Han Anti A Induced By The Human Hepatoma Cell Line In Hepg <sub> 2 </ Sub> Apoptosis And Bax And Caspase-3

Objectives:In present study we effected human hepatocellular carcinoma cells HepG₂ by in vitro cultivated tetrandrine (Tet) , to restrain it' s proliferation and induce apoptosis, to investigate the mechanism on molecule level.Methods:1 Tet restrain HepG₂ cells proliferation . MTT was used to detect the medicine sensitivity of HepG₂ cells. And flow cytometry(FCM) was used to detect the cell cycle.2 Tet induce HepG₂ cells apoptosis. By means of invert microscope, microscope and electron microscope, the form changes of Tet on HepG₂ cells were investigated. FCM and TUNEL were performed to check the changes in apoptotic rate of HepG₂ cells.3 Expression of Bax and Caspase-3 proteins were investigated with immunohistochemical methods.Result:1 Tet restrain HepG₂ cells proliferation :1.1 Apoptosis was seen in HepG₂ cells treated with Tet, dose dependent effects of Tet to HepG₂ cells were detected by MTT. Half-maximal inhibtion of growth was observed at concentration of 9. 3ug/ml.1.2 Tet can increase the percentages of G, phases in HepG₂ cell cycles at 8.0 ug/ml; Tet also increase the percentages of G₂ phases in HepG₂cell cycles at 9. 3ug/mk 10. 5ug/ml.2 Tet induce HepG₂ cells apoptosis:2.1 After HepG₂ cells were treated with Tet for 24h, the typical changes of apoptosis of HepG₂ cells were found. Not only the volume of HepG₂ became small and round, but also the nuclear chromatine condensation and fragmentation were observed in HepG₂ cells.2.2 Hypodiploid peak appeared on FCM histogram, and the apoptotic rate of HepG₂ cells was risen with the dose.2. 3 After HepG₂ cells were treated with Tet for 24 h, the apoptotie rate was significantly different from those in blank control groups by TUNEL. (all P<0. 01)3 Tet could obviously increase the expression of Bax gene and Caspase-3 gene with immunohistochemical methods. It was possible pathway of Tet inducing apoptosis of HepG₂ cells.