摘要
目的 研究巨噬细胞移动抑制因子 (MIF)体外提高鼻咽癌细胞侵袭能力的原因 ,探讨鼻咽癌细胞早期发生侵袭和转移的机制。方法 采用微孔迁移分析法 (micron migrationassay)检测鼻咽癌细胞株CNE 1、CNE 2在细胞因子MIF诱导下通过 8μm直径微孔的细胞数量变化 ,采用Western蛋白印迹法、流式细胞术和酶联免疫吸附法检测侵袭转移相关因子基质金属蛋白酶 2、9(MMP2 ,MMP9)及白介素 8(IL 8)在此过程中的相应改变。结果 (1)经MIF诱导后 ,CNE 1通过 8μm微孔的细胞数由原来的 2 3 7± 11 0 2增加至 113 7± 2 0 9,CNE 2则由原来的 3 4 7± 7 41增加至 3 11 3± 48 9,差异均呈显著性 (P =0 0 0 5,P =0 0 0 1)。 (2 )经MIF诱导后 ,癌细胞表达MMP9蛋白的细胞比例均明显增加 ,CNE 1细胞由 2 8 5%± 2 45%增加至 82 4%± 3 49% (P =0 0 0 1) ,而CNE 2细胞则由刺激前的 3 2 8%± 3 48%增加至 86 1%± 1 62 % (P =0 0 0 2 )。同时癌细胞MMP9蛋白表达强度也显著增强 ,CNE 1和CNE 2细胞MMP9蛋白的平均相对强度均比诱导前提高 3倍以上 (P <0 0 5)。但MIF刺激前后 ,MMP2蛋白的细胞表达数量和表达强度在两株细胞中却未见明显变化 (P值均大于 0 0 5)。 (3 )MIF诱导后 ,CNE 2细胞培养上清液中的IL 8浓度为
ObjectiveTo 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-μm porosity polycarbonate filter membra ne. 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-μm filter membrane were increased in CNE-1 (113.7±20.9) and CNE-2 (311.3±48.9), as compared with tha t 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,
出处
《中华病理学杂志》
CAS
CSCD
北大核心
2004年第1期57-61,共5页
Chinese Journal of Pathology
基金
国家自然科学基金资助项目( 30 2 0 0 2 54)
