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


CONCLUSIONS
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.
Yao
Xiao-hong
XH
Institute of Pathology, Southwest Hospital, Third Military Medical University, Chongqing 400038, China.
Ping
Yi-fang
YF
Bian
Xiu-wu
XW
Chen
Jian-hong
JH
Wang
Qing-liang
QL
Jiang
Xue-feng
XF
D045744:Cell Line, Tumor; D049109:Cell Proliferation / Q000187:drug effects*; D002470:Cell Survival / Q000187:drug effects; D004797:Enzyme-Linked Immunosorbent Assay; D005910:Glioma / Q000235:genetics / Q000378:metabolism / Q000473:pathology; D006801:Humans; D016209:Interleukin-8 / Q000235:genetics / Q000378:metabolism*; D009637:Masoprocol / Q000031:analogs & derivatives* / Q000494:pharmacology; D018613:Microscopy, Confocal; D009240:N-Formylmethionine Leucyl-Phenylalanine / Q000494:pharmacology; D012333:RNA, Messenger / Q000235:genetics / Q000378:metabolism; D044042:Receptors, Formyl Peptide / Q000096:biosynthesis*; D020133:Reverse Transcriptase Polymerase Chain Reaction; D042461:Vascular Endothelial Growth Factor A / Q000235:genetics / Q000378:metabolism*
D016209:Interleukin-8; D012333:RNA, Messenger; D044042:Receptors, Formyl Peptide; C467484:VEGFA protein, human; D042461:Vascular Endothelial Growth Factor A; C510003:nordy; D009240:N-Formylmethionine Leucyl-Phenylalanine; D009637:Masoprocol