在人恶性胶质瘤原代培养细胞甲酰化肽受体FPR的表达和功能意义

Expression of functional formylpeptide receptor FPR by primary human malignant glioma cells

目的观测甲酰化肽受体(formylpeptidereceptor,FPR)在人恶性胶质瘤原代培养瘤细胞的表达和功能活性。方法组织块培养和胰酶消化法分离、培养人恶性胶质瘤细胞并进行鉴定,分别采用间接免疫荧光染色结合激光共聚焦扫描、台盼蓝排染实验结合细胞计数、酶联免疫夹心吸附分析(ELISA)等方法观测培养细胞FPR的表达和定位、FPR配体fMLF活化FPR后细胞增殖以及分泌血管内皮生长因子(vascularendothelialgrowthfactor,VEGF)的变化。结果部分胶质瘤原代培养细胞可见FPR表达,主要呈线性荧光信号定位于细胞膜;FPR阳性细胞对fMLF的刺激具有反应性,表现为细胞增殖速率加快,VEGF分泌量显著增加。结论部分人恶性胶质瘤细胞存在功能性FPR表达,并可促进瘤细胞增殖和VEGF的产生,在胶质瘤发展及血管生成过程中可能起重要作用。

Objective To detect the expression of formylpeptide receptor FPR in primary human glioma cells and its functional activity when stimulated with the ligand fMLF. Methods Human glioma tissues were employed with or without digestion with trypsin for primary culture of glioma cells. By using indirect immunofluorescence and confocal laser scanning microscopy, the expression and localization of FPR in the primary glioma cells were detected. Meanwhile, the cell proliferation in vitro and the production of vascular endothelial growth factor （VEGF） by primary cultured cells after FPR activation with its ligand fMLF, were assessed by trypan blue exclusion assay and enzyme-linked immunosorbent assay （ELISA） , respectively. Results The primary human malignant glioma cells expressed FPR on the plasma membrane, presenting discontinuous linear fluorescence. After FPR activation by fMLF, the cell proliferation in vitro was promoted, and the VEGF level secreted by primary glioma cells was increased significantly. A significantly positive correlation was found between FPR expression and the production of VEGF. Conclusion FPR was selectively expressed by the primary malignant glioma cells, and FPR activation promoted cell proliferation and the production of angiogenic factor VEGF in vitro.