Bupleurum Injection On Human Hepatoma Cells In Hepg <sub> 2 </ Sub> Proliferation And Apoptosis In Impact

This article bases on lots of ancient Chinese Medicine classics,and makes an important summary and research about the history of literature and contemporary Doctors' Medical Books which related to liver cancer.This article also makes a comprehensive analysis about their academic development from the evolution of disease name,pathogenesis,and other aspects.Based on the relationship between Bupleurum injection and the major treatment about liver cancer,this article proposes the view that Bupleurum injection can inhibit liver cancer cell proliferation and promote liver cancer cell apoptosis.Objective:The way of this research is putting Bupleurum injection into invitro human liver cancer cell line HepG₂,in order to inhibit their proliferation,apoptosis,and explore the possible mechanism from the molecular level.Methods:1 The experimental research about Bupleurum injection inhibit the proliferation of liver cancer cell line HepG₂.1.1 control group(10%calf serum containing 1640 medium)and 5-fluorouracil positive control group(peak blood concentration of 10μg / L),and Bupleurum Injection 3 mg / ml,5mg/ml,10mg/ml,25mg / ml,50mg/ml different concentrations of the experimental group.1.2 Test the sensitivity of liver cancer cell line HepG₂ by blue assay(MTT)on Bupleurum injection and 5-fluorouracil.1.3 Apply flow cytometry(FCM)detect the cycle change of liver cancer cell line HepG₂.2 The research of Bupleurum injection due to HepG₂ cells apoptosis.2.1 Apply inverted microscope fluorescence microscopy to observe the changes of apoptosis. 2.2 Apply FCM to detect the apoptosis rate.2.3 One-step TUNEL detect apoptosis rate.Results:1 Bupleurum injection can significantly inhibit the growth of HepG₂ and make an induction of apoptosis on the HepG₂.The MTT test showed that Bupleurum injection could make human liver cancer cell proliferate in a dose-effect relationship——the more concentration,the more incidence of inhibition.Its 50%inhibitory concentration (IC₅₀)is about 50 mg / ml.There are no significant difference between Bupleurum injection micro-dose(3 mg/ ml)and the control group and there are also no significant diversity between Bupleurum injection 25 mg / ml group and the positive control group 5-fluorouracil. However the difference between compared and the other two groups are obviously. According to FCM results that separately using the concentration of 5 mg / ml, 10mg/ml and 25mg/ml Bupleurum injection to human liver cancer cells HepG₂ 12 hours,the percentage of the cell in the period of G₁ are continuing decreased.And the G₁%of concentration about 5 mg / ml,10mg/ml,25mg/ml,G₁%are respectively 66.30%and 57.27%,49.23%.This shows that the main inhibition of Bupleurum injection on human liver cancer cells HepG₂ is in G₁ phase.2 The effect of Bupleurum injection on vitro liver cancer cells HepG₂2.1 After 12 hours injecting Bupleurum injection,compared with the control group, HepG₂ cells form changed significantly.The inverted microscope observed that the volume of cell became narrow,round,and the volume of the cytoplasm decreased. With the increased of concentration,the cell wall gradually reduced,suspended cells increased.When the concentration of Bupleurum injection reached to 25 mg / ml in experimental group,the effects are hardly similar with the 5-fluorouraci control group.2.2 The HepG₂ cells could be emphasized by Bupleurum injection for the histogram hypodiploid peak could be seen on the flow cytomtry.And theapoptosis rate increased with the concentration of drug increased——dose-dependence. 2.3 One-step TUNEL test show that after using Bupleurum injection 12 hours,the apoptosis rate was significantly higher than the control group.Conclusion1 Bupleurum injection can inhibit HepG₂ cell proliferation,HepG₂ cells are inhibited in G₁ phase.2 Bupleurum injection can due to HepG₂ apoptosis with apoptosis-specific morphological and biochemical changes.3 The active ingredients in Bupleurum injection may be the potential drug on the treatment of liver cancer.