Effect of Trichostatin A on Apoptosis of Human Hepatoma Cell Line HepG2 and Expression of FHIT and Apoptosis-related Proteins
Duan Cheng-gang
Abstract
Duan Cheng-gang
Abstract
OBJECTIVE: To investigate the effect of trichostatin A (TSA),a specific inhibitor of histone acetyltransferase,on apoptosis of human hepotoma cell line HepG2,the expression of fragile histidine triad (FHIT) and apoptosis-related proteins in order to study apoptosis mechanisms.METHODS: Human hepatoma cell lines HepG2 were cultured and treated with different concentrations of TSA(125,250,500,1 000,2 000 nmol·L-1) for 24 and 48 hours.Human hepatoma cell lines HepG2 survival and apoptosis were determined by MTT assay with absorbance vale and inhibitory rate as index.Apoptotic percentage of HepG2 treated with 250 and 1 000 nmol·L-1 TSA were determined using TUNEL assay.The expressions of FHIT and caspase-3,bax,bcl-2 were analyzed by immunocytochemistry with solvent as control.RESULTS: As compared with control group,absorbance vale of human hepatoma cell lines HepG2 were decreased after treated with different concentration of TSA in dose-dependent and time-dependent manners.Positive cell rate in TUNEL was increased (P0.01),the protein expressions of FHIT and caspase-3,bax were up-regulated (P0.05) but the protein expression of bcl-2 had no obvious change (P0.05).CONCLUSION: TSA may inhibit proliferation and promote apoptosis of hepatoma HepG2 cell by up-regulating the protein expression of FHIT,caspase-3 and bax.
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OBJECTIVE: To investigate the effect of trichostatin A (TSA),a specific inhibitor of histone acetyltransferase,on apoptosis of human hepotoma cell line HepG2,the expression of fragile histidine triad (FHIT) and apoptosis-related proteins in order to study apoptosis mechanisms.METHODS: Human hepatoma cell lines HepG2 were cultured and treated with different concentrations of TSA(125,250,500,1 000,2 000 nmol·L-1) for 24 and 48 hours.Human hepatoma cell lines HepG2 survival and apoptosis were determined by MTT assay with absorbance vale and inhibitory rate as index.Apoptotic percentage of HepG2 treated with 250 and 1 000 nmol·L-1 TSA were determined using TUNEL assay.The expressions of FHIT and caspase-3,bax,bcl-2 were analyzed by immunocytochemistry with solvent as control.RESULTS: As compared with control group,absorbance vale of human hepatoma cell lines HepG2 were decreased after treated with different concentration of TSA in dose-dependent and time-dependent manners.Positive cell rate in TUNEL was increased (P0.01),the protein expressions of FHIT and caspase-3,bax were up-regulated (P0.05) but the protein expression of bcl-2 had no obvious change (P0.05).CONCLUSION: TSA may inhibit proliferation and promote apoptosis of hepatoma HepG2 cell by up-regulating the protein expression of FHIT,caspase-3 and bax.
Key concepts: Apoptosis, Trichostatin A, Molecular biology, FHIT, Cell culture, TUNEL assay, Chemistry, Immunocytochemistry