay11-7082 and SP600125, inhibitors of JNK and NF-kappaB, respectively, inhibited LMP1-augmented kappa light chain expression in NPC cells. LMP1-positive NPC cells expressing the dominant-negative mutant of IkappaBalpha (DNMIkappaBalpha) or of c-Jun (TAM67) exhibited significantly decreasing kappa production compared with their parental cells. These results suggest that LMP1 elevated kappa light chain through activation of the NF-kappaB and AP-1 signaling pathways. The present study provided some hints of possible mechanisms by which human cancer cells of epithelial origin produced immunoglobulins.     

Cells in G_2/M phase increased in human nasopharyngeal carcinoma cell line by EBV-LMP1 through activation of NF-кB and AP-1

Although previous studies showed that the principal oncoprotein encoded by Epstein-Barr virus, latentmembrane protein 1 (LMP1) 5 could induce the nasopharyngeal carcinoma cells in G_2/M phase increased, littleis known about the target molecules and mechanisms. The present study demonstrated that LMP1 couldinduce the accumulation of p53 protein and upregulate its transactivity in a dose dependent manner, whichresulted in the decrease of the kinase activity of cdc2/cyclin B complex and inducing arrest at G2/M phasethrough the activation of NF-κB and AP-1 signaling pathways, and the effect of NF-κB was more obviousthan that of AP-1. This study provided some significant evidence for further elucidating the molecularmechanisms that LMP1 had effects on the surveillance mechanism of cell cycle and promoting the survivalof transformed cells and tumorigenesis.     

Cells in G2／M phase increased in human nasopharyngeal carcinoma cellline by EBV-LMP1 through activation of NF-kB and AP-1

Although previous studies showed that the principal oncoprotein encoded by Epstein-Barr virus, latent membrane protein 1 (LMP1), could induce the nasopharyngeal carcinoma cells in G2/M phase increased, littleis known about the target molecules and mechanisms. The present study demonstrated that LMP1 couldinduce the accumulation of p53 protein and upregulate its transactivity in a dose dependent manner, whichresulted in the decrease of the kinase activity of cdc2/cyclin B complex and inducing arrest at G2/M phasethrough the activation of NF-kB and AP-1 signaling pathways, and the effect of NF-kB was more obviousthan that of AP-1. This study provided some significant evidence for further elucidating the molecular mechanisms that LMP1 had effects on the surveillance mechanism of cell cycle and promoting the survivalof transformed cells and tumorigenesis.