恶性胶质瘤细胞甲酰化肽受体表达及其活化后促瘤细胞增殖和血管生成因子产生的作用  

Activation of Formylpeptide Receptor in Human Malignant Glioma Cells and Its Effects on Cell Proliferation and Production of Angiogenic Factors

作　　者：姚小红[1] 卞修武[1] 平轶芳[1] 陈剑鸿[2] 王清良[1] 蒋雪峰[1] 

机构地区：[1]第三军医大学西南医院病理学研究所,重庆400038 [2]第三军医大学西南医院药学部,重庆400038

出　　处：《癌症》2007年第3期241-246,共6页Chinese Journal of Cancer

基　　金：国家自然科学基金项目(No.30370552)~~

摘　　要：背景与目的:趋化因子受体在多种病理过程中发挥重要作用,通过促进肿瘤细胞增殖、迁移或血管生成过程,进而在肿瘤发生、演进和转移中发挥极为重要的作用。本研究拟探讨甲酰化肽受体(formylpeptide receptor,FPR)在人恶性胶质瘤细胞中的表达和激活后的功能活性。方法:以人恶性胶质瘤细胞U87和FPR-siRNA-U87细胞为研究对象,采用间接免疫荧光标记、激光共聚焦扫描显微术观测人恶性胶质瘤细胞FPR的表达和定位;用FPR配体甲酰-甲硫酰-亮氨酰-苯丙氨酰胺(formy-Met-Leu-Phe,fMLF)激活FPR后,采用四甲基偶氮唑蓝(MTT)法检测细胞增殖,双抗夹心酶联免疫(ELISA)吸附法检测细胞分泌血管内皮生长因子(vascular endothelial growth factor,VEGF)和白细胞介素8(interleukin-8,IL-8)水平,RT-PCR法检测细胞IL-8mRNA的水平。结果:恶性胶质瘤细胞U87有FPR表达,定位于细胞膜,FPR-siRNA-U87细胞不表达FPR;FPR激动剂fMLF对人恶性胶质瘤细胞具有明显的促增殖作用,U87细胞fMLF刺激组A值较对照组显著升高(P<0.05),FPR-siRNA-U87细胞fMLF刺激组A值与对照组相比无显著性差异(P>0.05);fMLF作为FPR的激动剂,在12h、24h、36h和48h等各时相点U87细胞的VEGF和IL-8蛋白水平均显著增加,以处理36h为观察时间点,U87细胞VEGF蛋白水平[(3.13±0.23)ng/ml]较对照组显著增加(P<0.05),IL-8蛋白分泌量为(7.54±0.53)ng/ml,较对照组显著增加(P<0.05);FPR-siRNA-U87细胞VEGF和IL-8蛋白分泌量分别为(1.26±0.05)ng/ml和(1.54±0.05)ng/ml,较对照组无明显改变(P>0.05);fMLF能显著上调U87细胞IL-8mRNA的表达,较对照组显著升高(P<0.05),FPR-siRNA-U87细胞IL-8mRNA水平与对照组相比无显著性差异(P>0.05)。结论:人恶性胶质瘤细胞膜存在功能性FPR受体,其活化后具有促瘤细胞增殖和分泌血管生成因子的作用。BACKGROUND ＆ OBJECTIVE： Chemoattractant receptors participate in essential pathophysiologic processes, including inflammation, wound healing, human immunodeficiency virus infection, and most interestingly in the progression of malignant tumor. This study was to explore the functional expression of formylpeptide receptor （FPR） in human glioblastoma cell line U87. METHODS. The expression of FPR in U87 and FPR small interfering RNA （siRNA）-transfected U87 cells （FPR-siRNA-U87 cells） was detected with indirect immunofluorescent staining by confocal laser scanning microscopy. FPR was activated by its ligand formy-Met-Leu- Phe （fMLF）. Cell proliferation was assessed by MTT assay. The production of vascular endothelial growth factor （VEGF） and interleukin-8 （IL-8） was measured by ELISA, and IL-8 mRNA was assessed by reverse transcriptionpolymerase chain reaction （RT-PCR）. RESULTS; FPR was expressed on U87 cells, but not on FPR-siRNA-U87 cells. After activation of FPR by fMLF, the proliferation of U87 cells was enhanced markedly （P〈0.05）, but that of FPR-siRNA-U87 cells had no obvious change （P〉0.05）. fMLF （100 nmol/L） elicited a time-dependent increase in the secretion of IL-8 and VEGF. When stimulated with 100 nmol/L fMLF for 36 h, the protein levels of VEGF and IL-8 were significantly higher in stimulated U87 cells than in control cells [（3.13± 0.23） ng/ml vs. （2.55±0.25） ng/ml, P〈0.05; （7.54±0.53） ng/ml vs. （4.02±0.09） ng/ml, P〈0.05], but those in FPR-siRNA-U87 cells had no obvious change; the mRNA level of IL-8 was significantly increased in U87 cells, but not changed in FPR-siRNA-U87 cells. CONCLUSION, Activating FPR can promote the proliferation of U87 cells and enhance the production of angiogenic factors.

关 键 词：甲酰化肽受体(FPR) 脑肿瘤 胶质瘤 血管生成因子 细胞增殖 血管内皮生长因子 白细胞介素8 

分 类 号：R739.4[医药卫生—肿瘤]