Miller W E, Earp H S, Raab-Traub N
Department of Microbiology and Immunology, University of North Carolina School of Medicine, Chapel Hill 27599-7295, USA.
J Virol. 1995 Jul;69(7):4390-8. doi: 10.1128/JVI.69.7.4390-4398.1995.
The Epstein-Barr virus (EBV)-encoded LMP1 protein is an important component of the process of transformation by EBV. LMP1 is essential for transformation of B lymphocytes, most likely because of its profound effects on cellular gene expression. Although LMP1 is expressed in the majority of nasopharyngeal carcinoma (NPC) tumors, the effect of LMP1 on cellular gene expression and its contribution to the development of malignancy in epithelial cells is largely unknown. In this study the effects of LMP1 on the expression and tyrosine kinase activity of the epidermal growth factor receptor (EGFR) were investigated in C33A human epithelial cells. Stable or transient expression of LMP1 in C33A cells increased expression of the EGFR at both the protein and mRNA levels. In contrast, expression of the EGFR was not induced by LMP1 in EBV-infected B lymphocytes. Stimulation of LMP1-expressing C33A cells with epidermal growth factor (EGF) caused rapid tyrosine phosphorylation of the EGFR (pp170) as well as several other proteins, including pp120, pp85, pp75, and pp55, indicating that the EGFR induced by LMP1 is functional. LMP1 also induced expression of the A20 gene in C33A epithelial cells. In C33A cells, LMP1 expression increased the proliferative response to EGF, as LMP1-expressing C33A cells continued to increase in number when plated in serum-free media supplemented with EGF, while the neo control cells exhibited very low levels of viability and did not proliferate. Immunoblot analysis of protein extracts from nude mouse-passaged NPC tumors also demonstrated that the EGFR is overexpressed in primary NPC tumors as well as those passaged in nude mice. This study suggests that the alteration in the growth patterns of C33A cells expressing LMP1 is a result of increased proliferative signals due to enhanced EGFR expression, as well as protection from cell death due to LMP1-induced A20 expression. The induction of EGFR and A20 by LMP1 may be an important component of EBV infection in epithelial cells and could contribute to the development of epithelial malignancies such as NPC.
爱泼斯坦-巴尔病毒(EBV)编码的潜伏膜蛋白1(LMP1)是EBV介导的细胞转化过程中的一个重要成分。LMP1对B淋巴细胞的转化至关重要,这很可能是因为它对细胞基因表达有深远影响。尽管LMP1在大多数鼻咽癌(NPC)肿瘤中表达,但LMP1对细胞基因表达的影响及其在上皮细胞恶性肿瘤发生中的作用在很大程度上尚不清楚。在本研究中,我们在C33A人上皮细胞中研究了LMP1对表皮生长因子受体(EGFR)表达和酪氨酸激酶活性的影响。LMP1在C33A细胞中的稳定或瞬时表达在蛋白质和mRNA水平上均增加了EGFR的表达。相反,在EBV感染的B淋巴细胞中,LMP1并未诱导EGFR的表达。用表皮生长因子(EGF)刺激表达LMP1的C33A细胞会导致EGFR(pp170)以及其他几种蛋白质(包括pp120、pp85、pp75和pp55)迅速发生酪氨酸磷酸化,这表明LMP1诱导的EGFR具有功能。LMP1还诱导了C33A上皮细胞中A20基因的表达。在C33A细胞中,LMP1的表达增强了对EGF的增殖反应,因为当接种在补充有EGF的无血清培养基中时,表达LMP1的C33A细胞数量持续增加,而新霉素对照细胞的活力极低且不增殖。对裸鼠传代的NPC肿瘤蛋白质提取物的免疫印迹分析也表明,EGFR在原发性NPC肿瘤以及裸鼠传代的肿瘤中均过度表达。本研究表明,表达LMP1的C33A细胞生长模式的改变是由于EGFR表达增强导致增殖信号增加以及LMP1诱导的A20表达对细胞死亡的保护作用。LMP1对EGFR和A20的诱导可能是EBV在上皮细胞中感染的一个重要组成部分,并且可能促成诸如NPC等上皮恶性肿瘤的发生。