ABSTRACT
OBJECTIVE：To study the effects of costunolide （COS）on the cell cycle distribution and apoptosis of human breast cancer SK-BR- 3 cells and its mechanism. METHODS ：Human breast cancer SK-BR- 3 cells were divided into blank control group，and COS groups of 10，20，30，40，50 μmol/L. MTT assay was used to detect the effects of COS on cell proliferation. SK-BR-3 cells were divided into blank control group ，COS low ，medium and high concentration groups （10，20，30 μmol/L）. After cultured for 24 h，flow cytometry was used to detect the distribution of cell cycle. Hoechst 33258 fluorescence staining was used to detect cell apoptosis. Western blot assay was used to detect the expression of p 53，caspase-3，Bcl-2，Bax，p21，CDK2 and cyclinE. RESULTS ：Compared with blank control group ，COS could significantly inhibit the proliferation of SK-BR- 3 cells（P＜ 0.05 or P＜0.01），and in a dose and time-dependent manner. Low ，medium and high concentrations of COS could induce cell apoptosis and arrest cell at G 1/S phase （P＜0.05 or P＜0.01），could significantly up-regulate the protein expression of p 53， caspase-3，Bax and p 21（P＜0.05 or P＜0.01），and could significantly down-regulate the protein expression of Bcl- 2，CDK2 and cyclinE（P＜0.01）. CONCLUSIONS ：COS can inhibit the proliferation of human breast cancer SK-BR- 3 cells and induce cell apoptosis and cell cycle arrest. The mechanism may be related to the regulation of p 53/Bax/Bcl-2/caspase-3 apoptosis signal pathway.
ABSTRACT
OBJECTIVE：To study the effects of costunolide on the proliferation ，migration and apoptosis of breast cancer SK-BR-3 cells and its mechanism. METHODS ：SK-BR-3 cells in logarithmic growth period were collected and cultured with different concentrations （10，20，30，40，50 μmol/L）of costunolide for 24，48，72 h. Inhibitory rate of costunolide on cell proliferation was detected with CCK- 8. The cells were divided into blank control group and costunolide group （10，20，30 μmol/L）. Hoechst 33258 fluorescence was used to observe the morphology and apoptosis of cells ，and apoptotic rate of cells were calculated. Cell scratch test was used to detect the migration ability of cells and calculate the migration rate. Western blotting was used to detect the relative expression level of Bcl- 2，Bax，Caspase-3 and Cleaved Caspase- 3 in cells. RESULTS ：The proliferation of SK-BR-3 cells were significantly inhibited by costunolide （P＜0.05 or P＜0.01），and it shows a trend of concentration and time dependence. In the blank control group ，cells possessed clear contour ，regular shape and good adherence . Compared with blank control group，the number of cells were decreased significantly in 10，20，30 μmol/L costunolide groups，the cell structure was loose，the volume was reduced ，and the gap became larger ，and most of the cell contour disappeared and became round ，the cell adherence was poor ；cell migration rate and Bcl- 2 protein relative expression level were decreased significantly ，while apoptosis rate and the relative expression level of Bax ，Caspase-3 and Cleaved Caspase- 3 protein were significantly increased （P＜0.05 or P＜0.01）. CONCLUSIONS ： Costunolide can inhibit the proliferation and migration ，and induce apoptosis of human breast cancer SK-BR- 3 cells，mechanism of which may be through up-regulating the expression of Bax ，Caspase-3 and Cleaved Caspase- 3 while down-regulating the expression ofBcl-2.
ABSTRACT
Exosomes are vesicles of endocytic origin released from different cell types under both normal and pathological conditions.Proteomics is an emerging discipline for studying composition and mechanics and interac-tion of proteins with regard to disease occurrence、cellular metabolism,etc.Progress of research on exosomes from tumor cell at proteomics technology platforms in aspect of clarifying mechanisms of shaping and sustaining micro-enviroment of tumor growth and survival and metastasis, induce tumor proliferation, Immunosuppressive effects or immune evasion, and discovering biomarker, makes a promising prospect of exsomes in clinical diagnosis and treatment.