[巨噬细胞移动抑制因子体外增强鼻咽癌细胞系的细胞迁移能力]

[Increased cell migration of nasopharyngeal carcinoma cell lines in vitro by macrophage migration inhibitory factor].

作者信息

Li Zhi, Lin Su-xia, Liang Ying-jie

机构信息

Department of Pathology, Sun Yat-sen Medical College, Sun Yat-sen University, Guangzhou 510089, China.

出版信息

Zhonghua Bing Li Xue Za Zhi. 2004 Feb;33(1):57-61.

PMID:14989932

Abstract

OBJECTIVE

To study whether macrophage migration inhibitory factor (MIF) can increase the ability of invasion of nasopharyngeal carcinoma cell lines in vitro, and to investigate the mechanism of invasion and metastasis of tumor cells during the early stage of nasopharyngeal carcinoma (NPC).

METHODS

The invasion and migration of NPC cell lines, CNE-1 and CNE-2, were evaluated by micron-migration assay in a chamber with 8- micro m porosity polycarbonate filter membrane. Flow cytometry and western blotting were adopted respectively to evaluate the protein expression level of matrix metalloproteinase 2 and 9 (MMP2, MMP9) in MIF treated or non-treated tumor cell lines. The concentrations of interleukin 8 (IL-8) secreted into the culture supernatant by the cells were measured by using Enzyme-linked immunoabsorbent assay (ELISA).

RESULTS

(1) After treatment with MIF for 24 hours, the number of cells passing through the 8- micro m filter membrane were increased in CNE-1 (113.7 +/- 20.9) and CNE-2 (311.3 +/- 48.9), as compared with that of non-MIF treated NPC cells. A significant statistic difference (P = 0.005, P = 0.001) was obtained in both CNE-1 and CNE-2 cells. (2) After treatment with MIF, the number of MMP9-positive cells increased in both CNE-1 (from 28.5% +/- 2.45% to 82.4% +/- 3.49%, P = 0.001) and CNE-2 (from 32.8% +/- 3.48% to 86.1% +/- 1.62%, P = 0.002) cell lines. In addition, an enhanced MMP9 protein expression up to 3-fold was observed in both cell lines. However, the expression level of MMP2 did not changed significantly between treated and non-treated cell lines (P > 0.05). (3) The concentration of IL-8 in the culture supernatant of CNE-2 was 1201.8 +/- 593.3 pg/ml after treatment with MIF for 24 h, remarkably higher than that without MIF treatment (32.7 +/- 20.1 pg/ml, P = 0.026). A similar change was not detected in CNE-1 (P = 0.581) cells.

CONCLUSIONS

(1) MIF can increase cell migration of CNE-1 and CNE-2 NPC cell lines in vitro. (2) A higher expression level of MMP9 and an up-regulated IL-8 by MIF may play a very important role in the progress of NPC, such as invasion and metastasis.

摘要

目的

研究巨噬细胞移动抑制因子（MIF）能否增强鼻咽癌细胞系的体外侵袭能力，并探讨鼻咽癌（NPC）早期肿瘤细胞侵袭和转移的机制。

方法

采用微孔迁移实验，通过8微米孔径的聚碳酸酯滤膜小室评估NPC细胞系CNE-1和CNE-2的侵袭和迁移能力。分别采用流式细胞术和蛋白质印迹法评估MIF处理或未处理的肿瘤细胞系中基质金属蛋白酶2和9（MMP2、MMP9）的蛋白表达水平。采用酶联免疫吸附测定（ELISA）法检测细胞分泌到培养上清液中的白细胞介素8（IL-8）浓度。

结果

（1）用MIF处理24小时后，与未用MIF处理的NPC细胞相比，CNE-1（113.7±20.9）和CNE-2（311.3±48.9）穿过8微米滤膜的细胞数量增加。CNE-1和CNE-2细胞均获得显著统计学差异（P = 0.005，P = 0.001）。（2）用MIF处理后，CNE-1（从28.5%±2.45%增至82.4%±3.49%，P = 0.001）和CNE-2（从32.8%±3.48%增至86.1%±1.62%，P = 0.002）细胞系中MMP9阳性细胞数量均增加。此外，在两个细胞系中均观察到MMP9蛋白表达增强至3倍。然而，处理和未处理的细胞系之间MMP2的表达水平没有显著变化（P>0.05）。（3）用MIF处理24小时后，CNE-2培养上清液中IL-8浓度为1201.8±593.3 pg/ml，显著高于未用MIF处理时（32.7±20.1 pg/ml，P = 0.026）。在CNE-1细胞中未检测到类似变化（P = 0.581）。