Effects of exogenous epidermal growth factor (EGF) on growth inhibition induced by aristolochic acid I (AA-I) in renal proximal tubular epithelial cells.
- Author:
Na ZHOU
1
;
Li YANG
;
Jia-Wei TANG
;
Xiao-Mei LI
Author Information
- Publication Type:Journal Article
- MeSH: Aristolochic Acids; pharmacology; Cell Cycle; drug effects; Cell Line; Cell Proliferation; drug effects; Epidermal Growth Factor; pharmacology; Epithelial Cells; drug effects; Flow Cytometry; Humans; Kidney Tubules, Proximal; cytology; drug effects; Mutagens; pharmacology
- From: China Journal of Chinese Materia Medica 2008;33(19):2222-2226
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVEThe purpose of this study was to investigate the effects of exogenous epidermal growth factor (EGF) on the growth inhibition of renal proximal tubular epithelial cell induced by AA-I.
METHODCultured human renal proximal tubular epithelial cell line HK-2 was used as the subject. The changes of the survived HK-2 cells were observed and compared among control, AA-I stimulation, pre-EGF, together-EGF and post-EGF groups. In the study, cellular morphologic assessments were performed with a phase-contrast inverted microscope. Cell counting after stained with 0.04% trypan blue was adopted to analyze cell proliferation. Cell cycle was assessed by flow cytometry.
RESULTNumber of the survived cells stimulated by AA-I for 12, 24, 48 hours decreased gradually in a dose and time dependent manner. At 24, 48 hours, the survived cells showed a significant disturbance in the cell cycle procedure, which was characterized as decreased percentage of cells in G0/G1 phase, significant increased percentage of cells in G2/M phases. Exogenous EGF (20 microg L(-1)) could significantly promote the proliferation of HK-2 cells, which shown a increased cell number, accompanied down-regulated cells in G0/G1 phase, increased cells in S and G2/M phase. Compared with AA-I groups, it failed to improve the inhibitory effect on cell proliferation and abnormal cell cycle procedure by AA-I, no matter EGF was added in before, at same time or after AA-I stimulation.
CONCLUSIONAA-I (10 g L(-1)) has remarkable growth inhibition effects on survived RTEC, and can induce a blockage of G2/M phase in the cell cycle procedure. Exogenous EGF (20 microg L(-1)) promote the proliferation in normal cultured HK-2 cell. EGF treatment could not improve the proliferation inhibitory effect induced by AA-I, no matter adding EGF to cultures before, together with AA-I or after AA-I stimulation.