[Molecular mechanisms of the protection of SNMC in HepG2 cell apoptosis].
OBJECTIVE
Apoptosis of the cells of liver cancer cell line HepG2 could be induced by TNF alpha and actinomycin D (Act D). In the current study, the molecular mechanism of the apoptosis protection of stronger neo-minophagen C (SNMC) to HepG2 cells was investigated.


METHODS
SNMC was added to the HepG2 cell culture medium when the cell concentration reached 0, 2, 20, 100, 200, 800 microg/ml 30 min before their apoptosis were inducted with TNF alpha and Act D. A flow cytometry assay was performed to detect the cell apoptosis rate; electromicroscopy was employed to visualize the subcellular structure after apoptosis. DNA ladder formation was checked with genomic DNA agarose electrophoresis. The expression pattern of apoptosis related protein Caspase-3, Bcl-2 and Bax was detected by Western blot.


RESULTS
After pretreatment with various concentrations of SNMC and 12 hours after treatment with TNF alpha and Act D, the HepG2 cell apoptosis rate and DNA ladder formation decreased dramatically when the SNMC concentration was higher in the media; the intracellular inactive form of Caspase-3 increased while the 17*10(3) active Caspase-3 decreased gradually. In addition, the expression of Bcl-2 increased and the expression of Bax decreased. Under the electromicroscope, the typical nucleolus condensation of HepG2 induced by TNF alpha and Act D was not seen among the 100 microg/ml SNMC treated cells.


CONCLUSIONS
SNMC inhibits TNF alpha and Act D induced HepG2 cell apoptosis. This protective action may be regulated by intracellular apoptosis related factors.
Wang
Yan
Y
Department of Infectious Diseases, First Affiliated Hospital, Harbin Medical University, Harbin 150001, China.
Ma
Ying-Ji
YJ
Yang
Bao-Shan
BS
Bi
Man-Ru
MR
Chen
Li-Yan
LY
D017209:Apoptosis / Q000187:drug effects*; D006528:Carcinoma, Hepatocellular / Q000473:pathology*; D045744:Cell Line, Tumor; D003545:Cysteine / Q000494:pharmacology*; D004338:Drug Combinations; D005998:Glycine / Q000494:pharmacology*; D006035:Glycyrrhiza; D006801:Humans; D008113:Liver Neoplasms / Q000473:pathology*; D009828:Oleanolic Acid / Q000031:analogs & derivatives* / Q000494:pharmacology
D004338:Drug Combinations; D009828:Oleanolic Acid; C009781:neominophagen C; D003545:Cysteine; D005998:Glycine