Inhibition effect of MIF antibody on the growth of hepatocellular carcinoma cell HepG2 in vitro.
- Author:
Hong PENG
1
;
Zu-Kui YANG
;
Jun HOU
Author Information
- Publication Type:Journal Article
- MeSH: Antibodies; pharmacology; Cell Cycle; Cell Proliferation; drug effects; Cyclin D1; metabolism; Hep G2 Cells; Humans; Interleukin-6; metabolism; Macrophage Migration-Inhibitory Factors; immunology; Vascular Endothelial Growth Factor A; metabolism
- From: Chinese Journal of Hepatology 2008;16(12):918-921
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVESTo investigate the inhibitory effect of macrophage migration inhibitory factor (MIF) antibody on the proliferation of HepG-2 cells and its mechanism.
METHODSHepG-2 cells were stimulated by different concentrations of MIF antibody (50, 100, 200 and 400 microg/L). The cell survival rates were evaluated by MTT assay. The cell cycles were assessed by flow cytometry (FCM) analysis. Cyclin D1 protein expression was examined by immunohistochemical methods. Vascular endothelial growth factor (VEGF) protein expression was examined by Western blot. ELISA was applied to detect the influence of MIF antibody on the production of IL-6 of HepG-2 cells.
RESULTSHepG-2 cells were inhibited by MIF antibody in a dose and time dependent manner. FCM analysis showed the cell cycles of HepG-2 cells were blocked at G0/G1 phase. With concentrations of MIF antibody of 0, 50, 100, 200, 400 microg/L, the percentages of cells in G0/G1 phase at 48 h were 61.34%+/-1.08%, 65.08%+/-2.71%, 71.19%+/-1.19%, 78.39%+/-1.00%, 83.92%+/-0.51%. With concentrations of MIF antibody of 50, 100, 200, 400 microg/L, the expressions of cyclin D1 protein were 26.06%+/-0.47%, 22.34%+/-0.75%, 18.06%+/-1.16%, 14.03%+/-0.59%, significantly lower than those of the control group (29.51%+/-1.28%). When HepG-2 cells were treated with different concentrations of MIF antibodies of 50, 100, 200, 400 microg/L the expressions of VEGF protein were 21.22%+/-0.68%, 19.64%+/-0.54%, 18.04%+/-0.42%, 16.59%+/-0.66%, significantly lower than those of the control group (23.23%+/-0.51%). With MIF antibody concentrations of 0, 50, 100, 200, 400 microg/L, the exudation amount of IL-6 were correspondingly lower [(210.67+/-9.31) pg/ml, (181.67+/-10.05) pg/ml, (160.50+/-6.60) pg/ml, (143.67+/-10.56) pg/ml, (118.01+/-7.48) pg/m].
CONCLUSIONThe proliferation of HepG-2 cells was inhibited after treatment with MIF antibody. The inhibiting effect is caused by blocking cell cycle progression at G0/G1 phase, decreasing cyclin D1 protein expression, decreasing VEGF protein expression and decreasing the exudation amount of IL-6.
