Effects Of Casticin On Apoptosis And Differentiation Of Hepatocellular Carcinoma HepG2 Cells

Objective:to observe the effects of casticin(CAS) on the growth inhibition, differentiation and apoptosis of human hepatocellular carcinom Hep G2 cells in vitro, and evaluate the potential value of CAS as a novel agent for human liver cancer treatment.Methods:After exposure o hepatocellular carcinoma Hep G2 cell line to different concentrations of CAS, MTT assay, colony formation method and DNA agarose gel electrophoresis were used to evaluate the growth and apoptosis of cells. Flow cytometry (FCM) using PI staining was used to detemine cell cycle distribution and apoptosis. The caspase-3 Activity were detected by enzyme linked immunosorbent assay(ELISA). Microscope using Wright-Gimsa staining was used to measure the nuclear-cytoplasmic ratio. The secretion of alpha-fetoprotein (AFP) was assayed by radio immunoassay. The activity of alkaline phosphatase (ALP) andγ-glutamyltranspeptidase (γ-GT) were tested by enzymaticreaction kit. Diamond stone spectrophotometry was employed to determine the activity of tyrosine-α-Ketoglutaric acid transaminase (TAT).Results:After treatment of human hepatocellular carcinoma Hep G2 cell line with various concentrations of CAS, the cell proliferation and anchorage-dependent growth inhibition were observed, the difference was statistically significant compared with the control group (P<0.01).10 mg/L CAS decreased the percentage of G0/G1 phase cells (P< 0.01), and increased the percentage of S phase and G2/M phase cells(P< 0.01) Sub-G1 population significantly increased(P< 0.1). In CAS treated group, DNA of Hep G2 cells was significantly degraded, as showed by characteristic apoptotic DNA ladder bands. CAS increased the caspase-3 activity of HepG2 cells, the difference was statistically significant compared with the control group (P< 0.01); The activities of TAT and ALP in Hep G2 cells were significantly increased by CAS(P< 0.01), meanwhile the secretion of AFP and the activities ofγ-GT were decreased respectively(P< 0.01).Conclusion:1.CAS significantly inhibits the growth of human hepatocarcinoma Hep G2 cells.2. CAS induces the apoptosis of human hepatocarcinoma Hep G2 cells.3. CAS induces the differentiation of human hepatocarcinoma Hep G2 cells.4. Induction of the growth inhibition and apoptosis by CAS in human hepatocarcinoma Hep G2 cells may be associated with arrest of G2/M phase in cell cycle and activation of caspase-3.