Effect of Tanshinone II A on expression of EGF and EGFR in hepatocellular carcinoma cell line SMMC-7721.
- Author:
Xue-min ZHAI
1
;
Shui-xiang HE
;
Mu-dan REN
;
Jing-hong CHEN
;
Zhi-lun WANG
;
Min HAN
;
He-lei HOU
Author Information
- Publication Type:Journal Article
- MeSH: Antineoplastic Agents, Phytogenic; pharmacology; Apoptosis; drug effects; Carcinoma, Hepatocellular; metabolism; pathology; Cell Line, Tumor; Cell Proliferation; Diterpenes, Abietane; Down-Regulation; drug effects; Epidermal Growth Factor; genetics; metabolism; Humans; Liver Neoplasms; metabolism; pathology; Phenanthrenes; pharmacology; Receptor, Epidermal Growth Factor; genetics; metabolism
- From: Journal of Zhejiang University. Medical sciences 2009;38(2):163-169
- CountryChina
- Language:Chinese
-
Abstract:
OBJECTIVETo observe the effect of Tanshinone II A on the expression of epidermal growth facter (EGF) and epidermal growth facter recepter (EGFR) in human hepatocellular carcinoma cell line SMMC-7721.
METHODSThe human hepatocellular carcinoma SMMC-7721 cells cultured in vitro was treated with different concentrations of Tanshinone II A. The proliferation of the cells was measured by MTT assay, and the apoptosis of the cells was investigated by flow cytometry and cytochemical staining with Hoechst 33342. The expression of EGF and EGFR was detected by immunocytochemistry method. The levels of EGF in medium were measured by radioimmunoassay.
RESULTTanshinone II A inhibited the growth of SMMC-7721 cells remarkably in a dose-dependent manner. The inhibitory rate reached the peak (72.5%) after 0.5 microg/ml Tanshinone II A was used for 48 h, which was significantly higher than that in the controls (P<0.05). FCM analysis showed that when SMMC-7721 cells were treated with 0.5 microg/ml Tanshinone II A, the apoptosis rates for 24 h, 48 h and 72 h were (4.06+/-0.27)%, (7.58+/-0.56)% and (5.23+/-0.13)%, respectively which were markedly higher than those in the controls (all P<0.01). SMMC-7721 cell apoptosis with cell shrinkage, nuclear chromatin concentration and fragmentation as well as the formation of apoptotic bodies were observed by cytochemical staining when treated with Tanshinone II A. The immunocytochemistry showed that the expressions of EGF and EGFR were down regulated while the concentration of Tanshinone II A was increasing. The high expression rates for EGF and EGFR were 10%, 20%, respectively, and the gray scale was 181.52+/-1.63, 179.37+/-1.59, which were markedly higher than those in the controls (all P<0.05). The levels of EGF in medium measured by radioimmunoassay were decreased significantly after Tanshinone II A treatment.
CONCLUSIONTanshinone II A can inhibit cell proliferation and induce apoptosis in hepatocellular carcinoma cell line SMMC-7721, which may be related to the down-regulation of EGF and EGFR protein expression.
