Matrine Induces Cell Cycle Arrest And Apoptosis In Gallbladder Carcinoma Cell Lines

Background:Gallbladder carcinoma is the fifth most common gastrointestinal malignancy and the most common biliary tract malignancy worldwide . This tumour is traditionally regarded as a highly lethal disease with an over all 5-year survival of less than 5%. Aggressive biological behaviour of the tumour and the late presentation of the disease contributing to the dismal prognosis.Because of the radioresistance of gallbladdercarcinorma cells and the difficulty of achieving complete tumor resection, the current treatments are ultimately ineffective. There is therefore an urgent need to seek novel therapeutic drugs to combat Gallbladder carcinoma.Matrine is the major active component of the traditional Chinese medicine Sophora flavescens, with a molecular formula of C₁₅H₂₄N₂₀. It has been widely used in China for the treatment of viral hepatitis, hepatic fibrosis, arrhythmia, atherosclerosis, and skin diseases. Recently, it has been reported that matrine exhibits anti-tumor effects by inhibiting proliferation and inducing apoptosis of cancer cells from cervical cancer, leukemia, gastric cancer, hepatocellular carcinoma, breast cancer, and lung cancer. However,there's no reserch reported about the matrine's effect on gallbladder carcinoma. In this study, we used GBC-SD cells to investigate whether matrine could induce apoptosis or inhibit the proliferation of gallbladder carcinoma cells in vitro, and the possible mechanism was also explored.Objectives:Matrine, the major active component of the traditional Chinese medicine Sophora flavescens, with a molecular formula of Cl₅H₂₄N₂₀, It has been widely used in China for the treatment of viral hepatitis, hepatic fibrosis, arrhythmia, atherosclerosis, and skin diseases. Current researches indicate that it may also be one of the effective tumor suppressive agents. Hence, this present study was carried out to investigate the proliferation inhibition and apoptosis induction effect of matrine on human gallbladder cancer GBC-SD cells and the possible mechanisms.Methods:The GBC-SD cells were treated with matrine at various concentrations combination of cisplatin in present study. The MTT assay was used to assess cytostatic activity induced by matrine. Flow cytometry were adopted to evaluate the cell cycle and the cell apoptosis after matrine treatment. Western blotting was used to demonstrate expression levels of involved protein such as cdks ,cyclins,bcl-2, bax, caspase-3.All the detection items in this study were repeated at least 3 times. Statistical analysis was done using SPSS software. The data was expressed as mean±SD and the statistical significance of the differences between control and matrine treated cells was determined by a two-tailed Student's t test. The Spearman correlation test was used to analyze the correlation between reagents' concentrations and inhibition rates. P-value <0.05 was considered as significant. Results:The MTT assay indicated that the GBC-SD cells proliferation could be dose-and time-dependently inhibited by matrine. Flow cytometry showed that matrine significantly induced cell cycle arrest and apoptosis in GBC-SD cell line .According to the results of Western Blot, bcl-2 and cyclinE were significantly downregulated in cells treated with matrine while bax was upregulated in cells treated with matrine. Moreover, the different effects expressions of caspase-3 was detected by Western blot analysis.Conclusions:In this study, the apoptosis induced effect of matrine on human gallbladder carcinorma GBC-SD cell line was analyzed. The matrine can inhibit growth, induce G1 cell cycle arrest ,also can induce apoptosis of GBC-SD cells. The findings presented here reveal that the G1 cycle arrest treated with matrine was associated with the down regulation of cyclinE, and matrine can induce marked apoptosis mainly influencing the balance of Bcl-2/Bax, which would trigger caspases-induced apoptosis. These data provide preclinical support for chemotherapeutic approach with matrine for the treatment of gallbladder cancer. All these results indicate that matrine may represent a novel class of chemotherapeutic agents for gallbladder cancer.