Formylpeptide receptor FPR and the rapid growth of malignant human gliomas
Formylpeptide receptor FPR and the rapid growth of malignant human gliomas
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DOI：
10.1093/jnci/dji142
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发表时间：
2005-06-01
影响因子：
10.3
通讯作者：
Wang, JM
中科院分区：
文献类型：
--
作者：
Zhou, Y;Bian, XW;Wang, JM
背景甲酰基肽受体（FPR）是一种G蛋白偶联受体（GPCR），介导细菌肽N-甲酰基-甲硫氨酰-亮氨酰-苯丙氨酸（fMLF）诱导的吞噬白细胞趋化性。我们先前表明，选定的人胶质瘤细胞系也表达功能性FPR。因此，我们研究了FPR表达与胶质瘤细胞生物学行为之间的关系。研究方法：应用逆转录-聚合酶链反应（RT-PCR）和趋化性实验检测FPR在人胶质母细胞瘤细胞系U-87中的表达和功能。应用免疫组化方法检测33例人脑胶质瘤中FPR蛋白的表达。使用FPR短干扰（si）RNA来阻断U-87细胞中FPR的表达。通过测量DNA合成来评估细胞增殖。在裸鼠中测量异种移植肿瘤形成和生长。通过测量FPR转染的嗜碱性粒细胞白血病细胞系和活U-87细胞中的FPR活化来评估坏死肿瘤细胞释放的内源性FPR激动剂活性。RT-PCR检测血管内皮生长因子（VEGF）mRNA表达，酶联免疫吸附试验检测VEGF蛋白表达。所有统计学检验均为双侧检验。结果如下：FPR在高度恶性的人胶质母细胞瘤细胞系U-87和大多数原发性IV级多形性胶质母细胞瘤和III级间变性星形细胞瘤中选择性表达。U-87细胞通过趋化性（即，增加的运动性）、增加的细胞增殖和增加的VEGF蛋白产生。FPR SiRNA显著降低U-87细胞在裸小鼠体内的致瘤性（植入后38天，野生型U-87细胞的平均肿瘤体积= 842 mm 3（3），95%置信区间[CI] = 721至963 mm 3（3）; FPR-siRNA转染的U-87细胞的平均肿瘤体积= 225 mm 3，95% CI = 194至256 mm 3（3）; P =.001）。坏死的胶质母细胞瘤细胞释放激活活U-87细胞中FPR的因子。结论：FPR由高度恶性的人胶质瘤细胞表达，并且似乎通过与宿主衍生的激动剂相互作用来介导人胶质母细胞瘤的运动性、生长和血管生成。因此，FPR可能是开发新型抗胶质瘤治疗药物的分子靶点。
Background. The formylpeptide receptor (FPR) is a G-protein-coupled receptor (GPCR) that mediates chemotaxis of phagocytic leukocytes induced by bacterial peptide N-formyl-methionyl-leucyl-phenylalanine (fMLF). We previously showed that selected human glioma cell lines also express functional FPR. We therefore investigated the relationship between FPR expression and the biologic behavior of glioma cells. Methods: Expression and function of FPR in the human glioblastoma cell line U-87 were examined by reverse transcription-polymerase chain reaction (RT-PCR) and chemotaxis assays, respectively. FPR protein expression was detected in specimens from 33 human primary gliomas by immunohistochemistry. FPR short interfering (si) RNA was used to block FPR expression in U-87 cells. Cell proliferation was assessed by measuring DNA synthesis. Xenograft tumor formation and growth were measured in nude mice. Endogenous FPR agonist activity released by necrotic tumor cells was assessed by measuring FPR activation in an FPR-transfected basophil leukemia cell line and live U-87 cells. Vascular endothelial growth factor (VEGF) mRNA was assessed by RT-PCR, and VEGF protein was assessed by enzyme-linked immunosorbent assay. All statistical tests were two-sided. Results: FPR was selectively expressed by the highly malignant human glioblastoma cell line U-87 and most primary grade IV glioblastomas multiforme and grade III anaplastic astrocytomas. U-87 cells responded to the FPR agonist fMLF by chemotaxis (i.e., increased motility), increased cell proliferation, and increased production of VEGF protein. FPR siRNA substantially reduced the tumorigenicity of U-87 cells in nude mice (38 days after implantation, mean tumor volume from wild-type U-87 cells = 842 mm(3), 95% confidence interval [CI] = 721 to 963 mm(3); and from FPR-siRNA transfected U-87 cells = 225 mm3, 95% Cl = 194 to 256 mm(3); P =.001). Necrotic glioblastoma cells released a factor(s) that activated FPR in live U-87 cells. Conclusions: FPR is expressed by highly malignant human glioma cells and appears to mediate motility, growth, and angiogenesis of human glioblastoma by interacting with host-derived agonists. Thus, FPR may represent a molecular target for the development of novel antiglioma therapeutics.