Tanshinone â…¡A Induces Human Ovarian Cancer A2780 Cell Lines Apoptosis And Its Mechanism

Objective:Observe whether Tanshinoneâ…¡A can introduce apoptosis in human ovarian cancer A2780.Then explore it's mechanism to provide a theoretical basis to the clinical treatment of ovarian cancer.Methods:MTT assay was used to analysis the Proliferation of Tanshinoneâ…¡A to the A2780 cell line.Cell growth inhibitory rate=(1-0D values_{experimental groups}/OD value_{control group})×100%.The experiments were repeated three times.The results were from the average of the sum of the experiments' records.Cell growth inhibiting curve was drew by excel software with the horizontal axis and longitudinal axis representing concentration of Tanshinoneâ…¡A and cell growth inhibition ratio respectively.Half of the inhibitory rate(IC₅₀) was calculated by the software for IC₅₀ calculation. The cell apoptosis was determined by Flow cytometry.The experiments were divided into two groups:control group and experimental group.4.0 mg/L Tanâ…¡A was added into each group. According to the length of the time,we divide the experimental group into 24h group,48h group,and 72h group.Each group set up three parallel samples,1×10⁵ cells were inoculated into culture flask of 25 cm².Collecting the treated cells of each group,then fixed and stained.At last,cell apoptosis was detected by flow cytometryWestern blotting were applied to detect the expression of Bcl-2 and Bax in control group and the experimental groups.All the performance was carried out according to the kit instruction.Results:1.Tanâ…¡A could introduce apoptosis in human ovarian cancer A2780,and the intensity increased with time and the concentration of drug effects,acted as a time-dose-dependent manner.After treated with Tanâ…¡A for 48h,the IC₅₀ was 7.78 mg/L.2.FCM showed that after treated by Tanâ…¡A(4.0 mg/L),the apoptosis of A2780 ovarian cancer cells markedly increased. Quantitative FCM show the apotosis,A2780 cell line treated by 4.0 mg/L Tanâ…¡A for 0,24,48,72h,the apoptosis rates were 4.52%,19.7%, 25.3%,40.40%.Much more time.much more apoptotic cells.After treated for 72h,the apoptotic rate reached the highest value. Compared with the control group,24,48,72h drug treated group increased apoptosis ratewere statistically significant(P＜0.01).3.Western blot detected the protein expression of Bax and Bcl-2 in each grops,the results showed the expression of Bax could increase in the experimental group,but the expression of Bcl-2 declined.The ratio of Bcl-2/Bax had decreased markedly.Conlusion:1.Tanâ…¡A can significantly inhibit the proliferation of human ovarian cancer A2780 cells and induce the cells apoptosis.2.Declining the expression of Bcl-2 and increasing the expression of Bax might be the mechanism that Tanâ…¡A induced apoptosis in ovarian cancer A2780.