[诺帝抑制甲酰肽受体介导的人恶性胶质瘤细胞增殖及血管生成因子产生]

[Nordy inhibits cell proliferation and angiogenic factor production of malignant human glioma cells mediated by formylpeptide receptor].

作者信息

Yao Xiao-hong, Ping Yi-fang, Bian Xiu-wu, Chen Jian-hong, Wang Qing-liang, Jiang Xue-feng

机构信息

Institute of Pathology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.

出版信息

Zhonghua Yi Xue Za Zhi. 2007 Feb 13;87(7):479-84.

PMID:17459229

Abstract

OBJECTIVE

To explore the effect of Nordy, a synthesized lipoxygenase Inhibitor, on the expression of formylpeptide receptor (FPR) and its functional activities after activation in human malignant glioma cells.

METHODS

Human malignant glioma cells of the line U87 were cultured and divided into 3 groups: N-formyl-methionyl-leucyl-phenylalanine (fMLF) group, added with 100 nmol/L fMLF; Nordy group, added with 100 micromol/L Nordy for 12 h and then added with 100 nmol/L fMLF; and control group. The expression of FPR I the U87 cells was detected with indirect immunofluorescence staining and confocal laser scanning microscopy. The effect of Nordy on the U87 cell proliferation induced by fMLF was assayed by MTT method. RT-PCR was used to detect the mRNA expression of vascular endothelial growth factor (VEGF) and interleukin-8 (IL-8) in the U87 cells. ELISA was used to detect protein secretion of VEGF and IL-8 in the supernatant.

RESULTS

FPR was expressed on the cytoplasmic membrane of U87 cells. The mean FPR absorbance of the Nordy group was significantly lower than that of the fMLF group. The U87 cell proliferation of the Nordy group was significantly inhibited in comparison with the other 2 groups. The mRNA expression of VEGF and mRNA expression of IL-8 in the U87 cells activated by fMLF were significantly higher than those of the control group (both P<0.05), and Nordy significantly attenuated the mRNA expression of VEGF and IL-8 at different time points (all P<0.05). ELISA showed that the VEGF and IL-8 protein levels in the supernatant were 4.14 ng/ml+/-0.28 ng/ml and 4.03 ng/ml+/-0.59 ng/ml respectively, and increased to 6.46 ng/ml+/-0.33 ng/ml and 7.54 ng/ml+/-0.73 ng/ml respectively after stimulation of fMLF, however, the VEGF and IL-8 protein levels in the supernatant of the Nordy group were 3.59 ng/ml+/-0.33 ng/ml and 3.13 ng/ml+/-0.48 ng/ml respectively.

CONCLUSION

Nordy inhibits the FPR expression and the effects in promoting the cell proliferation and mRNA expression and protein secretion of VEGF and IL-8 after the activation of FPR in malignant human glioma cells, showing that Nordy has the effects of inhibition of tumor growth and angiogenesis.

摘要

目的

探讨合成脂氧合酶抑制剂诺帝（Nordy）对人恶性胶质瘤细胞中甲酸肽受体（FPR）表达及其激活后功能活性的影响。

方法

培养人恶性胶质瘤U87细胞系，分为3组：N-甲酰甲硫氨酰-亮氨酰-苯丙氨酸（fMLF）组，加入100 nmol/L fMLF；诺帝组，先加入100 μmol/L诺帝作用12 h，再加入100 nmol/L fMLF；对照组。采用间接免疫荧光染色和共聚焦激光扫描显微镜检测U87细胞中FPR的表达。采用MTT法检测诺帝对fMLF诱导的U87细胞增殖的影响。采用逆转录-聚合酶链反应（RT-PCR）检测U87细胞中血管内皮生长因子（VEGF）和白细胞介素-8（IL-8）的mRNA表达。采用酶联免疫吸附测定（ELISA）检测上清液中VEGF和IL-8的蛋白分泌。

结果

FPR表达于U87细胞的细胞膜上。诺帝组FPR平均吸光度显著低于fMLF组。与其他2组相比，诺帝组U87细胞增殖受到显著抑制。fMLF激活的U87细胞中VEGF的mRNA表达和IL-8的mRNA表达均显著高于对照组（均P<0.05），诺帝在不同时间点均显著减弱VEGF和IL-8的mRNA表达（均P<0.05）。ELISA结果显示，上清液中VEGF和IL-8蛋白水平分别为4.14 ng/ml±0.28 ng/ml和4.03 ng/ml±0.59 ng/ml，fMLF刺激后分别升高至6.46 ng/ml±0.33 ng/ml和7.54 ng/ml±0.73 ng/ml，而诺帝组上清液中VEGF和IL-8蛋白水平分别为3.59 ng/ml±0.33 ng/ml和3.13 ng/ml±0.48 ng/ml。