Gou Xin-Min, Chen Yan, Chen Xiao-Yi, Arrand John R
Department of Pathology, Guangdong Medical College, Zhanjiang, Guangdong, PR China.
Ai Zheng. 2003 May;22(5):481-5.
BACKGROUND & OBJECTIVE: It has been proved that Epstein- Barr virus (EBV) latent membrane protein 1 (EBV-LMP1) can induce the expression of matrix metalloproteinase-9 (MMP-9). This study was designed to investigate the effect of EBV-LMP1 on related factors of metastasis of nasopharyngeal carcinoma cell line CNE1.
Expression of MMP-9 was studied in human NPC cell lines cultured in vitro: CNE1 (well differentiated cell line of NPC) and CNE1-GL (CNE1 cell line transfected with an eukaryotic LMP1-expression plasmid) by SP immunohistochemistry and Western blot analysis. Cell-matrix adhesion assay was used to study the adhesive ability of CNE1-GL cells. The effects of LMP1 on the invasion and migration of CNE1 cells were investigated by transwell methods.
MMP-9 was expressed in both cell lines but the intensity of the staining was different. The positive rates of expression of MMP-9 in CNE1 and CNE1-GL cells were 30.2% and 98.2%, respectively (P< 0.05). The increased expression of MMP-9 was also shown in CNE1-GL cells by Western blot analysis. Cell-matrix adhesion assay showed that the adhesive ability of CNE1-GL with the matrix (mean A value: 1.2508+/-0.0711) was higher than that of CNE1 cell (mean A value: 0.9519+/-0.068) (P< 0.001). Invasion assay and migration assay showed that the invasion and migration of CNE1-GL cell were higher than those of CNE1 cells (P< 0.01).
The transfection of LMP1 can increase the expression of MMP-9 in CNE1 cells. Abilities of adhesion, migration, and invasion of CNE1 cell were induced by LMP1. It is suggested that MMP-9 may have a role in the LMP1-induced acceleration of invasion and metastasis of NPC cells.
已证实爱泼斯坦-巴尔病毒(EBV)潜伏膜蛋白1(EBV-LMP1)可诱导基质金属蛋白酶-9(MMP-9)表达。本研究旨在探讨EBV-LMP1对鼻咽癌细胞系CNE1转移相关因子的影响。
采用SP免疫组化和蛋白质印迹分析方法,研究体外培养的人鼻咽癌细胞系CNE1(鼻咽癌高分化细胞系)和CNE1-GL(转染真核LMP1表达质粒的CNE1细胞系)中MMP-9的表达。采用细胞-基质黏附试验研究CNE1-GL细胞的黏附能力。通过Transwell法研究LMP1对CNE1细胞侵袭和迁移的影响。
两种细胞系均表达MMP-9,但染色强度不同。CNE1和CNE1-GL细胞中MMP-9的阳性表达率分别为30.2%和98.2%(P<0.05)。蛋白质印迹分析也显示CNE1-GL细胞中MMP-9表达增加。细胞-基质黏附试验显示,CNE1-GL与基质的黏附能力(平均A值:1.2508±0.0711)高于CNE1细胞(平均A值:0.9519±0.068)(P<0.001)。侵袭试验和迁移试验显示,CNE1-GL细胞的侵袭和迁移能力高于CNE1细胞(P<0.01)。
LMP1转染可增加CNE1细胞中MMP-9的表达。LMP1可诱导CNE1细胞的黏附、迁移和侵袭能力。提示MMP-9可能在LMP1诱导的鼻咽癌细胞侵袭和转移加速中起作用。